• Journal of Environmental Science International
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• v.20 no.9
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• pp.1087-1093
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• 2011

The mixotrophic marine ciliate Mesodinium rubrum possesses a highly modified algal endosymbiont as a nutrition source for the species. Accordingly, we assumed that the species can reflect the ecotoxicity on marine producer (as phytoplankton) and consumer (as zooplankton) both. A series of experiments were conducted to identify the potential of the species as a standard test species for marine ecotoxicological study. The comparison of species sensitivity on reference toxic materials was made using potassium dichromate for phytoplankton and copper chloride for zooplankton. The ciliate revealed the highest sensitivity on both reference materials among the seven test species including phytoplankton, benthic copepod and rotifer species. The toxicity end point of the species was 72hr-$EC_{50}$=1.52 mg/L (as potassium dichromate) estimated by population growth inhibition (PGI), which is more sensitive than the most sensitive phytoplankton Skeletonema costatum (72hr-$EC_{50}$=3.05 mg/L). As comparison to rotifer, it also revealed higher sensitivity on copper chloride; 72hr-$EC_{50}$=0.38 mg/L for ciliate and 48hr-$EC_{50}$=0.48 mg/L for rotifer. Also, the elutriate toxicity test of various ocean disposal wastes were conducted to identify the potential of ciliate toxicity test application using industrial waste sludges. The toxicity of leather processing waste sludge was highest on the ciliate, followed by dyeing waste sludge and dye production waste sludge as an increasing order of toxicity. 72h-$EC_{50}$ of ciliate PGI test was 1.83% and that of S. costatum 3.84% for leather waste sludge which showed highest toxicity. The toxicity test results also revealed that the highest sensitivity was observed on ciliate species on ocean disposed sludge wastes. Also, ciliate toxicity test well discriminated the degree of toxicity between sludge sources; 72h-$EC_{50}$ values were 1.83% for leather processing waste sludge, 16.75% for dye production waste sludge and 27.75% for textile production waste sludge. Even the laboratory culture methods of the species were not generally established yet, the species has high potential as the standard test species for marine toxicity test in terms of the dual reflection of phyto- and zooplankton toxicity from single test, sensitivity and test replicability.

• Research in Plant Disease
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• v.12 no.3
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• pp.260-266
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• 2006

Pythium blight caused by Pythium spp. is one of major diseases in putting green of golf course. In this study, microorganisms which are anatgonistic to Pythium aphanidermatum, a pathogen of pythium blight, were selected primary through in vitro tests, dual culture method and triple layer agar diffusion method. In vivo test against pythium blight were conducted to select the best candidate biocontrol microorganism by pot experiment in a plastic house. Bacillus subtilis GB-0365 was finally selected as a biocontrol agent against pythium blight. Relative Performance Indies(RPI) was used as a criterion of selecting potential biocontrol agent. B. subtilis GB-0365 showed resistance to major synthetic agrochemicals used in golf course. Alternative application of synthetic agrochemicals and B. subtilis GB-0365 was most effective to successfully contol pythium blight. B. subtilis GB-0365 suppressed the development of pythium bight of bentgrass by 56.4% as compared to non-treated control and its disease control efficacy was 60.9% of a synthetic fungicide Oxapro(WP) efficacy. B. subtilis GB-0365 has a potential to be a biocontrol agent for control of pythium blight.

• Korean Journal Plant Pathology
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• v.3 no.3
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• pp.187-197
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• 1987

Bacteria and fungi antagonistic to Fusarium oxysporum f. sp. cucumerinum Owen were effectively isolated with each of modified Triple Layer Agar (TLA) technique from rhizosphere soil where cucumber had been grown healthily in plastic film house. Three predominant bacterial isolates selected were identified as Pseudomonas fluorescens, and P. putida, Serratia sp. and three fungal isolates were Gliocladium sp. Trichoderma harzianum, and T. viride. Antagonistic bacteria inhibited $26-45\%$ of germination and $41-56\%$ of germ tube elongation of microconidia of F. oxysporum f. sp. cucumerinum on Water Agar (WA). P. fluorescens was the strongest inhibitor. Several my co parasitisms were observed on dual culture of WA between antagonistic fungi and F. oxysporum f. sp. cucumerinum such as coiling, penetration, overgrowing, and lysis. Mycelial lysis of the pathogen was the most severe at pH 4.6, followed by 3.6, 5.6 and 6.6 of the medium in decreasing order. At pH 6.6, mycelia of the pathogen were not conspicuously damaged, however, the antagonistic fungi formed abundant chlamydospores especially Gliocladium sp. T. harzianum revealed the most excellent antagonism in vitro.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.10
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• pp.5825-5831
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• 2013

Background/Objectives: Maintenance of cellular function in culture is vital for transfer and development following adoptive immunotherapy. Dual properties of IL-21 in activating T cells and reducing activation induced cell death led us to explore the mechanism of action of IL-21 enhanced proliferation and cytotoxic potential of CIK cells. Method: CIK cells cultured from PBMCs of healthy subjects were stimulated with IL-21 and cellular viability and cytotoxicity to K562 cells were measured. To elucidate the mechanism of action of IL-21, mRNA expression of cytotoxic factors was assessed by RT-PCR and protein expression of significantly important cytotoxic factors and cytokine secretion were determined through flow cytometry and ELISA. Western blotting was performed to check the involvement of the JAK/STAT pathway following stimulation. Results: We found that IL-21 did not enhance in vitro proliferation of CIK cells, but did increase the number of cells expressing the CD3+/CD56+ phenotype. Cytotoxic potential was increased with corresponding increase in perforin ($0.9831{\pm}0.1265$ to $0.7592{\pm}0.1457$), granzyme B ($0.4084{\pm}0.1589$ to $0.7319{\pm}0.1639$) and FasL ($0.4015{\pm}0.2842$ to $0.7381{\pm}0.2568$). Interferon gamma and TNF-alpha were noted to increase ($25.8{\pm}6.1ng/L$ to $56.0{\pm}2.3ng/L$; and $5.64{\pm}0.61{\mu}g/L$ to $15.14{\pm}0.93{\mu}g/L$, respectively) while no significant differences were observed in the expression of granzyme A, TNF-alpha and NKG2D, and NKG2D. We further affirmed that IL-21 signals through the STAT-3 and STAT-5b signaling pathway in the CIK cell pool. Conclusion: IL-21 enhances cytotoxic potential of CIK cells through increasing expression of perforin, granzyme B, IFN-gamma and TNF-alpha. The effect is brought about by the activation of STAT-3 and STAT-5b proteins.

• Asian Pacific Journal of Cancer Prevention
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• v.17 no.4
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• pp.1999-2006
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• 2016

Background: Acute promyelocytic leukemia (APML) is characterized by the reciprocal translocation t(15;17) (p22;p12) resulting in the PML-$RAR{\alpha}$ fusion gene. A dual diagnostic and follow up approach was applied including cytogenetic demonstration of the t(15;17) translocation and detection dg PML-$RAR{\alpha}$ chimeric transcripts by molecular means. Purpose: Conventional cytogenetics involving bone marrow is beset with high probability of poor metaphase index and was substituted with phytohemagglutinin (PHA)-induced peripheral blood culture based cytogenetic analysis as a diagnostic & follow up modality in APML patients of Kashmir (North India). Both qualitative (RT-PCR) and quantitative (Q-PCR) tests were simultaneously carried out to authenticte the modified cytogenetics. Materials and Method: Patient samples were subjected to the said techniques to establish their baseline as well as follow-up status. Results: Initial cytogenetics revealed 30 patients (81%) Positive for t(15;17) whereas 7 (19%) had either cryptic translocation or were negative for t(15;17). Two cases had chromosome 16q deletion and no hallmark translocation t(15;17). Q-PCR status for PML-$RAR{\alpha}$ was found to be positive for all patients. All the APML patients were reassessed at the end of consolidation phase and during maintenance phase of chemotherapy where 6 patients had molecular relapse, wherein 4 also demonstrated cytogenetic relapse. Conclusions: It was found that PHA-induced peripheral blood cytogenetics along with molecular analysis could prove a reliable modality in the diagnosis and assessment of follow up response of APML patients.

• Research in Plant Disease
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• v.19 no.1
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• pp.12-20
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• 2013

To isolate antagonistic bacteria against sclerotinia rot of lettuce, caused by Sclerotinia sclerotiorum, soil samples were collected from the diseased greenhouse field in Namyangju city, Gyeong-gi province from 2007 to 2008. A total of 196 bacterial isolates were isolated using serial dilution method. In dual culture assay in vitro, 26 isolates showed more than 80% of inhibition rates of mycelial growth of S. sclerotiorum. Based on 16S rDNA sequence analysis, the 26 isolates were identified as Bacillus megaterium, B. cereus, B. subtilis, Arthrobacter nicotianae, A. ramosus, Pseudomonas filiscindens, Stenotrophomonas maltophilia, Brevibacterium frigoritolerans and Sphingobacterium faecium. The 26 isolates inhibited the mycelial growth of S. sclerotiorum up to 80% and the sclerotial germination 0-100%. In the greenhouse pot test of ten isolates conducted in summer, 2 isolates B. megaterium (DK6) and B. cereus (C210) showed control efficacy on sclerotia viability of S. sclerotiorum, 20% and 35%, respectively. In the greenhouse pot test in winter, the disease incidence of the control group was 80%, whereas those of 9 isolates among 26 were approximately 20%. From the result, the 9 isolates are expected as potentially antagonistic bacteria for biological control of sclerotinia rot of lettuce caused by S. sclerotiorum.

• Journal of Life Science
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• v.15 no.3 s.70
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• pp.337-343
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• 2005

Tapioca alcoholic distillery waste was utilized as dual purposes to produce citric acid and to reduce the amount of waste to be treated. Primarily an attempt was made to optimize the process conditions by Aspergillus niger in shake bath. The effects of pH, temperature, nitrogen and phosphorus sources on citric acid production were investigated. Maximum concentration of citric acid was made at temperature of $30^{\circ}C$ and pH of 4.3, while maximum cell dry weight was obtained at $35^{\circ}C$. The addition of methanol or ethanol to culture medium promoted citric acid production remarkably, but the addition of $NH_4NO_3,\;KH_2PO_4$ and Manganese as mineral source decreased the acid production.

• The Korean Journal of Mycology
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• v.43 no.4
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• pp.253-259
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• 2015

Anthracnose caused by Collectotrichum acutatum is the most devastating disease of pepper plants in Korea. In this study, we evaluated the effect of selected antagonistic bacteria on control of anthracnose and plant growth promotion of pepper plants under field conditions. Four different bacterial isolates used in the current study were isolated from the pepper rhizosphere (GJ01, GJ11) and tidal flat (LB01, LB14) in previous studies. Four bacterial isolates, together with a control strain (EXTN-1), showed antifungal activity against C. acutatum in a dual culture assay. To test for plant growth promotion effect, seedling vigor index and growth parameters of pepper were measured under field condition. As a result, all four bacterial isolates were effective for improving plant growth promotion. The strain GJ01 was the most effective in improving the seedling vigor on pepper, but the strain GJ11 in increasing the pepper fruit yield. The incidence of anthracnose was inhibited in the range of 63.2~72.5% by treatment of four bacterial isolates. The current study indicated that the four bacterial isolates could be used as potential biological control agents of anthracnose disease of pepper.

• Journal of Ginseng Research
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• v.40 no.4
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• pp.315-324
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• 2016

Background: Biocontrol agents are regarded as promising and environmental friendly approaches as agrochemicals for phytodiseases that cause serious environmental and health problems. Trichoderma species have been widely used in suppression of soil-borne pathogens. In this study, an endophytic fungus, Trichoderma gamsii YIM PH30019, from healthy Panax notoginseng root was investigated for its biocontrol potential. Methods: In vitro detached healthy roots, and pot and field experiments were used to investigate the pathogenicity and biocontrol efficacy of T. gamsii YIM PH30019 to the host plant. The antagonistic mechanisms against test phytopathogens were analyzed using dual culture, scanning electron microscopy, and volatile organic compounds (VOCs). Tolerance to chemical fertilizers was also tested in a series of concentrations. Results: The results indicated that T. gamsii YIM PH30019 was nonpathogenic to the host, presented appreciable biocontrol efficacy, and could tolerate chemical fertilizer concentrations of up to 20%. T. gamsii YIM PH30019 displayed antagonistic activities against the pathogenic fungi of P. notoginseng via production of VOCs. On the basis of gas chromatography-mass spectrometry, VOCs were identified as dimethyl disulfide, dibenzofuran, methanethiol, ketones, etc., which are effective ingredients for antagonistic activity. T. gamsii YIM PH30019 was able to improve the seedlings' emergence and protect P. notoginseng plants from soil-borne disease in the continuous cropping field tests. Conclusion: The results suggest that the endophytic fungus T. gamsii YIM PH30019 may have a good potential as a biological control agent against notoginseng phytodiseases and can provide a clue to further illuminate the interactions between Trichoderma and phytopathogens.

• Journal of Microbiology and Biotechnology
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• v.26 no.11
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• pp.1881-1890
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• 2016

Manganese superoxide dismutase (MnSOD) is a vital enzyme that protects cells from free radicals through eliminating superoxide radicals ($O^{2-}$). Hirudin, a kind of small active peptide molecule, is one of the strongest anticoagulants that can effectively cure thrombus diseases. In this study, we fused Hirudin to the C terminus of human MnSOD with the GGGGS linker to generate a novel dual-feature fusion protein, denoted as hMnSOD-Hirudin. The hMnSOD-Hirudin gene fragment was cloned into the pET15b (SmaI, CIAP) vector, forming a recombinant pET15b-hMnSOD-Hirudin plasmid, and then was transferred into Escherichia coli strain Rosetta-gami for expression. SDS-PAGE was used to detect the fusion protein, which was expected to be about 30 kDa upon IPTG induction. Furthermore, the hMnSOD-Hirudin protein was heavily detected as a soluble form in the supernatant. The purification rate observed after Ni NTA affinity chromatography was above 95%. The hMnSOD-Hirudin protein yield reached 67.25 mg per liter of bacterial culture. The identity of the purified protein was confirmed by western blotting. The hMnSOD-Hirudin protein activity assay evinced that the antioxidation activity of the hMnSOD-Hirudin protein obtained was $2,444.0{\pm}96.0U/mg$, and the anticoagulant activity of the hMnSOD-Hirudin protein was $599.0{\pm}35.0ATU/mg$. In addition, in vitro bioactivity assay showed that the hMnSOD-Hirudin protein had no or little cytotoxicity in H9c2, HK-2, and H9 (human $CD_4{^+}$, T cell) cell lines. Transwell migration assay and invasion assay showed that the hMnSOD-Hirudin protein could suppress human lung cancer 95-D cell metastasis and invasion in vitro.