• Journal of the Korea Institute of Military Science and Technology
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• v.16 no.3
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• pp.390-397
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• 2013

Primary-cultured immune cells are widely used in research to elucidate the mechanism of inflammation including chemotaxis, production of reactive oxygen species, cytokine release and antigen presenting. Mice are one of the species of experimental animals commonly used for such studies. Immune cells can be isolated and cultured from various organs such as bone marrow, peritoneal cavity, lung, spleen. For elaborated experimental studies, immune cells should be elicited with inflammatory substances or proliferated in vitro with special media. This paper details methods of obtaining immune cells from various organs of mice and investigating immune mechanism using isolated immune cells. It contains standard protocols of isolating and culturing immune cells from bone marrow, peritoneal cavity and lymphoid organs. It also covers the methods of investigating immune mechanism such as ELISA, western blotting, confocal microscopy and ELISPOT assay. With the works in this study, we established the standardized isolation and analysis methods of primary-cultured immune cells.

• Journal of Microbiology and Biotechnology
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• v.17 no.1
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• pp.168-175
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• 2007

We report here on the cultivation of numerous novel bacterial species from a eutrophic freshwater pond. A total of 60 strains, 15 strains per each culture medium, were obtained from the surface of a eutrophic freshwater pond by employing a conventional dilution-plating method with four different kinds of culture media, including R2A, 1/10R2A, PCA, and 1/10PCA. Among the 60 strains isolated, 27 strains showed less than 97% 16S rRNA gene sequence similarities to validly published species, and thus they are considered to comprise 19 novel species. Of the 27 strains assigned to the novel species, the majority of the strains (20 strains) were affiliated with the Alphaproteobacteria and Betaproteobacteria. The remaining 7 strains were affiliated with the Gammaproteobacteria, Firmicutes, Actinobacteria, and Deinococci. Because we have isolated 19 novel species from a usual freshwater pond using a conventional culturing technique, our results suggest that an unexplored ecosystem, even if it looks like a common ecosystem found elsewhere, harbors diverse unidentified microbes, which will be definitely further characterized.

• Korean Journal of Microbiology
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• v.13 no.3
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• pp.101-108
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• 1975

Physiological effects of 2,4-D on the growth of Chlorella ellipsoidea were investigated culturing the alage in the MN4 media containing 0. $10^{-4}/M$ and $4<\times}10^{-4}M$ 2,4-D. During 6 days culture were taken to analysis with respect to overall growth, photosynthesis, respiration and chemical composition. Results obtained from the experiment were as follows : 1) The growth of chlorella was increased at $10^{-4}M$ and decreased at $4{\times}10^{-4}M$ of 2,4-D concentrations 2) At $10^{-4}M$ pf 2,4-D cpncentration, the activity of photosynthesis enhanced relative to contro. while at $4{\times}10^{-4}M$ it was not changed. In both concentrations, however, the rate of respiration was down from the control. 3) At $10^{-4}M$ 2,4-D, the concentration of carbondrate metabolites was not changed relative to control, while significant increase in the concentrations of proteins and nucleic acids was observed. On the other hand at $4{\times}10^{-4}M$ of 2,4-D concentrations, all the metabolites including carbohydrates, proteins and nucleic acids were descreased. 4) It is concluded that 2,4-D at $10^{-4}M$ concentration accelerates the growth of chlorella by promoting the activities of photosynthesis and biosynthesis of proteins and nucleic acids.

• Journal of Korean Academy of Fundamentals of Nursing
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• v.16 no.2
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• pp.207-213
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• 2009

Purpose: This study was done to examine how medication contamination in a single-dose glass ampule is affected by minute glass flakes generated in different methods of cutting the ampule. Method: Sixty medication-containing glass ampules were randomly assigned to two groups. The number of glass flakes, resulting from two different cutting methods (with cotton and without cotton), were counted under the microscope. Contamination was evaluated by extracted the medication with a syringe and culturing it in E. coli, coliform, and aerobic bacteria culture media. Result: Fewer glass flakes were found in the ampules when the ampule was cut with cotton. The use of cotton, however, did not significantly change the degree of drug contamination. Conclusion: Although minute glass flakes generated in the ampule cutting operation did not significantly contaminate the medication and the use of cotton decreased the number of glass flakes in the ampules, glass flakes injected into the blood and tissues of the patient remain a risk factor. Therefore, pre-filled syringes or syringes with filters would be alternative methods and safeguards against the possible injection of glass flakes generated while cutting the ampule.

• Journal of the Korean Society of Tobacco Science
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• v.14 no.1
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• pp.3-11
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• 1992

The present study examined various environmental and cultural media conditions for in vitro "rescue" of cross-fertilized ovules formed through sexual crosses between Nicotiana rustica and N. tabacum cv. BY4. The response ovules to two cultural procedures was compared; ovules were cultured either separately or left attached to the placenta. Total yield of seedlings and percent of normal seedlings were increased by culturing individual ovules separately, rather than on excised placenta. Total yield of seedlings and number of normal seedlings were produced following in vitro culture of individual fertilized ovules of N. rustica X M tabacum cv. BY4 at four days post-pollination on NN medium containing 2% sucrose. In the in vitro culture of fertilized ovules, high sucrose concentration increased the frequency of seedlings of abnormal appearance. Therefore, sucrose should be supplied to developing ovules at gradually decreased concentrations. Culture of fertilized ovules from three to eight days after pollination gave increased number of seedlings, but with delayed cultral time the number of morphologically normal seedling were decreased. Hybrids were uniform in appearance and showed vegetative heterosis but flower characteristics were generally intermediate between those of the parents. All hvbrids evaluated were self-sterile.f-sterile.

• Journal of Food Hygiene and Safety
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• v.13 no.1
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• pp.20-23
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• 1998

The objective of this study was to evaluate the bifidogenic effect and a-glucosidase inhibitory effect of artificially cultured Phellinus linteus. The water extract of P. linteus promoted the growth of Bifidobacterium breve as well as the decrease of final pH in the media culturing intestinal bacteria. The growth of lactic acid bacteria inhibited effectively the bacterial enzymes, ${\beta}-glucosidase,\;{\beta}-glucuronidase$ and tryptophanase, of intestinal bactetria. The water extract of P. linteus inhibited maltase, sucrase and ${\alpha}-amylase$ of rat intestine.

• Korean Journal of Soil Science and Fertilizer
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• v.49 no.5
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• pp.461-468
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• 2016

One of the important phytohormones produced by plant growth promoting bacteria is the auxin; indole-3-acetic acid (IAA), with L-tryptophan as the precursor. In this study, we focused on the investigation of optimal conditions for the production of IAA by Bacillus megaterium BM5. We investigated culturing conditions, such as incubation temperature, pH of the culture medium and incubation period, with varying media components such as inoculation volume, tryptophan concentration and carbon and nitrogen source. Besides, optimization study intended for high IAA production was carried out with fermentation parameters such as rpm and aeration. The initial yield of $42{\mu}g\;IAA\;ml^{-1}$ after 24 hr increased to $85{\mu}g\;ml^{-1}$ when 5% (v/v) of L-tryptophan was used in the culture broth. The maximum yield of $320{\mu}g\;IAA\;ml^{-1}$ was observed in trypticase soy broth (TSB) supplemented with starch and soybean meal as C and N sources with a C/N ratio of 3:1 (v/v) at $30^{\circ}C$, pH 8.0 for 48 hrs with 1.0 vvm and 250 rpm in 5 L working volume using 10 L scale fermenter. The bacterial auxin extracted from the culture broth was confirmed by thin layer chromatography and high-performance liquid chromatography and effect on plant growth was confirmed by root elongation test.

• Journal of Microbiology
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• v.33 no.4
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• pp.328-333
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• 1995

$\^$31/ P-NMR experiment was performed to detect phophonates (Pn) utilization and degradation in the several different C-P lyase mutants of E. coli and in E. aerogenes and the recombinants. The relative peak intensity (RPI) for the standard samples of 0.5 mM methylphosphonate (MPn) and 1.0 mM aminoethylphosphonate in glucose-MOPS medium showed 0.5 : 1.0 ratio. In the case of BW14329 (.DELTA.phnC-P, .delta.phoA), RPI did not change significantly after 24 hrs culturing, which means it nearly could not utilize Pn. In vivo $\^$31/ P-NMR spectrum of E. aerogens (BWKL 16627) during 3 hrs starvation showed two intense peaks at 0-2 ppm and at near-10 ppm which indicate intracellular orthophosphate (Pi) and pyrophosphate (PPi), respectively. Both of them might be released by degradation of inorganic polyphosphate pool. When MPn is supplied to the medium as an unique P source, Pi content in the cell has the constant, but PPi seems to be slightly decreased. Recombinants (BWKL 16954) grew slower than E. aerogenes in the glucose-MOPS media with various P sources. In vivo $\^$31/ P-NMR spectrum of recombinant did not show any intense signal in the cell. Surprisingly, under the cultivation adding with MPn, a few intense peaks in the region of Pi AND phospate monoester were detected.

• Journal of Microbiology
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• v.45 no.3
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• pp.199-204
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• 2007

The objective of this study was to evaluate the effect of soluble carbohydrates (glucose, cellobiose), pH (6.0, 6.5, 7.0), and rumen microbial growth factors (VFA, vitamins) on biohydrogenation of linoleic acid (LA) by mixed rumen fungi. Addition of glucose or cellobiose to culture media slowed the rate of biohydrogenation; only 35-40% of LA was converted to conjugated linoleic acid (CLA) or vaccenic acid (VA) within 24 h of incubation, whereas in the control treatment, 100% of LA was converted within 24 h. Addition of VFA or vitamins did not affect biohydrogenation activity or CLA production. Culturing rumen fungi at pH 6.0 slowed biohydrogenation compared with pH 6.5 or 7.0. CLA production was reduced by pH 6.0 compared with control (pH 6.5), but was higher with pH 7.0. Biohydrogenation of LA to VA was complete within 72 h at pH 6.0, 24 h at pH 6.5, and 48 h at pH 7.0. It is concluded that optimum conditions for biohydrogenation of LA and for CLA production by rumen fungi were provided without addition of soluble carbohydrates, VFA or vitamins to the culture medium; optimum pH was 6.5 for biohydrogenation and 7.0 for CLA production.

• Journal of Microbiology
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• v.37 no.4
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• pp.200-205
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• 1999

7-Aminocephalosporanic acid (7-ACA) is the initial compound in preparation of cephalosporin antibiotics widely used in clinical treatment. Bacteria producing glutaryl 7-ACA acylase, which convert cephalosporin C to 7-ACA, has been screened in soil samples. A bacterial strain exhibiting high glutaryl 7-ACA acylase activity, designated KAC-1, was isolated and identified as a strain of Pseudomonas diminuta by characterizing its morphological and physiological properties. The screening procedures include culturing on enrichment media containing glutaric acid, glutamate, and glutaryl 7-aminocephalosporanic acid as selective carbon sources. To enhance enzyme production, optimal cultivation conditions were investigated. This strain grew optimally at pH 7 to 9 and in temperatures of 20 to 40 C, but acylase production was higher when the strain was grown at 25 C. Glutaric acid, glutamate and glucos also acted as inducers for acylase production. In a jar fermenter culture, P. diminuta KAC-1 produce acylase in a growth-associated manner. The substrate specificity of KAC-1 acylase by cell extract showed that this enzyme had specificity toward glutaryl 7-ACA, glutaryl 7-ADCA, but not cephalosporin C.