• Radiation Oncology Journal
• /
• v.19 no.2
• /
• pp.163-170
• /
• 2001

Purpose : The measurement of radiation survival using a clonogenic assay, the established standard, can be difficult and time consuming. In this study, We have used the MTT assay, based on the reduction of a tetrazolium salt to a purple formazan precipitate by living cells, as a substitution for clonogenic assay and have examined the optimal condition for performing this assay in determination of radiation sensitivity. Materials and Methods : Four human cancer cell lines - PCI-1, SNU-1066, NCI-H630 and RKO cells have been used. For each cell line, a clonogenic assay and a MTT assay using Premix WST-1 solution, which is one of the tetrazolium salts and does not require washing or solubilization of the precipitate were carried out after irradiation of 0, 2, 4, 6, 8, 10 Gy. For clonogenic assay, cells in $25\;cm^2$ flasks were irradiated after overnight incubation and the resultant colonies containing more than 50 cells were scored after culturing the cells for $10\~14$ days. For MTT assay, the relationship between absorbance and cell number, optimal seeding cell number, and optimal timing of assay was determined. Then, MTT assay was performed when the irradiated cells had regained exponential growth or when the non-irradiated cells had undergone four or more doubling times. Results : There was minimal variation in the values gained from these two methods with the standard deviation generally less than $5\%$, and there were no statistically significant differences between two methods according to t-test in low radiation dose (below 6 Gy). The regression analyses showed high linear correlation with the $R^2$ value of $0.975\~0.992$ between data from the two different methods. The optimal cell numbers for MTT assay were found to be dependent on plating efficiency of used cell line. Less than 300 cells/well were appropriate for cells with high plating efficiency (more than $30\%$). For cells with low plating efficiency (less than $30\%$), 500 cells/well or more were appropriate for assay. The optimal time for MTT assay was after 6 doubling times for the results compatible with those of clonogenic assay, at least after 4 doubling times was required for valid results. In consideration of practical limits of assay (12 days, in this study) cells with doubling time more than 3 days were inappropriate for application. Conclusion : In conclusion, it is found that MTT assay can successfully replace clonogenic assay of tested cancer cell lines after irradiation only if MTT assay was undertaken with optimal assay conditions that included plating efficiency of each cell line and doubling time at least.

• Korean journal of food and cookery science
• /
• v.16 no.6
• /
• pp.577-583
• /
• 2000

The purpose of this study was to investigate the effectiveness of various quality preservative treatments for extending shelf life and maintaining good quality of minimally processed fruit and vegetables produced in Korea. To determine the suitable treatments for delaying quality deterioration, fresh Asian pears and Chinese cabbages were sliced and treated with various quality preservatives (1% CaCl$_2$, 1% NaCl, 3% sucrose, 1% Ca-lactate, 1% vitamin C, 0.05% chitosan ＋1% vitamin C, 0.1% Sporix＋1% vitamin C, hot water (60$\^{C}$), 0.2% L-cysteine), packed with polyethylene film (60㎛-thick), and stored at 4$\^{C}$/0$\^{C}$ or 20$\^{C}$. Various biological and sensory tests were performed to evaluate the quality changes in minimally processed products. Results indicated that Chinese cabbages treated with 1% CaCl$_2$ at 4, and 1% CaCl$_2$ and 1% NaCl at 20$\^{C}$ were most effective in maintaining the quality and minimizing the biochemical changes during storage. For sliced Asian pears, 0.2% L-cysteine and 1% NaCl treatments were effective to reduce browning, and 1% CaCl$_2$ treatment was the most effective to prevent softening during storage at 20$\^{C}$ and 0$\^{C}$. Modified atmosphere packaging of Pleurotus ostreatus and Lentinus edodes had a significantly different shelf life depending on packaging material, packaging thickness and storage temperature. Sealed packaging with polyethylene film (60㎛-thick) for two kinds of mushrooms maintained a good quality with an extended shelf life by 30-50% at 20$\^{C}$ and by 30-130% at 0$\^{C}$. To minimize the quality deterioration which appeared in the condition of polyethylene film packaging, quality preservatives such as KMnO$_4$ and KHSO$_2$＋K$_2$S$_2$O$\_$5/ for SO$_2$ generation were added inside of mushroom packaging. The best condition for maintaining good quality longer was packaging with polyethylene film+SO$_2$ which showed 5080% extended shelf life for both Pleurotus ostreatus and Lentinus edodes at 20$\^{C}$ and 0$\^{C}$.

• Microbiology and Biotechnology Letters
• /
• v.36 no.3
• /
• pp.215-220
• /
• 2008

Rahnella aquatilis AY2000 has an unique characteristic which produces an anti-yeast substance (AYS). The AYS of the strain AY2000 was always secreted on agar plate, however, its activity in liquid culture was labile upon storage of the medium. In this paper, cultural conditions of the strain AY2000 for the AYS production were investigated in liquid culture, and minimal inhibitory concentration (MIC) against Saccharomyces cerevisiae was determined for the AYS activity. MIC of the AYS cultured in PYG broth at $^25{\circ}C$ for 24 hr was $23.5{\mu}g/mL$, however, that in MYCS (pH 5.5) broth at the same condition was $15.5{\mu}g/mL$. The activity of the AYS had increased rather in MYCS broth excluded $NH_4$-citrate than in the same broth contained $NH_4$-citrate, and MIC of the AYS produced in MYCS broth without $NH_4$-citrate was $15.5{\mu}g/mL$. When the strain AY2000 was maintained in MYCS broth without $NH_4$-citrate but added $100{\mu}M$ $FeCl_3$, the activity of the AYS had increased and its MIC was $7.8{\mu}g/mL$. MIC of the AYS was $7.8{\mu}g/mL$ after the strain AY2000 was cultured in MYCS broth containing $100{\mu}M$ $FeCl_3$ without $NH_4$-citrate, however, its MIC was $31.3{\mu}g/mL$ after 48-60 hr culture in the same broth.

• Archives of Reconstructive Microsurgery
• /
• v.5 no.1
• /
• pp.24-34
• /
• 1996

There are many kinds of free flaps for management of extensive soft tissue defect of extremities in orthopaedic field. Free vascularized scapular flap is one of the most useful and relatively easy to application. This flap has been utilize clinically from early eighties by many microsurgical pioneers. Authors performed 102 cases of this flap from 1984 to 1995. We have to consider about the surgical anatomy of the flap, technique of the donor harvesting procedures, vascular varieties and anatomical abnormalities and success rate and the weak points of the procedure. This flap nourished by cutaneous branches from circumflex scapular vessels emerges from the lateral aspect of the subscapular artery 2.5-5cm from its lateral origin passing through the triangular space(bounded by subscapularis, teres minor, teres major, long head of triceps). The terminal cutaneous branch runs posteriorly around the lateral border of the scapular and divided into two major branches, those transeverse horizontally and obliquely to the fascial plane of overlying skin of the scapular body. We can utilize these arteries for scapular and parascapular flap. The vascular pedicle ranged from 5 to 10 cm long depends on the dissection, usually two venae comitantes accompanied circumflex scapular artery and its major branches. The diameter of the circumflex scapular artery is more than 1mm in adult, rare vascular variation. Surgical techniques : The scapular flap can be dissected conveniently with prone or lateral decubitus position, prone position is more easier in my experience. There are two kinds of surgical approaches, most of the surgeon prefer elevation of the flap from its outer border towards its base which known easier and quicker, but I prefer elevation of the flap from its outer border because of the lowering the possibilities of damage to vasculature in the flap itself which runs just underneath the subcutaneous tissue of the flap and provide more quicker elevation of the flap with blunt finger dissection after secure pedicle dissection and confirmed the course from the base of the pedicle. There are minimal donor site morbidity with direct skin closure if the flap size is not so larger than 10cm width. This flap has versatility in the design of the flap shape and size, if we need more longer and larger one, we can use parascapular flap or both. Even more, the flap can be used with latissimus dorsi musculocutaneous flap and serratus anterior flap which have common vascular pedicle from subscapular artery, some instance can combined with osteocutaneous flap if we include the lateral border of the scapular bone or parts of the ribs with serratus anterior. The most important shortcoming of the scapular free flap is non sensating, there are no reasonable sensory nerves to the flap to anastomose with recipient site nerve. Results : Among our 102 cases, overall success rate was 89%, most of the causes of the failure was recipient site vascular problems such as damaged recipient arterial conditions, and there were two cases of vascular anomalies in our series. Patients ages from 3 years old to 62 years old. Six cases of combined flap with latissimus dorsi, 4 cases of osteocutaneous flap for bone reconstruction, 62 parascapular flap was performed - we prefer parascapular flap to scapular. Statistical analysis of the size of the flap has less meaningful because of the flap has great versatility in size. In the length of the pedicle depends on the recipient site condition, we can adjust the pedicle length. The longest vascular pedicle was 14 cm in length from the axillary artery to the enter point cutaneous tissue. In conclusion, scapular free flap is one of the most useful modalities to manage the large intractable soft tissue defect. It has almost constant vascular pedicle with rare anatomical variation, easy to dissect great versatility in size and shape, low donor morbidity, thin and hairless skin.

• Maxillofacial Plastic and Reconstructive Surgery
• /
• v.31 no.4
• /
• pp.281-286
• /
• 2009

Introduction: Enamel matrix derivative (EMD) is a protein which is secreted by Hertwig root sheath and plays a major role in the formation of cementum and attachment of peridontium. Several studies have shown that EMD promoted the proliferation and differentiation of preosteoblasts, osteoblasts and periodontal ligament cells in vitro: however, reports showing the inhibition of osteogenic differentiation by EMD also existed. This study was designed to simultaneously evaluate the effect of EMD on the two cell lines (human mesenchymal stem cells: hMSC, human periodontal ligament derived fibroblasts: hPDLCs) by means of quantitative analysis of some bone related matrices (Alkaline phosphatase : ALP, osteopontin ; OPN, osteocalcin ; OC). Materials and Methods: hMSCs and hPDLCs were expanded and cells in the 4${\sim}$6 passages were adopted to use. hMSc and hPDLCs were cultured during 1,2,7, and 14 days with 0, 50 and 100 ${\mu}g/ml$ of EMD, respectively. ALP activity was assessed by SensoLyte ALP kit and expressed as values of the relative optical density. Among the matrix proteins of the bony tissue, OC and OPN were assessed and quantification of these proteins was evaluated by means of human OC immunoassay kit and human OPN assay kit, respectively. Results: ALP activity maintained without EMD at $1,2^{nd}$ day. The activity increased at $7^{th}$ day but decreased at $14^{th}$ day. EMD increased the activity at $14^{th}$ day in the hPDLCs culture. In the hMSCs, rapid decrease was noted in $7^{th}$ and $14^{th}$ days without regard to EMD concentrations. Regarding the OPN synthesis in hPDLCs, marked decrease of OPN was noted after EMD application. Gradual decrease tendency of OPN was shown over time. In hMSCs, marked decrease of OPN was also noted after EMD application. Overall concentration of OPN was relatively consistent over time than that in hPDLCs. Regarding the OC synthesis, in both of hPDLCs and hMSCs, inhibition of OC formation was noted after EMD application in the early stages but EMD exerted minimal effect at the later stages. Conclusion: In this experimental condition, EMD seemed to play an inhibitory role during the differentiation of hMSCs and hPDLCs in the context of OC and OPN formation. In the periodontium, there are many kinds of cells contributing to the regeneration of oral tissue. EMD enhanced ALP activity in hPDLCs rather than in hMSCs and this may imply that EMD has a positive effect on the differentiation of cementoblasts compared with the effect on hMSCs. The result of our research was consistent with recent studies in which the authors showed the inhibitory effect of EMD in terms of the differentiation of mineral colony forming cells in vitro. This in vitro study may not stand for all the charateristics of EMD; thus, further studies involving many other bone matrices and cellular attachment will be necessary.

• The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
• /
• v.17 no.2
• /
• pp.45-58
• /
• 2012

For the development of reference values and evaluation of water quality in various environmental conditions, we divided the coastal region around Korean peninsular into 5 distinctive ecological regions based on the influence of surface current, depth, tidal range, turbidity, and climate condition. We used national marine environment monitoring data collected by National Fisheries Research & Development Institute(NFRDI) from 2000-2009. For the reference values, we used maximum seasonal mean from 2000 to 2007 for DIN, DIP, and chlorophyll-a and minimum seasonal mean for secchi depth measured at stations without the influence of river runoff in each ecological regions. For the reference value of bottom dissolved oxygen saturation, we used minimum mean value of 90% calculated from minimal riverine influence stations of whole regions. We calculated enrichment score for each assessment criteria. The enrichment score of DIN, DIP, and Chlorophyll-a was 1 (=< reference value), 2 (< 110% of reference value), 3 (< 125% of reference value), 4 (< 150% of reference value), and 5 (> 150% of reference value). The enrichment score of DO saturation and Secchi depth was 1 (> reference value), 2 (> 90% of reference value), 3 (>75 % of reference value), 4 (> 50% of reference value), and 5 (< 50% of reference value). We calculated water quality index using weighted linear combination of five enrichment score for the comparison of whole regions. From the water quality index distribution calculated from all stations between 2000 and 2007 period, we classified into 5 grade based on the standard deviation calculated from total water quality index. We assigned grade very good(I), good(II), moderate(III), bad(IV), and very bad(V) when the water quality index was less than 23, minimum + 1 sd, +2 sd, +3 sd, and grater than minium+ 3 sd, respectively.

• Journal of the korean academy of Pediatric Dentistry
• /
• v.34 no.1
• /
• pp.36-42
• /
• 2007

To compare the survival rate of periodontal ligament cells preserved in storage media with good availability at the time of an avulsion injury, periodontal ligament cells were incubated in ${\alpha}-MEM$ culture medium containing 10% FBS in condition of $37^{\circ}C$, 5% $CO_2$. These cells were then cultured in HBSS, ${\alpha}-MEM$, milk(S co., P. co.) and tap water at the temperature of 4, 25, $37^{\circ}C$ each in 60 min. The groups were measured by MTT assay. The results were as follows : 1. Among the storage media at $4^{\circ}C$, ${\alpha}-MEM$ and P-milk had the highest preserving ability of periodontal ligament cells, while that of HBSS S-milk and tap was low in order. 2. Among the storage media at $25^{\circ}C$, ${\alpha}-MEM$ had the highest preserving ability of periodontal ligament cells, while that of P-milk, HBSS, S-milk, tap water was low in order. 3. Among the storage media at $37^{\circ}C$, the preserving ability of periodontal ligament cells was very high in ${\alpha}-MEM$, P-milk, HBSS and S-milk, it's lowest in tap water. 4. The preserving ability of periodontal ligament cells in ${\alpha}-MEM$ was high at $4^{\circ}C$ and it's low in order of $25^{\circ}C$, $37^{\circ}C$, but in HBSS was high at $4^{\circ}C$ and it's low at $25^{\circ}C$, $37^{\circ}C$ 5. The preserving ability of periodontal ligament cells in S-milk and P-milk was high at $4^{\circ}C$, $25^{\circ}C$ and it s low at $37^{\circ}C$. In conclusion, HBSS is the storage medium of choice in an avulsion, but in this study it is preferable to choose milk at $4^{\circ}C$ for tooth since it is easy to get and affect cell viability.

• Food Science and Preservation
• /
• v.11 no.2
• /
• pp.195-200
• /
• 2004

Microbial lethal value and nutrient retention of sous vide processed spinach were evaluated with mathematical model prediction and experimental trial for different package sizes and pasteurization temperatures. The package size covers 500 g, 1 kg and 2 kg, while the pasteurization temperature includes 80, 90 and 97$^{\circ}C$. The basic process scheme consists of filling blanched spinach into barrier plastic film pouch, sealing under vacuum, pasteurization in hot water with over pressure and final cooling to 3$^{\circ}C$. Pasteurization condition was designed based on attainment of 6 decimal inactivation of Listeria monocytogenes at geometric center of the pouch package by heating cycle, which was determined by general method. Heat penetration property of the package and thermal destruction kinetics were combined to estimate the retention of ascorbic acid and chlorophyll. Smaller packages with shorter pasteurization time gave better nutrient retention, physical and chemical qualities. Larger package size was estimated and confirmed experimentally to give higher pasteurization value at center, lower ascorbic acid and chlorophyll contents caused by longer heat process time. Lower pasteurization temperature with longer process time was predicted to give lower pasteurization value at center and lower ascorbic acid, while chlorophyll content was affected little by the temperature. Experimental trial showed better retention of ascorbic acid and chlorophyll for smaller package and higher pasteurization temperature with shorter heating time. The beneficial effect of smaller package and higher pasteurization temperature was also observed in texture, color retention and drip production.

• Journal of Korean Society of Environmental Engineers
• /
• v.36 no.10
• /
• pp.719-723
• /
• 2014

For effective implementation of total maximum daily load (TMDL), this study presented the improving plans of non-point source pollution management including the classification of non-point source pollution, calculation of non-point source pollution load (generated, discharged), selection of non-point source pollution management regions and management of non-point source pollutant. First of all, the definition of point source pollution and non-point source pollution based on the legal and scientific viewpoint should be precisely classified and managed. Especially, the forest, grassland and river without occurrence of environmental damage by activity of business and human should be separately classified natural background pollutants. The unit for generated and discharged non-point source pollution should be preferentially changed according to actual condition of watershed. The calculation methods of generated and discharged non-point source pollution should be corrected consideration on the amount and duration of rainfall. While the TMDL is implemented, non-point source pollution management regions should be selected in the watersheds exceed the targeted water quality standards by the rainfall. The non-point source pollution management regions should be selected in the minimal regions where have high values of discharged non-point source pollution density in the urban area, farmland and site area except forest, grassland in the whole watershed. The non-point source pollutant treatment facilities, which take into consideration non-point source pollution load per unit area, duration of the excess concentration, realizable possibility of treatment, effectiveness of treatment cost versus point source pollutant, should be established in the regions with a large generated non-point source pollution load and a high concentration of water quality exceed the targeted water quality standards by the rainfall.

• Journal of Life Science
• /
• v.16 no.1
• /
• pp.131-135
• /
• 2006

Oligopeptide is known to be an essential nitrogen nutrient for bacterial growth. Oligopeptide can be transported into cytoplasm by a specific transport system, Opp system. Opp system is composed of five proteins, which are transcribed by an operon. These are responsible for oligopeptide binding protein (OppA), permease (OppB and OppC) and energy generation system (OppD and OppF), respectively. Previously, we isolated the opp operon from Vibrio fluvialis and constructed the oppA mutant by allelic exchange method. In this study, we investigated the growth pattern and biofilm production under the different growth condition. When the cells were cultivated using brain heart infusion(BHI) medium, the wild type was faster than the mutant in growth during the exponential phase. However, it showed that the growth pattern of two strains in M9 medium is very similar. The growth of wild type showed better than that of the mutant grown at pH 8. At pH 7, there was no an obvious difference in growth. After 5 mM $H_2O_2$ was treated to the cells $(OD_{600}=1.2)$, the cell survival was examined. The oppA mutation did not affect in survivability. In the presence of $10{\mu}g/ml$ polymyxin B, the biofilm production of the oppA mutant was higher than that of the wild type.