• Journal of Life Science
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• v.10 no.4
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• pp.408-421
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• 2000

We tried to establish the culture method of the chondrocyte isolated from the epiphyseal cartilage and to investigate morphological changes of chondrocyte cultured with enzyme-digested costal cartilage, the perichondrium and experimentally damaged meniscus of rabbit. De novo chondrocyte pellets were prepared from epiphyseal plates by culturing isolated epiphyseal chondrocytes from 4 week. old rabbits. We morphologically assessed the cartilage formation of the chondrocyte culture with enzyme-digested costal carilage, the perichondrial culture, the cultured chondrocytes transplants into experimentally damaged meniscus of rabbits, the perichondrial culture, the cultured chondrocytes transplants into experimentally damaged meniscus of rabbit. In the 24 days, the epiphyseal chondrocytes maintained the typical phenotypes of the partial nodular cell formation. The 30 days cryopreserved chondrocytes showed abnormal and irregular shape. In the type II collagen added culture, the chondrocytes showed expanded rough endoplasmic reticulum and small and large round-like vesicles of processes. In the type IV collagen added culture, the chondrocytes showed large perinuclear vaculoes and abundant well-developed rough endoplasmic reticulum of processes. In the culture with enzyme- digested costal cartilage and the perichondrial culture, the chondrocytes showed a few swelling rough endoplasmic reticulum and vacuoles. The cultured epiphyseal chondrocytes maintained typical phenotype and the chondrocytes were grown faster and maintained more typical phenotype in the type II and IV collagen added culture. The transformed chondrocytes secreted abundant extracellular matrix in the type II collagen added culture, and showed processes in the type IV collagen added culture. The perichondrial chondrocytes were grown faster and their implants were able to transplant. The cultured chondrocytes transplanted into experimentally damaged meniscus were adapted between the meniscus tissues. And the immunocyto-chemical reaction of the type II collagen of the chondrocytes were found to be maintained. The chondrocytes cultured cartilage. The chondrocytes secreted abundantly. The cultured chondrocytes transplanted into experimentally damaged meniscus changed immature cells into enlarged mature cells with extracellular secretion.

• Journal of the Korean Society of Food Culture
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• v.24 no.5
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• pp.533-539
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• 2009

Frying is the most popular cooking style used in food service institutions in Korea because fried dishes are well accepted by students. However, trans fat contents reduction have recently been required in many foods for health reasons Therefore, alternatives to frying such as oven baking or roasting are being used in many institutions. Steam convection ovens provide dry heat as well as steam so that they can be used to make a wide variety of Korean dishes. In this study, a menu acceptance test was conducted over 2 weeks. Pork, chicken, fish and potato dishes cooked by conventional cooking methods were served for 4 successive days, after which the same dishes were prepared using the oven. Overall, 322 junior high school students evaluated the traditionally cooked foods, while 316 evaluated the oven cooked foods. Comparison of the foods prepared using both methods only revealed a significant difference in the acceptability of foods on the fish menu (p<0.05). Specifically, the acceptance of fried fish was higher than that of the oven baked fish. Additionally, overall acceptance of the menu by males was higher than the acceptance by girls. Furthermore, students who had the preference for special ingredients showed a higher menu acceptance for the menu cooked with those ingredients. On average, approximately 25% of the meal was not consumed and left as plate waste. The portion of the fried fish not consumed was smaller than that of the oven cooked fish, but the portion not consumed did not vary based on cooking method for any other foods evaluated. Overall, it is expected that the oven cooking method will be a good substitute for frying or other cooking method for traditional Korean dishes.

• Journal of Microbiology
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• v.33 no.2
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• pp.146-148
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• 1995

Coprinus congregatus has a membrane-associated laccase which is not secreted into culture media. A mutant monokaryon obtained, by U. V. irradiation followed by protoplast generation and regeneration method, was successfully isolated. When the mutant was grown on a agar plate or in a liquid medium, it secreted laccase while the wild type did not under the same growth conditions. The laccase of the mutant was compared with that of wild type did not under the same growth conditions. The laccase of the mutant was compared with that of wild type of native PAGE analysis, and showed identical mobility.

• Journal of Microbiology and Biotechnology
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• v.24 no.11
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• pp.1559-1565
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• 2014

Cellulase and xylanase are main hydrolysis enzymes for the degradation of cellulosic and hemicellulosic biomass, respectively. In this study, our aim was to develop and test the efficacy of a rapid, high-throughput method to screen hydrolytic-enzyme-producing microbes. To accomplish this, we modified the 3,5-dinitrosalicylic acid (DNS) method for microwell plate-based screening. Targeted microbial samples were initially cultured on agar plates with both cellulose and xylan as substrates. Then, isolated colonies were subcultured in broth media containing yeast extract and either cellulose or xylan. The supernatants of the culture broth were tested with our modified DNS screening method in a 96-microwell plate, with a $200{\mu}l$ total reaction volume. In addition, the stability and reliability of glucose and xylose standards, which were used to determine the enzymatic activity, were studied at $100^{\circ}C$ for different time intervals in a dry oven. It was concluded that the minimum incubation time required for stable color development of the standard solution is 20 min. With this technique, we successfully screened 21 and 31 cellulase- and xylanase-producing strains, respectively, in a single experimental trial. Among the identified strains, 19 showed both cellulose and xylan hydrolyzing activities. These microbes can be applied to bioethanol production from cellulosic and hemicellulosic biomass.

• The Korean Journal of Malacology
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• v.29 no.3
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• pp.197-205
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• 2013

Even though seedling production of Haliotis discus hannai has fully developed, the culture of benthic diatom as a live food for larvae is still a barrier to solve in commercial hatchery. The farmer depends on mixed microalgae which are non-selectively attached on the plate by flowing of natural seawater. The adequate diatom on the plate for the larvae in terms of quality and quantity is always significant bottleneck in the hatchery. In this study, two benthic diatom species, Caloneis schroederi and Rhaphoneis sp. were separately cultured in mass and inoculated to four tons' settlement tank of the larvae. And the larvae and the spats were cultured for nineteen and nine weeks, respectively. The result on seedling production of H. discus hannai with this method was compared to that of the farmer's traditional method as a control. With regard to variation of species composition of benthic diatom on the plate, C. schroederi and Rhaphoneis sp. were dominant for first three weeks after inoculation. But the diverse diatoms mainly, Navicula, Amphora, Cylindrotheca, Licmophora, Pleurosigma began to attache on the plate from the 4th week. The larvae attached 2.5 times more in C. schroederi tank than in the control tank. The final total biomass of the seeds in Rhaphoneis sp. tank was 3.2 times more than that of the control tank. The retared-spats in the seedling production also showed significantly higher growth and survival in the spat fed Rhaphoneis sp. or C. schroederi than those in the control group. We suggest that C. schroederi is proper for settlement of the larvae and Rhaphoneis sp. is appropriate for the growth of the larvae and spats in the commercial hatchery of H. discus hannai.

• The Journal of the Korean Society for Microbiology
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• v.19 no.1
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• pp.41-48
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• 1984

The present study was undertaken to elucidate the antibacterial spectrum of L. casei phage type $J_1$ strain isolated from a fermented milk product against pathogenic enteric bacteria. Growth inhibitory effects and minimum inhibitory concentration(MIC) of culture supernatants of L. casei grown in MRS broth were measured by both plate culture method and microplate broth dilution technique against Salmonella typhi, Salmonella typhimurium, Shigella flexneri, Shigella dysenteriae, enterpathogenic E. coli, Klebsiella pneumoniae and Pseudomonas aeruginosa. The results are summarized as follows: 1. The MRS broth culture of L. casei gave a similar extent of growth inhibitory effects against S. typhi, S. typhimurium, S. flexneri, S. dysenteriae, E. coli, K. pneumoniae and P. aeruginosa, respectively. 2. The inhibitory effects of L. casei culture were observed either in whole broth culture or in culture supernatant, but neither the bacterial suspension nor the neutralized culture supernatant showed such as antibacterial activities. 3. The MIC titres of the culture supernatants were ${\log_2}5$ to ${\log_2}6$, whereas those of the neutralized culture supernatant dropped markdely to ${\log_2}2$ to ${\log_2}3$. These results indicated that major portion of growth inhibitory effects of MRS broth culture of L. casei against enteric bacterial pathogens was possibly due to the acids produced, and minor portion to other antibacterial substances.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.32 no.3
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• pp.272-278
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• 2006

Purpose: This study is designed to evaluate biocompatibility of three types of absorbable collagen GBR membrane in vitro. Material and Method: The human PDL fibroblasts culture was obtained through typical way and the cells used in the experiment was forth passage. The membranes examined were Experimental group A, B, C. All the 3-experimental groups were made of bovine pericardium and the membranes were excised into 5$\times$5mm respectively. The samples of the membranes were fixed on the 24-well plate with the double-sided adhesive tape. Then, 2ml of cell suspension which included $2{\times}10^4$cells was inoculated into the 24-well plate, and the cells were cultured for 1 week. Cellular viability and the alkaline phosphatase activity were measured with ELISA. The membranes in the culture were processed to examine with SEM. Results: The survival rate was highest in control and Experimental group A is the next, group B and group C in order of the value. The values are analyzed for statistical difference using Wilcoxon test. All the values of experimental groups are significantly lower than those of control, and the vaules among the experimental groups significantly differ from each other. Alkaline phosphatase level was identical order with the viable cell rate. SEM examination revealed that the PDL fibroblasts adherent on culture dish (control) and group A were spindle-shaped, but on group B and C, the cells were round-shaped without processes.

• Journal of Microbiology and Biotechnology
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• v.27 no.1
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• pp.101-111
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• 2017

The task of improving a fungal strain is highly time-consuming due to the requirement of a large number of flasks in order to obtain a library with enough diversity. In addition, fermentations (particularly those for fungal cells) are typically performed in high-volume (100-250 ml) shake-flasks. In this study, for large and rapid screening of itaconic acid (IA) high-yielding mutants of Aspergillus terreus, a miniaturized culture method was developed using 12-well and 24-well microtiter plates (MTPs, working volume = 1-2 ml). These miniaturized MTP fermentations were successful, only when highly filamentous forms were induced in the growth cultures. Under these conditions, loose-pelleted morphologies of optimum sizes (less than 0.5 mm in diameter) were casually induced in the MTP production cultures, which turned out to be the prerequisite for the active IA biosynthesis by the mutated strains in the miniaturized fermentations. Another crucial factor for successful MTP fermentation was to supply an optimal amount of dissolved oxygen into the fermentation broth through increasing the agitation speed (240 rpm) and reducing the working volume (1 ml) of each 24-well microtiter plate. Notably, almost identical fermentation physiologies resulted in the 250 ml shake-flasks, as well as in the 12-well and 24-well MTP cultures conducted under the respective optimum conditions, as expressed in terms of the distribution of IA productivity of each mutant. These results reveal that MTP cultures could be considered as viable alternatives for the labor-intensive shake-flask fermentations even for filamentous fungal cells, leading to the rapid development of IA high-yield mutant strains.

• Archives of Pharmacal Research
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• v.21 no.3
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• pp.253-259
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• 1998

In this study, we developed immunoassay methods for the more convenient and effective detection of rat tracheal mucin and the results were compared with those of [$3^H$]glucosamine based gel-filtratioh method. A monoclonal anti-rat tracheal mucin antibody, mAbRT03, which specifically recognizes rat tracheal mucins, was used throughout in this study. To induce mucin secretion, varying concentrations of ATP (0-2 mM) were applied to the primary rat tracheal surface epithelial (RTSE) cell culture which had been metabolically radiolabeled with [$3^H$]glucosamine and the secretion of mucin was analyzed both by the immunoassay and the gel-filtration chromatography methods. For the immunoassay, the following two procedures were employed. 1) Simple ELISA; the culture spent media were directly coated onto the assay plate and the immunoreactivity with mAbRT03 was assessed from the standard curve generated with the purified rat mucin. 2) Inhibition ELISA; A known amount of the purified rat mucin was coated onto the assay plate and then ATP-stimulated culture spent media were added to inhibit the immunorelitivity with mAbRT03. The contents of mucin in the sample were calculated from the standard inhibition curve which was generated with the purified rat mucin. The assay results obtained from the immunoassays were identical with those from the gel-filtration methods. The present result indicates that ELISA can be substituted for the laborious, time-consuming gel-filtration assay in studying the regulation of airway mucin release from cultured airway epithelial cells.

• Journal of Food Hygiene and Safety
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• v.26 no.4
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• pp.361-365
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• 2011

To evaluate the indoor air quality of food manufacturing plants, the presence of viable bacteria and fungi was assessed in the indoor air of the facilities at which 9 food items were manufactured. Air samples were collected from the general zone, low clean zone and clean zone of each factory with an air sampler, in combination with plate counts agar using for bacteria, and dichloran-glycerol agar for fungi. The samples were incubated at $25^{\circ}C$ for 4 to 7 days. After culture, the colony forming units (CFU) on each plate were counted and corrected with a positive hole conversion table. The average concentration of bacteria was $2.2{\times}10^3\;CFU/m^3$ in the general zone, $1.2{\times}10^3\;CFU/m^3$ in the low clean zone and $7.3{\times}10^2\;CFU/m^3$ in the clean zone. The average concentration of fungal microbes was $2.5{\times}10^3\;CFU/m^3$ in the general zone, $2.6{\times}10^3\;CFU/m^3$ in the low clean zone, and $2.0{\times}10^2\;CFU/m^3$ in the clean zone. No meaningful differences were detected between the general zone and the low clean zone, but the clean zone had significantly lower concentrations than the other zones. Additionally, the identification of the fungi was performed according to morphological method using a giant culture and slide culture. The fungi were identified as belonging to 18 genera, and the genera Cladosporium(33%), Penicillium(29%) and Aspergillus(26%), predominated. Aspergillus isolates were identified to species level, and A. ochraceus, a mycotoxigenic species, was identified. As part of the effort to control the quality of the indoor air of food manufacturing plants, our results show that continued studies are clearly warranted.