• The Korean Journal of Nuclear Medicine
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• v.25 no.2
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• pp.252-258
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• 1991

With HMPAO we have synthesized in our laboratory, we labelled $^{99m}Tc$ to canine leukocytes. Experimental abscess made by subcutaneous injection with Staphylococcus aureus was imaged with these $^{99m}Tc$ labelled leukocytes. Labelling efficiency of HMPAO with $^{99m}Tc$ was $66.2%{\pm}14.6%$ (N=9). Labelling efficiency of leukocytes with $^{99m}Tc-HMPAO$ was $54%{\pm}7.7%$ (N=7). Cell bound radioactivity in $^{99m}Tc-HMPAO$ labelled leukocytes was around 80% when these cells were incubated in plasma in vitro at $37^{\circ}C$ for 5 hours. In vivo cell bound activity was over 80% at 24 hours after injection. One day and four days after inoculation, uptake at the inflammatory focus was found with $^{99m}Tc$ labelled leukocytes. Uptake showed up in 4 hour image, and the uptake at the lesion was most prominent in 24 hour image. These findings show that in-house-synthesized HMPAO could be used for labelling leukocytes with $^{99m}Tc$, and that s$^{99m}Tc-HMPAO-labelled$ leukocytes were so stable and viable that inflammatory focus could be visualized with these $^{99m}Tc-labelled$ leukocytes.

• Journal of Microbiology and Biotechnology
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• v.13 no.6
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• pp.881-891
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• 2003

Paenibacillus polymyxa is a plant growth-promoting rhizobacterium (PGPR) which can be used for biological control of plant diseases. Several bacterial strains were isolated from rotten roots of Korean ginseng (Panax ginseng C. A. Meyer) that were in storage. These strains were identified as P. polymyxa, based on a RAPD analysis using a P. polymyxa-specific primer, cultural and physiological characteristics, an analysis utilizing the Biolog system, gas chromatography of fatty acid methyl esters (GC-FAME), and the 16S rDNA sequence analysis. These strains were found to cause the rot in stored ginseng roots. Twenty-six P. polymyxa strains, including twenty GBR strains, were phylogenetically classified into two groups according to the ERIC and BOX-PCR analyses and 16S rDNA sequencing, and the resulting groupings systematized to the degrees of virulence of each strain in causing root rot. In particular, highly virulent GBR strains clustered together, and this group may be considered as subspecies or biovar. The virulence of the strains seemed to be related to their starch hydrolysis enzyme activity, but not their cellulase or hemicellulase activity, since strains with reduced or no starch-hydrolytic activity showed little or no virulence. Artificial inoculation of the highly virulent strain GBR-1 onto the root surfaces of Korean ginseng resulted in small brown lesions which were sunken and confined to the outer portion of the root. Ginseng root discs inoculated in vitro or two-year-old roots grown in soil drenched with the inoculum developed significant rot only when the inoculum density was $10^{6}-10^{7}$ or more colony-forming units (CFU) per ml. These results suggest that P. polymyxa might induce ginseng root rot if their population levels are high. Based on these results, it is recommended that the concentration of P. polymyxa should be monitored, when it is used as a biocontrol agent of ginseng, especially in the treatment of stored roots.

• Journal of Microbiology and Biotechnology
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• v.28 no.11
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• pp.1834-1845
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• 2018

The lactobacilli associated with a fermented goat milk product from Tajikistan were isolated to characterize their technological properties and antibiotic resistances in order to assess their suitability for development as starter cultures. In this study, twenty three strains were identified by 16S rRNA sequencing as typical dairy-associated lactic acid bacterial strains, i.e. L. plantarum, L. pentosus, L. delbrueckii, L. helveticus and L. paracasei. These strains were generally susceptible to most antibiotics tested in this study and this allowed a selection of strains as safe starters. The draft genomes of four representative strains were sequenced and the number of contigs of the four assembled genomes ranged from 51 to 245 and the genome sizes ranged from 1.75 to 3.24 Mbp. These representative strains showed differences in their growth behavior and pH-reducing abilities in in vitro studies. The co-inoculation of these Lactobacillus spp. strains together with a yeast Kluyveromyces marxianus MBT-5698, or together with the yeast and an additional Streptococcus thermophilus MBT-2, led to a pH reduction to 3.4 after 48 h. Only in the case of fermentation inoculated with the co-culture, the viscosity of the milk increased noticeably. In contrast, fermentations with single strains did not lead to gelation of the milk or to a decrease in the pH after 24h. The results of this study provide a comprehensive understanding of the predominant lactobacilli related to Tajikistani fermented milk products.

• Journal of Plant Biotechnology
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• v.1 no.1
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• pp.1-7
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• 1999

Large-scale cultures of plant cell, tissue, and organ have been achieved by using BTBB. When different sized BTBBs (5 L, 20 L, 100 L, 300 L, and 500 L) were tested for the culture of yew cells (Taxus cuspidata Sieb. et Zucc.), cell growth increment reached to 94.5% in SCV after 24 days of culture with 30% of inoculation cell density. However, there were some variations in the production of taxol and its derivatives among the BTBBs of different size. Approximate 4 ㎎/l of taxol and 84 ㎎/l of total taxanes were obtained by using a 500L BTBB after 6 weeks of culture. With a 20L BTBB, about 20,000 cuttings of virus-free potatoes (cv. Dejima) could be obtained by inoculating 128 explants and maintaining 8 weeks under 16 hr light illumination. The frequency of ex vitro rooting of the cuttings revealed as more than 99% under 30% shade. By incorporating two-stage culture process consisting of multiple bulblet formation in solid medium and bulblet development in liquid medium, mass propagation of lily through bioreactor seemed to be possible. In the case of 'Marcopolo', the growth of mini-bulblets in BTBB was nearly 10 folds faster than that of the solid medium. Time course study revealed that maximum MAR yield of ginseng (Panax ginseng C. A. Meyer) in a 5 L and 20 L BTBB after 8 weeks of culture was 500 g and 2.2 ㎏, respectively. By cutting the MAR once and/or twice during the culture, the yield of root biomass could be increased more than 50% in fresh weight at the time of harvest. With initial inoculum of 500 g of sliced MAR in a 500 L BTBB, 74.8 ㎏ of adventitious root mass was obtained after 8 weeks of culture. The average content of total ginseng saponin obtained from small-scale and/or pilotscale BTBBs was approximately 1% per gram dry weight. Based on our results, we suggest that large-scale cultures of plant cell, tissue, and organ using BTBB system should be quite a feasible approach when compared with conventional method of tissue culture.

• The Plant Pathology Journal
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• v.29 no.4
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• pp.386-396
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• 2013

Reactive oxygen species (ROS) generation in tomato plants by Ralstonia solanacearum infection and the role of hydrogen peroxide ($H_2O_2$) and nitric oxide in tomato bacterial wilt control were demonstrated. During disease development of tomato bacterial wilt, accumulation of superoxide anion ($O_2{^-}$) and $H_2O_2$ was observed and lipid peroxidation also occurred in the tomato leaf tissues. High doses of $H_2O_2$ and sodium nitroprusside (SNP) nitric oxide donor showed phytotoxicity to detached tomato leaves 1 day after petiole feeding showing reduced fresh weight. Both $H_2O_2$ and SNP have in vitro antibacterial activities against R. solanacearum in a dose-dependent manner, as well as plant protection in detached tomato leaves against bacterial wilt by $10^6$ and $10^7$ cfu/ml of R. solanacearum. $H_2O_2$- and SNP-mediated protection was also evaluated in pots using soil-drench treatment with the bacterial inoculation, and relative 'area under the disease progressive curve (AUDPC)' was calculated to compare disease protection by $H_2O_2$ and/or SNP with untreated control. Neither $H_2O_2$ nor SNP protect the tomato seedlings from the bacterial wilt, but $H_2O_2$ + SNP mixture significantly decreased disease severity with reduced relative AUDPC. These results suggest that $H_2O_2$ and SNP could be used together to control bacterial wilt in tomato plants as bactericidal agents.

• Journal of Mushroom
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• v.9 no.3
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• pp.110-115
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• 2011

Trichoderma disease of oyster mushroom has not been effectively detected in the field for testing its resistance against the disease with its varieties. In this study, we investigated the methods to detect its resistance in the laboratory by using media, which enables us to understand the relevant characteristics (e.g., lysis, toxin enzyme, mycelial growth rate). In coculturing with strains of Trichoderma and oyster mushroom, it is possible to observe the difference in the resistance of oyster mushroom against Trichoderma with the phenomena of barrage reaction, overgrowth and lysis. We also observed the inhibition of mycelial growth of oyster mushroom using the dilution method with 48-well plate, but could not observed the inhibition of mycelial growth using the filter paper method of cultural supernatant. In simultaneously culturing both Trichoderma and oyster mushroom, it was possible to detect the inhibition of the mycelial growth of oyster mushroom, but Trichoderma mycelium did not overgrow against oyster mushroom. We found that the pathogenicity was efficient in using solid medium with the phenomena of overgrowth and lysis by inoculating Trichoderma on top of mycelia of oyster mushroom. In conclusion, the methods (e.g., coculture method, dilution method with 48-well plate, post-inoculation method) are recommended to detect the resistance of oyster mushroom against Trichoderma disease.

• Parasites, Hosts and Diseases
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• v.33 no.4
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• pp.349-356
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• 1995

We have investigated the action of sodium nitrite on the growth and morphologic changes of T uaginolis and on the treatment of subcutaneous abscess by trichomonad in mice. Sodium nitrite inhibited the growth of metronidazole-sensitive KT9 isolate and metronidazole-resistant CDC85 strain of T vcsinalis as concentration of 6 mM and 10 mM respectively Intraperitoneal injection of sodium nitrite (70 ㎍, 100 ㎍, 130 ㎍/g body weight) did not reduce the size of abscess produced by subcutaneous inoculation of T uasinnlis in mice. T uosinnlis, treated with sodium nitrite at concentration giving about 50% inhibition of growth, showed fissures, many vacuoles and electron-translucent zone in the cytoplasm by transmission electron microscopy. In the case of CDC85 treated with 9 mM sodium nitrite, hydrogenosomal matrical change , destruction of hydrogenosomal membrane, autophagic vacuoles, disappearance of Golgi complex and polysome were notably observed . With above results, it is assumed that sodium nitrite inhibits the growth of metronidazole-sensitive and - resistant strains of T. ucsinalis and induces the morphological changes of 7 uusinalis although it does not affect in reducing of abscess size by vagiginalis in mice.

• Microbiology and Biotechnology Letters
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• v.35 no.1
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• pp.1-10
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• 2007

The 44 endophytic bacterial strains were isolated from surface-sterilized root of rice cultivated in seven different locations of Chungcheong province, Korea. Each isolate was introduced into rice seedlings grown gnotobiotically by inoculating scissor-cut first true leaf with cell suspensions, and the colonization capacity of each isolate in root tissue was analyzed at 7 days after inoculation. Sixteen out of 44 isolates were re-isolated from root successfully with the frequency of $10^{3-5}$ CFU/g tissue. Interestingly, seven out of 16 isolates were identified as Burkholderia species. The identity between inoculated strains and re-isolates was confirmed by genomic finger-printing and 16S rDNA sequence analysis. By a confocal laser scanning microscopic observation it was revealed that KJ001 strain, one of the sixteen isolates tagged with gfp colonized in root tissue especially around xylem. Six out of seven Burkholderia strains obtained in this study showed antagonizing activities against seven different fungal pathogens, contain nifH gene, and five of them enhanced growth of cucumber over 30%. The isolates showed no hypersensitive response on tobacco leaves and no pathogenecity in rice. From these results it was found that the endophytic Burkholderia strains will be useful in agriculture to develop a biocontrol agent or a bio-fertilizer.

• Asian-Australasian Journal of Animal Sciences
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• v.30 no.7
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• pp.950-956
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• 2017

Objective: Effects of newly isolated Lactobacillus plantarum on the fermentation and chemical composition of fresh rice straw silage was evaluated in this study. Methods: Lactic acid bacteria (LAB) from good crop silage were screened by growing them in MRS broth and a minimal medium with low carbohydrate content. Selected LAB (LAB 1821) were Gram-positive, rods, catalase negative, and were identified to be Lactobacillus plantarum based on their biochemical characteristics and a 16S rRNA analysis. Fresh rice straw was ensiled with two isolated LAB (1821 and 1841), two commercial inoculants (HM/F and P1132) and no additive as a control. Results: After 2 months of storage at ambient temperature, rice straw silages treated with additives were well-preserved, the pH values and butyric and acetic acid contents were lower, and the lactic acid content and lactic/acetic acid ratio were higher than those in the control (p<0.05). Acidity (pH) was lowest, and lactic acid highest, in 1821-treated silage (p<0.05). The $NH_3-N$ content decreased significantly in inoculant-treated silage (p<0.05) and the $NH_3-N$ content in 1821-treated silage was lowest among the treatments. The dry matter (DM) content of the control silage was lower than that of fresh rice straw (p<0.05), while that of the 1841- and p1174-inoculant-treated silages was significantly higher than that of HM/F-treated silage. Microbial additives did not have any significant (p>0.05) effect on acid detergent fiber or neutral detergent fiber contents. Crude protein (CP) content and in vitro DM digestibility (IVDMD) increased after inoculation of LAB 1821 (p<0.05). Conclusion: LAB 1821 increased the CP, IVDMD, lactic acid content and ratio of lactic acid to acetic acid in rice straw silage and decreased the pH, acetic acid, $NH_3-N$, and butyric acid contents. Therefore, adding LAB 1821 improved the fermentation quality and feed value of rice straw silage.

• The Journal of Korean Obstetrics and Gynecology
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• v.27 no.4
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• pp.1-14
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• 2014

Objectives: This study was designed to investigate the anti-tumor metastasis by anti-inflammatory and innate immunomodulating effects of extracts of Jipae-san on cancer cells. Methods: Antimetastatic experiments were conducted in vivo mouse model by using 4T1 mouse mammary carcinoma cells. Cell viability of Jipae-san was tested with 4T1 mouse mammary carcinoma cells, colon 26-M3.1 carcinoma cells and macrophage. In addition expression of $TNF-{\alpha}$ and NO induced by LPS was measured after treating with Jipae-san. To observe innate immunomodulating effects of Jipae-san on macrophage, we measured $TNF-{\alpha}$, IL-12, IL-6 and MCP-1, respectively. Cell cytotoxicity was tested with the macrophage stimulated with Jipae-san and we evaluated the activation of $TNF-{\alpha}$ and NO. And the effect of Jipae-san on metastasis was measured without NK-cell using GM1 serum. Results: Intravenous inoculation of Jipae-san significantly inhibited metastasis of 4T1 mouse mammary carcinoma cells. In an in vitro cytotoxicity analysis, cell growth are closer to 100% less than $1,000{\mu}g/ml$ concentration. The expression of $TNF-{\alpha}$ and NO induced by LPS after treating Jipae-san was down regulated in dose-dependent manner. Level of cytokines such as $TNF-{\alpha}$, IL-12, IL-6 and MCP-1 of Jipae-san group were up regulated in compared to the control group. The macrophage stimulated with Jipae-san significantly inhibits the cancer cell at ratio of 10:1, 20:1. The activation of NO was significantly up regualted in a group of 5:1, 10:1, 20:1. The depletion of NK-cells by anti-asialo GM1 serum partly abolished the inhibitory effect of Jipae-san on tumor metastasis. Conclusions: Jipae-san appears to have considerable activity on the anti-metastasis by inflammation control and activation of innate immune system.