• Title/Summary/Keyword: soft agar

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Strain Improvement by Interspecific Fusion of Streptomyces actuosus DMCJ-49 and Streptomyces minoensis DMCJ-144 producing $\alpha$-Amylase Inhibitor ($\alpha$-Amylase 저해제 생성균 Streptomyces actuosus DMCJ-49와 Streptomyces minoensis DMCJ-144의 종간 융합에 의한 균주 개량)

  • 김지현;최응칠;김병각
    • YAKHAK HOEJI
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    • v.35 no.1
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    • pp.30-37
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    • 1991
  • Streptomyces actuosus DMCJ-49 and Streptomyces minoensis DMCJ-144 produce the .alpha.-amylase inhibitor. Inerspecific protoplast fusion technique was used to increase the productivity of .alpha.-amylase inhibitor. Four auxotrophic mutants were obtained respectively from two strains by N-methyl-N'-nitor-N-nitrosoguanidine(3mg/ml) treatment. The optimum conditions for the protoplast formation of Streptomyces actuosus DMCJ-49 ade was as follows; 1.2% w/v of glycine, 3mg/ml of lysozyme, and 30 min of lysozyme treatment followed by 36 hr. incubation in the protop-last formation medium. In case of DMCJ-144-his those were 1.2%w/v, 3 mg/ml, 30 minutes and 60 hours, respectively. Regeneration was accomplished with hypertonic soft agar medium that contained 0.4M sucrose, 20mM CaCl$_2$, 50 mM MgCl$_2$ and low levels of phosphate. Fusion of protoplasts carrying different auxotrophic markers was achieved by treatment with polyethylene glycol. The optimum concentration of polyethylene glycol 1450 for the production of recombinants was 40%w/v. When the protoplasts was treated with 40% polyethylene glycol for 30 minutes, the frequency of recombinants was 6.5$\times$$10^{-3}$ and the $\alpha$-amylase inhibition activity of $ade^-his^-$ No. 4, which is the fusant with the most improved activity increased from 33 to 125 I.U./ml.

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Antimicrobial Effect of Low Temperature Atmospheric Plasma against Oral Pathogens

  • Kim, Young Min;Choi, Byul Bo Ra;Park, Sang Rye;Kim, Ji Young;Kim, Gyoo Cheon
    • International Journal of Oral Biology
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    • v.40 no.4
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    • pp.167-173
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    • 2015
  • The purpose of this study was to investigate the antibacterial effect of the low temperature atmospheric plasma device with needle tip designed for easy approach to the oral cavity and root canal against Streptococcus mutans, Enterococcus faecalis and Candida albicans. The antibacterial activities evaluated by measuring clear zone of agar plate smeared with each bacteria after plasma treatment. To quantify antibacterial effects, dilution plate method was used. In addition, scanning electron microscope (SEM) was used for observation of changes in bacterial morphology. As treatment time of plasma increased, the clear zone was enlarged. The death rate was more than 99%. The SEM results showed that the globular shape of bacteria was distorted. These results suggest that needle tip plasma could be an innovative device for prevention of dental caries, and treatment of apical infection and soft tissue diseases.

Novel p104 protein regulates cell proliferation through PI3K inhibition and p27Kip1 expression

  • Han, Seung-Jin;Lee, Jung-Hyun;Choi, Ki-Young;Hong, Seung-Hwan
    • BMB Reports
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    • v.43 no.3
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    • pp.199-204
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    • 2010
  • The protein p104 was first isolated as a binding partner of the Src homology domain of phospholipase C$\gamma$1, and has been shown to associate with p85$\alpha$, Grb2. The ectopic expression of p104 reduced cellular growth rate, which was also achieved with the overexpression of only the proline-rich region of p104. The proline-rich region of p104 has been found to inhibit the colony formation of platelet-derived growth factor BB-stimulated NIH3T3 cells and MCF7 cancer cells on soft agar. Mutagenesis analysis showed that the second and third proline-rich regions are essential for growth control, as well as for interaction with p85$\alpha$. Overexpression of p104 increased the level of the cyclin-dependent kinase inhibitor, $p27^{Kip1}$, and inhibited the activity of phosphoinositide 3-kinase (PI3K). In summary, p104 interacts with p85$\alpha$ and is involved in the regulation of $p27^{Kip1}$ expression for the reduction of cellular proliferation.

Characteristics and application of monoclonal antibody to progesterone I. Production of monoclonal antibody to progesterone (Progesterone의 단크론성 항체에 관한 특성 및 활용에 관한 연구 I. 단크론성 항체의 생산)

  • Kang, Chung-boo;Kim, Yong-hwan
    • Korean Journal of Veterinary Research
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    • v.30 no.4
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    • pp.511-513
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    • 1990
  • Monoclonal antibody to progesterone was produced using the antigen $11{\alpha}$-hydroxyprogesterone hemisuccinate conjugated to bovine serum albumin. Hybridomas secreting antibody to progesterone were detected by radioimmunoassay and enzyme-linked immunosorbent assay and cloned in soft agar. Two stable monoclonal antibodies which were highly specific to progesterone were obtained, so it may be advantageously used to study on several physiological functions of progesterone including immunological research.

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Diarylbutane-type Lignans from Myristica fragrans (Nutmeg) show the Cytotoxicity against Breast Cancer Cells through Activation of AMP-activated Protein Kinase

  • Le, Thi Van Thu;Nguyen, Phi Hung;Choi, Hong Seok;Yang, Jun-Li;Kang, Keon Wook;Ahn, Sang-Gun;Oh, Won Keun
    • Natural Product Sciences
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    • v.23 no.1
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    • pp.21-28
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    • 2017
  • In our program to search for new AMP-activated protein kinase (AMPK) activators from plants that exert potential anticancer property, we found that an EtOAc extract of Myristica fragrans (nutmeg) activated AMPK enzyme in human breast cancer MCF-7 cells. Two major diarylbutane-type lignans, macelignan and meso-dihydroguaiaretic acid (MDGA), were isolated as active principles from this extract. Treatment of breast cancer cells with two compounds induced cellular apoptosis, evidenced by cleavage of poly-(ADP-ribose) polymerase (PARP) and Ser 15 phosphorylation of p53. Moreover, macelignan and MDGA significantly inhibited the colony formation of MCF-7 breast cancer cells on soft agar. Intraperitoneal injection of macelignan and MDGA (20 mg/kg) suppressed the tumor growth of 4T1 mammary cancer cells. These results indicate that the chemopreventive effects of two major diarylbutane-type lignans from Myristica fragrans (nutmeg) may be associated with induction of apoptosis presumably through AMPK activation.

Plant Regeneration from Protoplasts Isolated through Embryogenic Cell suspension Culture in Rice (벼 현탁배양을 통하여 분리된 원형질체로부터 식물체 재분화)

  • 정병균
    • Journal of Plant Biology
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    • v.36 no.3
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    • pp.211-218
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    • 1993
  • Plant regeneration was accomplished from protoplast culture of rice (Oryza sativa L. cv. Taebaeg). Embryogenic callus was induced from mature seed on MS medium containing 5 mM proline, 2.5 mg/L 2,4-D, 30 g/L sucrose in the dark at 28$^{\circ}C$ and used to establish embryogenic cell suspension culture. Suspension cells were subcultured every one week in N6 medium supplemented with 5 mM proline, 200 mg/L casein hydrolysate, 2.5 mg/L 2,4-D and amino acids of AA medium. Suspension cultures were composed of cells that were densely cytoplasmic, potentially embryogenic and were at least maintained for more than 6 months in liquid medium. Protoplasts were isolated from fast-growing suspension culture cells and cultured in a slightly modified KpR medium by mixed nurse culture. Isolated protoplasts began to divide within 5~7 days and thereafter, protoplast-derived calli were sequentially transferred to callus proliferating medium that soft agar MS medium contained 2 mg/L 2,4-D and produced distinct embryogenic cells. Microcolonies were then transferred to solid medium which consisted of MS medium containing 5 mg/L kinetin, 1 mg/L NAA, 1 mg/L ABA, 30 g/L sucrose and 10 g/L sorbitol under fluorescent light. Mulitple shoots of 4~5 per callus emerged and were transferred to hormone-free MS medium for root initiation. Thereafter, The plantlets were transferred to pots of soil to mature in the culture room.

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Carcinogenic Potentials of HPV-16 and NNK in Human in Vitro Model (인체 세포 모델을 이용한 HPV-16과 NNK의 발암 잠재력에 관한 연구)

  • 양재호;이세영
    • Toxicological Research
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    • v.12 no.2
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    • pp.271-275
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    • 1996
  • Carcinogenic potential of HPV-16 DNA and NNK in a human keratinocyte cell line was assessed to study effects of viral-chemical interaction. Human cells were transfected with HPV-16 DNA and 6 clonal cell lines were subsequently obtained. Clonal line-3 and 6 at passage 7 showed characteristics of tumor cells such as increases of saturation density, soft-agar colony formation, cell aggregation and foci appearance. Among cells treated with 1$\mu M$, 10$\mu M$, 100$\mu M$ or 1 mM of NNK for 4 weeks, 100$\mu M$ treatment showed most tumorigenic characteristics at passage 7. These results indicate that either HPV-16 or NNK alone is tumorigenic in this in human in vitro model. When cells transfected with HPV-16 were subsequently exposed by 100 uM NNK for 4 weeks, all the clonal cells except clone-1 showed higher levels of tumor cell characteristics than HPV-16 DNA or NNK exposure alone. Clonal line-6, the most tumorigenic cells, showed higher transcriptional level of fibronectin and lower level of TGF-$\beta_1$, as compared to control cells, suggesting that alteration of growth factor or extracellular matrix may play a role in carcinogenesis process induced by HPV-16 and NNK. Taken together, the present study indicates that viral-chemical interactions between HPV-16 DNA and NNK enhance carcinogenic potentials of human cells and implies that smoking among people infected with human papillomavirus may pose an additional risk of causing cancer.

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MiR-371 promotes proliferation and metastasis in hepatocellular carcinoma by targeting PTEN

  • Wang, Hao;Zhao, Yi;Chen, Tingsong;Liu, Guofang;He, Nan;Hu, Heping
    • BMB Reports
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    • v.52 no.5
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    • pp.312-317
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    • 2019
  • Hepatocellular carcinoma (HCC) is the leading cause of cancer-related mortality worldwide. MiR-371 has recently emerged as an important regulator in tumorigenesis, and may serve as a biomarker for malignant tumors. We transfected miR-371 or its inhibitor in two human HCC cell lines, then used 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, soft agar colony formation, and transwell migration assays to evaluate the effects on cell proliferation, migration, and invasion. We found that miR-371 was positively correlated with HCC metastasis and poor prognosis in the inflicted patients, and the high expression of miR-371 was promoted, whereas a low level of miR-371 depressed cell proliferation and invasion. We found PTEN to be a direct target of miR-371. The overexpression or knockdown of PTEN exhibited the opposite effects from those of miR-371 on cell proliferation and migration. Our study demonstrates that miR-371 promotes proliferation and metastasis in HCC by targeting PTEN.

Occurrence of Strawberry Soft Rot by Rhizopus stolonifer on Distribution after Harvest (수확후 유통중 Rhizopus stolonifer에 의한 딸기 무름병 발생)

  • Kwon, Jin-Hyeuk;Yoon, Hae-Suk;Nam, Myeong-Hyeon;Park, Chang-Seuk
    • The Korean Journal of Mycology
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    • v.37 no.1
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    • pp.117-119
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    • 2009
  • Rhizopus soft rot caused by Rhizopus stolonifer occurred on strawberry (Fragaria ${\times}$ ananassa) in the plastic boxes in Jinju City Wholesale Market of Agricultural Products at Gyeongnam province in Korea on April 2008. The infected fruits were rapidly softened, water soaked and finally rotted. The symptoms were mainly occurred through wounds at harvest time. The colony color was white, cottony to brownish black after fully matured and numerous sporangiospores were formed on sporangia. The optimum growth temperature of the fungus was $30^{\circ}C$ on potato dextrose agar. Sporangia were globose or hemispheric shape of white color at initial stage and gradually changed with black color of $82{\sim}195{\mu}m$ in size. Sporangiophores were $12{\sim}25{\mu}m$ in width. Columella were hemispheric in shape, and $70{\sim}92{\mu}m$ in size. Sporangiospores were irregular round or oval in shape, brownish-black in color and $9{\sim}21\;{\times}\;7{\sim}8{\mu}m$ in size. This fungus formed stolones and rhizoid on PDA. On the basis of symptom, mycological characteristics and pathogenicity of the fungus, the causal fungus was identified as Rhizopus stolonifer (Ehrenb.) Vuill.

Quality and Sensory Characteristics of Yanggaeng, a Healthy Snack Added with Pinus Koraiensis Needle Extract Powder for the Elderly

  • Eunbin Park;Soo In Ryu;Minho Lee;Hyo-Jeong Lee;Jean Kyung Paik
    • Journal of Food and Nutrition Research
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    • v.10 no.3
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    • pp.228-234
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    • 2022
  • Korea has entered the aged society, with those aged over 65 years accounting for 14~20% of its population. Interest in the quality of life, nutrition, and health of the elderly is increasing. Since the energy intake of the elderly is lower than the estimated required amount, nutrient intake ratio of the elderly is related to mastication. Yanggaeng is a high-energy food made from agar, sugar, and red bean paste. Since it has a soft texture, it is highly utilized as a snack for the elderly who have inconvenience in mastication. Pinus koraiensis needle is known to possess antioxidant and antibacterial effects. The purpose of this study was to develop a functional snack added with Pinus koraiensis needle extract powder known to possess anti-inflammatory, anti-cancer, and antibacterial effects for the elderly that would be easy for them to chew. In this study, yanggaeng was manufactured with 0%, 2%, 4%, 6%, or 8% of Pinus koraiensis needle extract powder, white sediment, and agar. In the experiment, moisture contents, color values texture characteristics, antioxidant activities, and sensory preference results were evaluated. Moisture content was the highest in the group added 6% of the extract powder. It was the lowest in the group added 8% of the extract powder, showing a significant (p < 0.001) difference between the two. Hardness, cohesiveness, and springiness of texture characteristics were decreased significantly (p < 0.001) while the adhesiveness and chewiness of texture characteristics were increased significantly (p < 0.001) as the amount of addition increased. Regarding antioxidative activity measurements, levels of polyphenols were the highest (p < 0.001) in the group added with 8% of the extract powder. DPPH and ABTS radical scavenging activities were increased as the amount of addition increased (p < 0.001). Overall sensory preference was the highest for the 2% addition group. It decreased as the amount of addition increased (p < 0.001). Results of this study indicate that Pinus koraiensis needle extract powder with antioxidant and antibacterial effects can be used to manufacture yanggaeng to make functional snacks with improved quality characteristics for the elderly.