• Title/Summary/Keyword: Growth inhibition test

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Cloning, expression, and activity of type IV antifreeze protein from cultured subtropical olive flounder (Paralichthys olivaceus)

  • Lee, Jong Kyu;Kim, Hak Jun
    • Fisheries and Aquatic Sciences
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    • v.19 no.8
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    • pp.33.1-33.7
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    • 2016
  • Antifreeze proteins (AFPs) lower the freezing point but not the melting point of aqueous solutions by inhibiting the growth of ice crystals via an adsorption-inhibition mechanism. However, the function of type IV AFP (AFP IV) is questionable, as its antifreeze activity is on the verge of detectable limits, its physiological concentration in adult fish blood is too low to function as a biological antifreeze, and its homologues are present even in fish from tropic oceans as well as freshwater. Therefore, we speculated that AFP IV may have gained antifreeze activity not by selective pressure but by chance. To test this hypothesis, we cloned, expressed, and assayed AFP IV from cultured subtropical olive flounder (Paralichthys olivaceus), which do not require antifreeze protein for survival. Among the identified expressed sequence tags of the flounder liver sample, a 5'-deleted complementary DNA (cDNA) sequence similar to the afp4 gene of the longhorn sculpin was identified, and its full-length cDNA and genome structure were examined. The deduced amino acid sequence of flounder AFP IV shared 55, 53, 52, and 49 % identity with those of Pleuragramma antarcticum, Myoxocephalus octodecemspinosus, Myoxocephalus scorpius, and Notothenia coriiceps, respectively. Furthermore, the genomic structure of this gene was conserved with those of other known AFP IVs. Notably, the recombinant AFP IV showed a weak but distinct thermal hysteresis of $0.07{\pm}0.01^{\circ}C$ at the concentration of 0.5 mg/mL, and ice crystals in an AFP IV solution grew star-shaped, which are very similar to those obtained from other polar AFP IVs. Taken together, our results do not support the hypothesis of evolution of AFP IV by selective pressure, suggesting that the antifreeze activity of AFP IV may have been gained by chance.

Development of Enterococcus faecalis Strains Resistant to Rifampicin and Ofloxacin (리팜피신과 오플로삭신에 내성인 Enterococcus faecalis 균주의 개발)

  • Lee, Soo-Hwa;Kim, Sook-Kyung;Chung, Young-Ja;Shim, Mi-Ja;Kim, Byong-Kak;Choi, Eung-Chil
    • YAKHAK HOEJI
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    • v.40 no.3
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    • pp.351-356
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    • 1996
  • The preparation of Enterococcus faecalis RSI is used as a therapeutics for human intestinal disorders. However, the microbe in this preparation is usually very sensitive to rifampicin and fluoroquinolones. If this preparation is taken with rifampicin or fluoroquinolones, its therapeutic effect can not be expected. E. faecalis RFR11, containing resistance to rifampicin was obtained by MNNG mutation method. Serial passage of E. faecalis RFR11 produced E. feacalis OFR16 on agar with 2-fold minimal inhibitory concentration of ofloxacin produced. E. feacalis OFR16 was resistant to fluoroquinolones up to 8-256 fold higher than that for the original strain. E. faecalis OFR16 also exhibited identical characteristics with the parent strain when they were tested for lactic acid formation and growth inhibition of E. coli MB4-5737 and Shigella sonnei MB4-10411. From in vitro test, it was identified that rifampicin and ofloxacin is not inactivated by certain factors of E. faecalis OFR16. Conclusively. E. faecalis OFR16, rifampicin and fluoroquinolones resistant mutant, is an efficient strain that has insensitivity against rifampicin and fluoroquinolones and original biochemical characteristics of the parent strain.

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The Study of Antibiotic Resistance in Bacterial Biofilms (박테리아 생체막에 대한 항생제 내성 연구)

  • Kim Jin Wook;Joo Chi Un;Park Jin Yong;Lee Song Ae;Kim In Hae;Lee Jae Hwa
    • Environmental Mutagens and Carcinogens
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    • v.25 no.4
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    • pp.157-160
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    • 2005
  • Antibiotic resistance of bacteria in the biofilm mode of growth contributes to the chronicity of infection and disease. The penetration of antibiotic, through biofilm developed in an itt vitro model system was investigated. Antibiotic resistant bacteria (E. coli) were obtained from Culture Collection of Antibiotic Resistant Microbes. Ca-alginate bead used as simulated biofilm and a cell entrapment test using compressed air were experiment for the improvement cell viability. Antibiotic susceptibilities though biofilms was measured by assaying the concentration of antibiotic that diffused through the biofilm to minimal inhibition concentration (MIC). Survival of immobilized cells were reduced as compared to free cells. In case of antibiotic susceptible E. coli reduced continuously, but antibiotic resistant E. coli kept up survival rate constantly. Survival was showed after exposed to the antibiotics that the more treated antibiotic resistant E. coli and low concentration of antibiotics) the more survived.

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Effects of Environmental Factors on Nitrite Accumulation in a Strong Nitrogen Removal System (고농도 질소폐수 처리 공정에서 환경인자가 아질산염 축적에 미치는 영향)

  • Park, Noh-Back;Choi, Woo-Yung;Yoon, Ae-Hwa;Jun, Hang-Bae;Park, Sang-Min
    • Journal of Korean Society of Water and Wastewater
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    • v.24 no.1
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    • pp.51-62
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    • 2010
  • The high concentration of N in the wastewater from livestock farming generally renders the efficiency of the wastewater treatment. Therefore, removal of N in livestock wastewater is crucial for successful treatment. The current study was conducted to investigate the optimum conditions for partial nitrification under anaerobic condition following nitritation in TPAD-BNR(two-phase anaerobic digestion-biological nitrogen removal) operating system. Sequential operating test to stimulate partial nitrification in reactor showed that partial nitrification occurred at a ratio of 1.24 in $NO_2{^-}$-N:$NH_4{^+}$-N. With this result, a wide range of factors affecting stable nitritation were examined through regression analysis. In the livestock wastewater treatment procedure, the hydraulic retention time (HRT) and pH range for optimum nitrite accumulation in the reactor were 1-1.5 days and 7-8, respectively. It was appeared that accumulation of $NO_2{^-}$-N in the reactor is due to inhibition of the $NO_2{^-}$-N oxidizer by free ammonia (FA) while the effect of free nitrous acid was minimal. Nitrification was not influenced by DO concentration at a range of 2.0-3.0 mg/L and the difference in the growth rate between $NH_4{^+}$-N oxidizer and $NO_2{^-}$-N oxidizer was dependent on the temperature in the reactor.

Chemical Components, Antitermite and Antifungal Activities of Cinnamomum parthenoxylon Wood Vinegar

  • ADFA, Morina;ROMAYASA, Ari;KUSNANDA, Arif Juliari;AVIDLYANDI, Avidlyandi;YUDHA S., Salprima;BANON, Charles;GUSTIAN, Irfan
    • Journal of the Korean Wood Science and Technology
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    • v.48 no.1
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    • pp.107-116
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    • 2020
  • Termiticidal and fungicidal activities of wood vinegar from Cinnamomum parthenoxylon (CP) stem wood have been evaluated against Coptotermes curvignathus and wood rotting fungi (Schizophyllum commune and Fomitopsis palustris). The utilized CP wood vinegar was produced in the operating temperature range 250-300℃ pyrolysis. A no-choice test was applied for evaluating termiticidal activity with 33 active termites and antifungal activity using the agar media assay. The result showed that an increase in the concentrations of CP wood vinegar significantly raised the mortality of termite. CP wood vinegar showed high termiticidal activity, organic acids (acetic acid 42.91%, 3-butenoic acid 6.89%, butanoic acid, 2-propenyl ester 2.26%), and ketones (1-hydroxy-2-propanone 5.14%, 3-methylcyclopentane-1,2-dione 2.34%) might be largely contributed to termiticidal activity in addition to other minor components. Furthermore, CP wood vinegar exhibited significant inhibition of fungal growth. These data showed that CP wood vinegar was more toxic to white-rot fungi (S. commune) than brown-rot (F. palustris). The results suggested that phenolic compounds from lignin degradation were responsible for good antifungal activity.

Anti-tumorigenic and Invasive Activity of Colon Cancer Cells Transfected with the Retroviral Vector Encoding Tissue Inhibitor of Metalloproteinase-2 (레트로바이러스를 이용한 Tissue Inhibitor of Metalloproteinase-2 유전자 발현이 대장암 세포의 전이 및 종양형성에 미치는 영향)

  • 오일웅;정자영;장석기;이수해;김연수;손여원
    • YAKHAK HOEJI
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    • v.48 no.3
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    • pp.189-196
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    • 2004
  • Matrix metalloproteinases (MMPs) and tissue inhibitors of MMPs (TIMPs) playa key role in tumor invasion and metastasis. As an inhibitor of MMP-2, TIMP-2 is known to block both the invasive and metastatic behavior of cancer cells, and decrease tumor growth activity. We performed this study to investigate the effects of TIMP-2 over-expression induced by retroviral mediated gene transfer in vitro and in vivo. The human colon cancer cell line SW480 was transfected with the retroviral vector encoding TIMP-2. The effects of TIMP-2 over-expression were analyzed by invasion assay and gelatinase activity test in colon cancer cells and tumorigencity in nude mice. In evaluation of the transfection efficiency of the retroviral vector encoding TIMP-2 in colon cancer cells, we confirmed up-regulation of TIMP-2 expression dependent on the time of cell culture. In addition, inhibition of MMP-2 expression in SW480/TIMP-2 was shown by gelatin zymography. In the in vitro invasion assay SW480/TIMP-2 inhibited the invasiveness on matrigel coated with collagen. To determine whether TIMP-2 can modulate in vivo tumorigenicity and metastasis, SW480/TIMP-2 cells were injected subcutaneously in nude mice. The tumor mass formation of SW480/TIMP-2 cells in nude mice was markedly decreased compared to nontransfected cancer cells. These results showed that colon cancer cells transfected with the retroviral vector encoding TIMP-2 inhibits the invasiveness in vitro and tumorigenicity in vivo.

Bacillus coagulans OFR17 Strain Resistant to Rifampicin and Ofloxacin (리팜피신과 오플록사신에 내성인 Bacillus coagulans OFR17 균주)

  • Kim, Eun-Ah;Oh, Tae-Kwon;Choi, Keum-Hwa;Lee, Jin-Hee;Baek, Moon-Chang;Kim, Byong-Kak;Choi, Eung-Chil
    • YAKHAK HOEJI
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    • v.41 no.4
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    • pp.450-455
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    • 1997
  • The preparation of Bacillus coagulans is used as a therapeutics for human intestinal disorders. However, the bacterium in the preparation is very susceptible to rifampic in and fluoroquinolones. When the preparation is taken with rifampicin or fluoroquinolones, its therapeutic effect can not be expected. So B. coagulans RFR17 resistant to rifampicin was obtained by treating the parent B. coagulans with N-methyl-N'-nitro-N-nitrosoguanidine. B. coagulans OFR17 was produced by serial passage of B. coagulans RFR17 on agar with 2-fold minimal inhibitory concentration of ofloxacin or ciprofloxacin. B. coagulans OFR17 was resistant to fluoroquinolones up to 16~64 fold higher than that for the original strain. B. coagulans OFR17 also exhibited identical characteristics with the parent strain when they were tested for lactic acid production and growth inhibition of E. coli MB4-01 and Shigella sonnei MB4-10411. From in vitro test, it was also identified that rifampicin and ofloxacin are not inactivated by certain factors of B. coagulans OFR17. Conclusively, B. coagulans OFR17 can be regarded as a promising strain which can be developed as the preparation for the treatment of the intestinal disorders of the tuberculosis patients under rifampicin and ofloxacin therapy.

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Biological control of powdery mildew in Korean melons through a treatment with a culture of Bacillus species

  • Lee, Sang Gil;Jeon, Nak Beom;Park, Myung Soo;Yun, Hae Keun
    • Korean Journal of Agricultural Science
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    • v.47 no.4
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    • pp.1011-1020
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    • 2020
  • Efficient management systems of facilities make it possible to manage environmental conditions properly, such as the temperature, humidity and light source required for the best growth of the crops, as well as for the mass production of fruit and vegetables with high quality every year through an advanced and protected cultivation system. Powdery mildew is a type of chronic disease that is difficult to control during the production of Korean melons under a protected cultivation system, the use of which is increasing in production areas in Korea. Two Bacillus strains isolated from soil samples showed antagonistic activities against several pathogens, specifically Botrytis cinerea, Colletotrichum gloeosporioides, and Fusarium oxysporum f.sp. melonis; they were identified as Bacillus velezensis M2 and B. amyloliquefaciens M3 in a molecular biological test of the nucleotide sequences of gyrase subunit A (gyrA). The treatment was given three times at intervals of five days with 400-fold diluted cultures of B. velezensis M2 and B. amyloliquefaciens M3. This led to the inhibition of the incidence of powdery mildew disease in Korean melon leaves, which resulted in effective control efficiency against the incidence of powdery mildew disease with control values of 87% and 65%, respectively. Cultures of antagonistic microbes tested in this study can be used to increase the efficiency as part of an environmentally friendly management scheme to prevent powdery mildew disease during the protected cultivation of crops, including Korean melons.

Response to Fungicides of Colletotrichum spp. Isolated from Red Peppers in Sunchang, Korea (순창군 지역의 고추로부터 분리한 탄저병균의 살균제에 대한 반응)

  • Ju, Hyeon-Yeong;Son, Mi-Ra;Kim, Jin-Won
    • Research in Plant Disease
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    • v.27 no.4
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    • pp.164-171
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    • 2021
  • In August 2020, 377 anthracnose strains were isolated from anthracnose-infected peppers collected from 25 farms in Sunchang-gun, Jeollabuk-do. Inhibition rate of mycelial growth of 11 pepper anthracnose fungicides registered in Korea was investigated for 62 strains selected by region and the degree of susceptibility to each fungicide was investigated. As a result of the fungicide susceptibility test of anthracnose to the fungicide, no resistant strains were observed in fluazinam, prochloraz manganese, and benomyl, but resistant strains appeared in at least three areas for other fungicides, and all strains in all regions were resistant to dithianon. Through this, it was confirmed that the fungicide resistance was expressed in the strain group due to the continuous treatment of the fungicide in some areas. By region, resistant strains to seven pesticides appeared in Sunchang-eup and Paldeok-myeon, and resistant strains to six pesticides appeared in Geumgwa-myeon, Bokheung-myeon, Ssangchi-myeon, Yudeung-myeon, and Pungsan-myeon. There was no significant correlation between fungicide registration time and emergence of resistant strains.

Studies on Biological Activity of Wood Extractives (X) - Antifungal Compounds of Hovenia dulcis - (수목추출물의 생리활성에 관한 연구(X) - 헛개나무 목부의 항균활성 물질 -)

  • Choi, Yun-Jeong;Lee, Hak-Ju;Lee, Sung-Suk;Choi, Don-Ha
    • Journal of the Korean Wood Science and Technology
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    • v.31 no.1
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    • pp.1-9
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    • 2003
  • Antimicrobial activities of plant extractives were investigated to develop a natural fungicide. Two stilbenoids and five flavonoids were isolated from wood extractives of Hovenia dulcis (Rhamnaceae) which had been selected due to its high antifungal activity among the tested species. The chemical structures of isolated compounds were determinded as : 5-hydroxy-7-methoxyflavone, 5,7-dihydroxyflavone (chrysin), 5,7-dihydroxyflavanone (pinocembrin), 3,5,7-trihydroxyflavanone (pinobanksin), 3,4',5,7-tetrahydroxyflavanone (aromadendrin), 3-hydroxy-5-methoxystilbene and 3,5-dihydroxystilbene (pinosylvin) on the basis of Mass and NMR spectroscopic data. According to the results of antifungal test, 3-hydroxy-5-methoxystilbene was evaluated as the strongest antifungal compound among the tested compounds and next were pinocembrin and pinosylvin, but those also had high hyphal growth inhibition activities against C. parasitica, T. versicolor, T. palustris and T. viride. However, pinobanksin, 5-hydroxy-7-methoxyflavone, chrysin and aromadendrin showed very low antifungal activity. In this regard, it could inferred that high antifungal activity of wood extractives of H. dulcis were derived from 3-hydroxy-5-methoxystilbene, pinocembrin and pinosylvin, respectively.