• Proceedings of the Korean Society of Applied Pharmacology
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• 1994.04a
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• pp.131-136
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• 1994

In the present study, in order to make an in vitro model of gastric mucosa for physiological and pharmacological studies, we established two immortalized gastric surface mucous cell lines (GSM06 and GSM10), which produce periodic acid-Schiff (PAS)-and concanavalin A (Con A)-positive glycoproteins, from a primary culture of gastric fundic mucosal cells of adult transgenic mice harboring a temperature-sensitive simian virus 40 large T-antigen gene 〔1]. Gastric fundic mucosal cells were isolated as a modification of a previously described method for rats by Schepp et al. (2). The isolated gastric fundic mucosal cells were cultured in DME/F12 medium supplemented with 2% fetal bovine serum (FBS), 1% ITES (consisting of 2 mg/1 insulin, 2 mgg/1 transferrin, 0.122 mg/1 ethanolamine and 0.00914 mg/1 sodium selenite) and 10 ng/ml recombinant epidermal growth factor (EGF) in a collagen-coated culture dish. To remove fibroblastic cells from the culture, gastric mucosal cells were incubated in the culture medium containing dispase (25 U/ml) for 24 h. The cells, uncontaminated with fibroblastic cells, were then cloned by colony formation. In our series of three attempts, two cell lines (GSM06 and GSM10) have been established at last. The cells proliferated, attached to the dish ana grew until confluent monolayers were formed, and maintained tight contact with neighboring cells. Both GSM06 and GSM10 cells have now been in culture for more than 9 months with regular passaging. The either cell produced

• Korean Journal of Clinical Laboratory Science
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• v.42 no.2
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• pp.86-91
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• 2010

We evaluated the performance of a novel screening test, PBP2a MRSA rapid kit (Dinona Inc., Iksan, Korea), for methicillin-resistant Staphylococcus aureus (MRSA) based on a immunochromatographic assay. The test is able to detect penicillin-binding protein 2a (PBP2a) using the nasal specimens from health care workers. The nasal specimens were obtained from 69 healthcare workers and were incubated in enrichment broth followed eight hours incubatin in BHI with cefoxitin $4{\mu}g/mL$. These broth were tested by PBP2a Rapid Kit. The enrichment broths were also directly tested for tube coagulase using the conventional identification method. 19 of 22 MRSA showed positive results by PBP2a rapid test and direct coagulase test (the sensitivity for detection of MRSA, 86.36%). While, 8 of 47 non-MRSA showed false positive results for the two tests. All of the 8 non-MRSA which showed false positive were co-colonizing isolates with MRCNS and MSSA. In addition, 46 of 49 methicillin-resistant staphylococci (MRS) showed positive results for PBP2a MRSA rapid kit (the sensitivity for detection of MRS, 93.8%), and all of 20 non-MRS showed negative results (specificity, 100%). The combination of PBP2a MRSA rapid kit and direct coagulase test showed the good sensitivity for detection of MRSA from anterior nares but frequently showed false positive results from the co-colonizing carrier with MRCNS and MSSA.

• Journal of Pharmaceutical Investigation
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• v.31 no.3
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• pp.191-196
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• 2001

Biodegradable polymeric microspheres were studied for their usefulness as carriers for the delivery of vaccine antigens. However, protein antigen could be denatured during microencapsulation processes due to the exposure to the organic phase and stress condition of cavitation and shear force. Therefore this study was carried out to re-evaluate the degree of protein denaturation during microencapsulation with poly(lactide-co-glycolide) (PLGA) copolymer. PLGA microspheres containing ovalbumin (OVA), prepared by W/O/W multiple emulsification method, were suspended in pH 7.4 PBS and incubated with shaking at $37.5^{\circ}C$. Drug released medium was collected periodically and analyzed for protein contents by micro-BCA protein assay. In order to evaluate the protein integrity, release medium was subjected to the analyses of SDS-PAGE and size exclusion chromatography (SEC). And enzyme-linked immunosorbent assay (ELISA) was introduced to measure the immunoreactivity of entrapped OVA and to get an insight into the three-dimensional structure of epitope. The structures of entrapped protein were not affected significantly by the results of SDS-PAGE and SEC. However, immunoreactivity of released antigen was varied, revealing the possibility of protein denaturation in some microspheres when it was evaluate by ELISA method. Therefore, in order to express the degree of protein denaturation, antigenicity ratio (AR) was obtained as follows: amount of immunoreactivity of OVA/total amount of OVA released ${\times}100(%)$. ELISA method was an efficient tool to detect a protein denaturation during microencapsulation and the comparison of AR values resulted in more accurate evaluation for immunoreactivity of entrapped protein.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.42 no.6
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• pp.537-544
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• 2009

This study was conducted to improve yield and functional properties of autoclave-treated salmon frame extracts (SFETA) using commercial enzymes (Alcalase 2.4 L FG, Flavourzyme 500 MG, Neutrase 0.8 L and Protamex 1.5 MG). Yield and angiotensin I converting enzyme (ACE) inhibitory activity of all enzymatic hydrolysates improved compared to those of control (undigested extracts), which were the highest in hydrolysates incubated with Protamex 1.5 MG for 4 hrs (P4-treated hydrolysates) and 2 hrs (P2-treated hydrolysates), respectively. However, antioxidant activities of all enzymatic hydrolysates showed less than 29%. According to the trichloroacetic acid soluble-N, volatile component intensity and sensory evaluation, when compared to control, taste of P4-treated hydrolysates improved, while its fish odor strongly smelt. Therefore, for efficient use of P4-hydrolysates, the fish odor should be improved by Maillard reaction of extracts or pre-treatment of salmon frame.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.6
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• pp.842-849
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• 2007

The purpose of the present study was to produce live offspring from in vitro fertilized goat embryos. Oocytes were collected from abattoir ovaries and kept in oocyte collection medium. Oocytes were washed 4-5 times with maturation medium containing medium-199 with 5 ${\mu}g/ml$ FSH, 100 ${\mu}g/ml$ LH, 1 ${\mu}g/ml$ estradiol-$17{\beta}$ 50 ${\mu}g/ml$ gentamycin, 10% inactivated estrus goat serum, and 3% BSA (fatty acid free). Oocytes were placed in 100 ${\mu}l$ drops of maturation medium containing granulosa cell monolayer and incubated in a 5% $CO_2$ incubator at $38.5^{\circ}C$ for 27 h. For capacitation of spermatozoa fresh semen was processed and mixed in 3 ml fertilization TALP medium containing 50 ${\mu}g/ml$ heparin and kept in the above incubator for 2 h. The capacitated spermatozoa were coincubated with matured oocytes for fertilization. Cleaved embryos were separated and cultured in embryo development medium with oviductal cells and 494 embryos were produced. Recipient goats were synchronized with two injections of 15 mg $PGF_{{2}{\alpha}}$/goat 10 days apart. Eighty early stage embryos were transferred into the uterotubal junction of 14 surrogate mothers using laparoscopy techniques. One recipient delivered twin kids, whereas another two recipients each.delivered a single kid The parentage of these kids was evaluated using highly polymorphic co-dominant microsatellites markers. From the present study, it was concluded that live goat kids can be produced from in vitro matured and fertilized goat embryos, to the best of our knowledge for the first time in India.

• Microbiology and Biotechnology Letters
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• v.31 no.2
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• pp.191-196
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• 2003

This study was carried out to investigate the isolation, identification, and antibacterial activity of lactic acid bacteria related to kimchi fermentation. Diluted kimchi soup was plated on the MRS agar media with CaCO$_3$ and incubated at $25^{\circ}C$ for 2 days. A total of 27 strains of lactic acid bacteria from various indigenous, spontaneously fermented vegetables (kimchi) were isolated. Combined methods of Bergey's manual of systematic bacteriology, BPB media analysis and 16S rDNA sequence analysis were applied for identification, however, their results did not coincide in several cases. Isolated lactic acid bacteria could be classified by the 16S rDNA sequence analysis as Leuconostoc mesenteriodes, Leu. carnosum, Lactobacillus curvatus, Lac. pentosus, Weisselia kimchi, W. cibaria, and Pediococcus pentosaceus. Leu. carnosum has not been reported in kimchi lactic acid bacteria. In addition, antibacterial activities of the isolates were tested with Bacillus subtilis, Escherichia coli, Salmonella enteritidis, S. paratyphica, S. typhi, Staphylococcus aureus, Shigella boydii, and S. sonnei. Some of isolates showed significant antibacterial activities to those pathogens.

• Microbiology and Biotechnology Letters
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• v.20 no.4
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• pp.390-394
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• 1992

Some enzymatic characteristics of the aflatoxin degrading factor produced extraceIlularly by Aspergillus awamori var. fumeus were investigated. When aflatoxin B1 was incubated with the culture filtrate of A. awamori var. fumeus. 60% of it was degraded within an hour. The degradation rate decreased with time and there was virtually no degradation after one hour. The apparent Michaelis constant ($K_m$) determined by Lineweaver-Burk plot was $10.2{\mu}M$. The optimum degradation was observed at $30^{\circ}C$ and pH 5. For the degradation, molecular oxygen seemed to be required. The degradation was enhanced by the $Co^{2+}$. but was inhibited by many other ions like $Fe^{2+}$, $Ca^{2+}$. $Mg^{2+}$, $Zn^{2+}$,$Cu^{2+}$, and $Ba^{2+}$, The presence of either KeN or metyrapone inhibited the reaction while that of $NaI0_4$ cytochrome C or NADPH showed no effect.

• The Journal of the Korean dental association
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• v.46 no.9
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• pp.563-573
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• 2008

Purpose : The purpose of this study is to investigate the effects of Simvastain, which is HMG-CoA reductase inhibitor, on proliferation and differentiation of osteoblast. Materials & Methods : Twenty-four cell culture plates containing essential medium were seeded with UMR-106 cell lines, at density of 5 x $10^4$ cells per plate. Each plates were incubated with 5% $CO^2$incubator $37^{\circ}C$. Starting from 2 days after incubation, cell culture medias were replaced with Osteogenesis induction media every 2 days, for 12 days. In some plates, 0.01, 0.1, 1, 10, $100\muM$ of Simvastatin were added with Osteogenesis induction media, and classified as "test group". Those not added with Simvastatin were classified as "control group". Results : 1. When Alrizarin Red staining was observed with naked eye, control group showed normal deep red color, but test group show rapid decrease of red color as Simvastatin concentration increased more than $0.1\muM$. 2, When observed with microscope, compared to control group, amount of osteo matrix stained with Alrizarin Red decreased rapidly in Simvastatin concentration more than $0.1\muM$. 3. In optical density analysis, regarding control group as a basis, mineral deposition decreased rapidly when Simvastatin concentration increased more than $0.1\muM$. 4. In flow cytometry analysis, survival rate of UMR-106 cell showed no changes in both control group and test group. Conclusion : From the above results, we were able to identify that Simvastatin inhibited osteogenesis without effecting survival or cell number of osteoblasts.

• Korean Journal of Soil Science and Fertilizer
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• v.15 no.3
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• pp.166-171
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• 1982

A laboratiory experiment incubated at about $30^{\circ}C$ for 34 days was conducted in order to learn the effect of liming materials and rice straw on the volatilization of ammonia from flooded soils applied with urea. 1. The application of calcium hydroxide and calcium silicate increased buffer action of flood soil, though it resulted in increase in the volatilization of ammonia through raising flooded soil pH containing bicarbonate. 2. The mixing of rice straw powder to soil lowered pH of flooded soil, and decreased the volatilization of ammonia. The effect was particulary large when noliming material was used. 3. Calcium hydroxide depressed the evolution of $CO_2$ in the early days of incubation after flooding, while calcium silicate promoted the ammonification of soil nitrogen from the begining of flooding giving slow change in soil chemical properties. The rice straw was also effective in providing a favorable soil condition for the ammonification rather quickly.

• Korean Journal of Soil Science and Fertilizer
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• v.15 no.3
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• pp.162-165
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• 1982

As an approach to the explanation of increased availability of phosphate in reduced wetland soils, the interrelationships among pH, pe, $Fe^{+{+}}$ and water soluble phosphate in reduced soil-water suspension was studied. 1. p.e value of soil incubated for 8 weeks at $30^{\circ}C$ under waterlogged condition was sufficiently low to allow the conversion of strengite to vivianite. 2. The concentration of water soluble $Fe^{+{+}}$ in this system was higher than that is allowable by the solubility of vivianite. 3. From the relationship between pH and the concentration of water soluble $Fe^{+{+}}$, the concentration of water soluble $Fe^{+{+}}$ could be determined with the solubility of $FeCO_3$. 4. No definite relationship between pH and water soluble P was recognized which implied that the concentration of water soluble P in this system could not determined with the solubility of vivianite.