• Title/Summary/Keyword: Primary culture

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Development of a predictive model describing the growth of Staphylococcus aureus in processed meat product galbitang (식육추출가공품 중 갈비탕에서의 Staphylococcus aureus 성장예측모델 개발)

  • Son, Na-Ry;Kim, An-Na;Choi, Won-Seok;Yoon, Sang-Hyun;Suh, Soo-Hwan;Joo, In-Sun;Kim, Soon-Han;Kwak, Hyo-Sun;Cho, Joon-Il
    • Korean Journal of Food Science and Technology
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    • v.49 no.3
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    • pp.274-278
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    • 2017
  • In this study, predictive mathematical models were developed to estimate the kinetics of Staphylococcus aureus growth in processed meat product galbitang. Processed meat product galbitang was inoculated with 0.1 mL of S. aureus culture and stored at 4, 10, 20, $37^{\circ}C$. The ${\mu}_{max}$ (maximum specific growth rate) and LPD (lag phase duration) values were calculated. The primary model was used to develop a response surface secondary model. The growth parameters were analyzed using the square root model as a function of storage temperature. The developed model was confirmed by calculating RMSE (Root Mean Square Error) values as statistic parameters. The LPD decreased, but ${\mu}_{max}$ increased with an increase in the storage temperature. At 4, 10, 20 and $37^{\circ}C$, $R^2$ was 0.99, 0.98, 0.99 and 0.99, respectively; RMSE was 0.39. The developed predictive growth model can be used to predict the risk of S. aureus contamination in processed meat product galbitang; hence, it has potential as an input model for the risk assessment.

Isolation and Characterization of the IAA Producing Methylotrophic Bacteria from Phyllosphere of Rice Cultivars(Oryza sativa L.) (벼(Oryza sativa L.)의 잎 면으로부터의 IAA를 생성하는 Methylotrophic Bacteria의 분리 선별 및 특성 비교)

  • Lee, Kyu-Hoi;Munusamy , Madhaiyan;Kim, Chung-Woo;Lee, Hyoung-Seok;Selvaraj, Poonguzhali;Sa, TongMin
    • Korean Journal of Soil Science and Fertilizer
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    • v.37 no.4
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    • pp.235-244
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    • 2004
  • In this study, we compared the levels of methylotrophic bacterial community diversity in the leaf samples of 19 rice cultivars collected from three regions of Korea. Nineteen pink pigmented isolates showing characteristic growth on methanol were obtained. Physiological and biochemical characters of each isolate were examined according to methods described in Bergey's Manual of Systematic Bacteriology. When phylotypes were defined by performing numerical analysis of 37 characteristics, four distinct clusters were formed. The two reference strains, Methylobacterium extorquens AM1 and Methylobacterium fujisawaense KACC10744 were found to group under cluster IV and cluster III respectively. Cluster I diverged on the basis of nitrate reduction and four isolates showed tolerance upto 0.5 M NaCl concentrations. Two strains in cluster I and III were found to possess methane utilizing properties. Most of the isolates in all the four clusters utilized monosaccharides, disaccharide and polyols as carbon source. When the isolates were subjected for indole-3-acetic acid (IAA) analysis in the presence of L-tryptophan, only 8 isolates exhibited IAA production. In addition, the nitrogen source in the medium was found to influence the IAA production. Addition of $(NH_4)_2SO_4$ in the medium led to a 2 to 30 fold increase in the indole synthesis. However, $KNO_3$, $NH_4NO_3$ and $NH_4Cl$ substitution did not significantly stimulate the synthesis of IAA in the growth medium. Result of gnotobiotic root elongation assay significantly increased roots and shoots lengths, and number of lateral roots, which is mediated by IAA production in the culture medium. The rice seedlings primary roots from seeds treated with methylotrophic isolates were on average 27 to 56% longer than the roots from seeds treated with the uninoculated seeds. In addition, application of different high concentrations of authentic IAA ($400g\;mL^{-1}$) to roots of rice seedlings inhibited root growth. However, the IAA concentration from 10 to $200g\;mL^{-1}$, IAA promoted root growth of rice seedlings. These results suggest that bacterial IAA plays a major role in the development of the host plant root system.

Anti-obesity effect of EGCG and glucosamine-6-phosphate through decreased expression of genes related to adipogenesis and cell cycle arrest in 3T3-L1 adipocytes (Adipogenesis관련 유전자발현감소와 Cell Cycle Arrest를 통한 EGCG와 Glucosamine-6-Phosphate의 Anti-Obesity 효과)

  • Kim, Kkot Byeol;Jang, Seong hee
    • Journal of Nutrition and Health
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    • v.47 no.1
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    • pp.1-11
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    • 2014
  • Purpose: Several studies have proven that EGCG, the primary green tea catechin, and glucosamine-6-phosphate (PGlc) reduce triglyceride contents in 3T3-L1 adipocytes. The objective of this study is to evaluate the combination effect of EGCG and PGlc on decline of accumulated fat in differentiated 3T3-L1 adipocytes. Methods: EGCG and PGlc were administered for 6 day for differentiation of 3T3-L1 adipocytes. Cell viability was measured using the CCK assay kit. In addition, TG accumulation in culture 3T3-L1 adipocytes was investigated by Oil Red O staining. We examined the expres-sion level of several genes and proteins associated with adipogenesis and lipolysis using real-time RT-PCR and Western blot analysis. A flow cytometer Calibar was used to assess the effect of EGCG and PGluco on cell-cycle progression of differentiating 3T3-L1 cells. Results: Intracelluar lipid accumulation was significantly decreased by combination treatment with EGCG $60{\mu}M$ and PGlc $200{\mu}g/m$ compared with control and EGCG treatment alone. In addition, use of combination treatment resulted in directly decreased expression of $PPAR{\gamma}$, $C/EBP{\alpha}$, and SREBP1. In addition, it inhibited adipocyte differentiation and adipogenesis through downstream regulation of adipogenic target genes such as FAS, ACSL1, and LPL, and the inhibitory action of EGCG and PGlc was found to inhibit the mitotic clonal expansion (MCE) process as evidenced by impaired cell cycle entry into S phase and the S to G2/M phase transition of confluent cells and levels of cell cycle regulating proteins such as cyclin A and CDK2. Conclusion: Combination treatment of EGCG and PGlc inhibited adipocyte differentiation through decreased expression of genes related to adipogenesis and adipogenic and cell cycle arrest in early stage of adipocyte differentiation.

The application of alternative methods for skin irritation evaluation on pesticides (농약에 대한 피부자극성 대체시험법 적용연구)

  • Jeong, Mi-Hye;Kim, Mi-Kyoung;Park, Soo-Jin;You, Are-Sun;Hong, Soon-Sung;Park, Kyung-Hun;Park, Jae-Eup
    • The Korean Journal of Pesticide Science
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    • v.16 no.3
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    • pp.261-266
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    • 2012
  • It is common to use many experiment animals to evaluate the toxicity of chemicals including pesticides. For protecting animal, the concepts of 3R (Reduction, Replacement, Refinement) were introduced and in vitro alternatives methods actively have been developed all over the world. Many experimental animals for toxicological tests have been used, so that it is important to establish the alternative methods. In this study, the alternative method using reconstituted human skin model (Keraskin$^{TM}$) was conducted for classification of skin irritation on pesticides. Sixteen formulations selected on the basis of the degree of irritation were treated by Keraskin$^{TM}$ test. The percent of cell viability was measured into the culture medium collected after treatment of the pesticides for 24-72 hrs. The skin irritations of formulations were evaluated by the cell viability. In this study, The 4 formulations with mild irritation in rabbits were evaluated as nonirritant, the 6 formulations with moderate and severe irritation were evaluated as irritant in human skin model test. We suggest that the alternative test using Keraskin$^{TM}$ model could be used as toxicity evaluation for primary irritation index (P.I.I.) score of greater than or equal to 2.1 of pesticides. The further studies should be required to apply for hazardous assessment of pesticides on alternative skin irritation methods because of the interindividual variability of the sensitivity of skin irritation on pesticides.

Construction of Retrovirus Vector System for the Regulation of Recombinant hTPO Gene Expression (재조합 hTPO 유전자의 발현 조절을 위한 Retrovirus Vector System의 구축)

  • Kwon, Mo-Sun;Koo, Bon-Chul;Kim, Do-Hyang;Kim, Te-Oan
    • Reproductive and Developmental Biology
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    • v.31 no.3
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    • pp.161-167
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    • 2007
  • In this study, we constructed and tested retrovirus vectors designed to express the human thrombopoietin gene under the control of the tetracycline-inducible promoters. To increase the hTPO gene expression at him-on state, WPRE sequence was also introduced into retrovirus vector at downstream region of either the hTPO gene or the sequence encoding reverse tetracycline-controlled transactivator (rtTA). Primary culture cells (PFF, porcine fetal fibroblast; CEF, chicken embryonic fibroblast) infected with the recombinant retrovirus were cultured in the medium supplemented with or without doxycycline for 48hr, and induction efficiency was measured by comparing the hTPO gene expression level using RT-PCR, western blot and ELISA. Higher hPTO expression and tighter expression control were observed from the vector in which the WPRE sequence was placed at downstream of the hTPO (in CEF) or rtTA(in PFF) gene. This resulting tetracycline inducible vector system may be helpful in solving serious physiological disturbance problems which have been a major obstacle in successful production of transgenic animals.

Increased Protein of the Secretory Leukocyte Pretense Inhibitor (SLPI) and the Expression of Growth Factors in NIH3T3 Cells by LPS Stimulation (NIH3T3 세포주에서 LPS자극에 의한 분비백혈구단백분해효소억제제 (SLPI)의 단백질증가와 성장인자들의 발현)

  • Lee, Sang-Hwa;Choi, Baik-Dong;Jeong, Soon-Jeong;Jang, Hyun-Seon;Kim, Byung-Ock;Lim, Do-Seon;Park, Joo-Cheol;Wang, Guan-Lin;Jeong, Moon-Jin
    • Applied Microscopy
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    • v.36 no.3
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    • pp.165-172
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    • 2006
  • Secretory leukocyte protease inhibitor (SLPI) involves tissue protection against the destructive action of neutrophil elastase at the site of inflammation. Several studies on new functions of SLPI have demonstrated that SLPI may play a primary role in innate immunity than protease inhibitor, To identify the function of SLPI by lipopolysaccharide (LPS) stimulation in the embryonic fibroblast (NIH3T3) cells. we studied the expression of SLPI compared to other growth factors involving the LPS treatment. To address this, we performed the reverse transcriptase polymerase chain reaction (RT-PCR) and Western blots for the detection of mRNA and protein expression of the SLPI and some growth factors such as VEGF. bFGF, and PDGF-BB after LPS stimulation. NIH3T3 cells were exposed 100 ng/mL Escherichia coli LPS for 30min, 60min, 90min, 24h, and 48h, respectively. The result of RT-PCR showed that SLPI and VEGF mRNA was expressed strongly in NIH3T3 without related to LPS stimulation. mRNA of bFGF was weakly expressed such as the expression of the control. PDGF mRNA expression gradually increased follows at time course. However, SLPI protein level was increased in lysates and culture medium by LPS stimulation. Phase contrast microscopic and scanning electron microscopic observation showed that the increased cell number and cytoplasmic enlargement of the NIH3T3 cells. Therefore, it suggests that the LPS upregulates SLPI expression in NIH3T3 cells. Moreover, secreted SLPI may stimulate cell proliferation and migration.

A Study on the Reproduction Experimental of Breaking of dried stalks technique of Recorded in Oju-Yeonmunjang jeon-Sango in the late Joseon Dynasty (조선 후기 『오주연문장전산고』에 기록된 자연건조쇄경식 제섬 기술 재현 실험 연구)

  • Kong, Sanghui;Ree, Jiwon;Kim, Hajin
    • Korean Journal of Heritage: History & Science
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    • v.52 no.4
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    • pp.170-183
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    • 2019
  • Fiber scutching refers to the process of extracting fibers from plants by separating or extracting fibers from the raw materials. As the definition of the term implies, the "Fiber Scutching" is performed on plants with advanced bast fiber as the primary material processing technique performed on plant materials. Some of the most popular phosphorus plants are ramie, hemp, flax, and the paper mulberry, which have a long history of cultivation and a wide range of distribution, making them very universal as a material supporting human life and culture. This study was described in Oju-yeonmunjangjeon-sango but was designed to re-examine the method of breaking dried stalks, which is currently unused in Korea, to examine the feasibility and characteristics of the technology. As a result of sampling and experimenting with hemp bast using the method recorded in the literature, hemp fiber was actually produced. The criteria for removing the shell from the hemp stem were the degree of discoloration and drying, and only when the stalk was completely discolored to yellow could segregation of the stalk from the shell be performed. The amount of sunlight and temperature were conditions that accelerated drying. However, if exposed for a long time, it is confirmed that hemp bast will be in a suitable condition to process, regardless of the amount of sunlight and temperature. 'Breaking of dried stalks', which utilizes the physical power of 'threshing with a flail' is considered a core process of the fiber scutching technique in 'Yukjin' in Hamgyeong-do. The bark and the core of the hemp were separated by tapping, the bast was thinly split, and the shell was peeled off, making it suitable for collecting with thread. The method of collecting the fibers by applying physical power causes downing on the fibers, which is to be generally avoided in the manufacture of bast fabric woven hemp or ramie. However, Hamgyeong-do's fiber scutching method seems to have applied this principle to the method of making fragile fabrics by using it in reverse. This method is distinct from the steaming or boiling of the stalks' in Andong, Korea, and it is similar to the Western method of spinning fabrics.

Clinical Implication of Serum TNF-$\alpha$ and IL-1$\beta$ Measurement in Patients with Sepsis (패혈증환자에서 혈청 TNF-$\alpha$ 및 IL-1$\beta$)

  • Kim, Jae-Yeol;Choi, Hyung-Seok;Lee, Choon-Taek;Kim, Young-Whan;Han, Sung-Koo;Min, Kyung-Up;Kim, Yoo-Young;Shim, Young-Soo;Yoo, Chul-Gyu
    • Tuberculosis and Respiratory Diseases
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    • v.49 no.2
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    • pp.217-224
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    • 2000
  • Background : It is well known that when macrophages are stimulated with endotoxin, they produce a wide variety of cytokine mediators, including TNF-$\alpha$ and IL-1$\beta$. However, there is an alteration in the macrophages' responsiveness when they are challenged with repeated bouts of endotoxin, termed "endotoxin tolerance" which is regarded as a self-protective phenomenon from continuous stimulation. In this study, endotoxin tolerance in the peripheral blood monocytes of sepsis patients was evaluated. Methods : Fourteen patients with organism-documented sepsis were included. The severity of illness was evaluated by APACHE II score. Peripheral blood monocytes were isolated from the patients and diluted to $1{\times}10^5$ well. After stimulation with endotoxin (LPS of E. coli O114 : B4, 100 ng/ml), they were incubated at $37^{\circ}C$ in 5% $CO_2$ incubator for 24 hours. Supernatant was collected for the measurement of TNF-$\alpha$ and IL-1$\beta$ with ELISA method. Peripheral blood monocytes of seven healthy volunteers were used as control. Results : The APACHE II score (mean$\pm$SD) of the patients at the time of blood sampling was 12.2$\pm$5.7. The primary infection foci were urinary tract infection, pneumonia, subacute bacterial endocarditis, and catheter related infection, etc. The causative organisms were gram negative rods (10 cases), gram positive cocci (6 cases) with two cases of mixed infection. Serum TNF-$\alpha$ could be measured in 4 cases with 29.9$\pm$27.7 pg/ml. Serum IL-1$\beta$was measurable in only one patient. The TNF-$\alpha$ level of supernatant of cultured peripheral blood monocytes was 2,703$\pm$2,066 pg/ml in patients and 2,102$\pm$1914 pg/ml in controls. The IL-1$\beta$level of supernatant was 884$\pm$1,050 pg/ml in patients and 575$\pm$558 pg/ml in controls. There was no difference of TNF-$\alpha$ and IL-1$\beta$ level between patients and controls. Conclusion : We cannot prove the phenomenon of endotoxin tolerance in this study. Future study needs to be focused on the more severe sepsis patients who were taken for sampling earlier. Addition of serum to the culture medium could be an another valuable option for the success of this study.

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A Legal Study on Safety Management System (항공안전관리에 관한 법적 고찰)

  • So, Jae-Seon;Lee, Chang-Kyu
    • The Korean Journal of Air & Space Law and Policy
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    • v.29 no.1
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    • pp.3-32
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    • 2014
  • Safety Management System is the aviation industry policy for while operating the aircraft, to ensure the safety crew, aircraft and passengers. For operating a safe aircraft, in order to establish the international technical standards, the International Civil Aviation Organization has established the Annex 19 of the Convention on International Civil Aviation. As a result, member country was supposed to be in accordance with the policy of the International Civil Aviation Organization, to accept the international standard of domestic air law. The South Korean government announced that it would promote active safety management strategy in primary aviation policy master plan of 2012. And, by integrating and state safety programmes(ssp) and safety management system(sms) for the safe management of Annex 19 is to enforce the policy on aviation safety standards. State safety programmes(ssp) is a system of activities for the aim of strengthening the safety and integrated management of the activities of government. State safety programmes(ssp) is important on the basis of the data of the risk information. Collecting aviation hazard information is necessary for efficient operation of the state safety programmes(ssp) Korean government must implement the strategy required to comply with aviation methods and standards of the International Civil Aviation Organization. Airlines, must strive to safety features for safety culture construction and improvement of safety management is realized. It is necessary to make regulations on the basis of the aviation practice, for aviation safety regulatory requirements, aviation safety should reflect the opinion of the aviation industry.

Cytotoxic Effect of Taxol on Malignant Bone Tumor Cell Lines (악성 골종양 세포주들에 대한 Taxol의 세포독성)

  • Shin, Duk-Seop;Kim, Se-Dong;Kim, Keon-Ho;Lee, Jong-Hyung;Kim, Seong-Yong;Kim, Jung-Hye
    • The Journal of the Korean bone and joint tumor society
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    • v.4 no.1
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    • pp.13-21
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    • 1998
  • Taxol, the extract from the Taxus brevifolia which is a Pacific yew tree has aroused the interest of the tumor investigators since the 1960s. As well, it is shown to have broad antitumor activity in preclinical experimental models. Its action mechanism is an anti-microtubule effect by duplication of tubulin. The most impressive antitumor activity of taxol has been observed in advanced ovarian cancer and metastatic breast cancer. The purpose of this study was to determine how taxol acts on malignant bone tumor cell lines, to compare its cytotoxic effect with those of other chemotherapeutic agents, and to ascertain the its combination effect with adriamycin. Cell lines used in this study were G-292(osteosarcoma, human), SaOS-2(osteosarcoma, primary, human), and HT-1080(fibrosarcoma, human). Methotrexate, adriamycin, cisplatinum, ifosfamide and taxol were used as testing chemotherapeutic agents and their maximum test concentration were $500{\mu}g/ml$, $200{\mu}g/ml$, $500{\mu}g/ml$, $1000{\mu}g/ml$, and $600{\mu}g/ml$, respectively. The media for cell culture was RPMI-1640 with 10% fetal bovine serum and gentamycin. The results were as follows. The $IC_{50}$ of methotrexate, ifosfamide, cisplatinum, adriamycin and Taxol in G-292 were $2.3{\times}10^{-1}{\mu}g/ml$, $8.0{\times}10^0{\mu}g/ml$, $3.5{\times}10^0{\mu}g/ml$, $9.8{\times}10^{-1}{\mu}g/ml$, $2.7{\times}10^{-2}{\mu}g/ml$ respectively, in SaOS-2 $3.5{\times}10^{-1}{\mu}g/ml$, $1.5{\times}10^1{\mu}g/ml$, $2.8{\times}10^0{\mu}g/ml$, $9.9{\times}10^{-2}{\mu}g/ml$, $1.0{\times}10^{-2}{\mu}g/ml$, respectively, in HT-1080 $4.2{\times}10^{-2}{\mu}g/ml$, $5.4{\times}10^1{\mu}g/ml$, $3.8{\times}10^0{\mu}g/ml$, $5.5{\times}10^{-3}{\mu}g/ml$, $1.1{\times}10^{-3}{\mu}g/ml$, respectively. In conclusion, taxol had very potent cytotoxic effect on the malignant bone tumor cell lines with adriamycin, and was more potent than methotrexate, cisplatinum and ifosfamide. There were synergistic antitumor effects on G-292 and SaOS-2 cell lines in combination test of taxol and adriamycin. From the above results, it would be estimated that taxol could be a new antitumor drug for the malignant bone tumors, providing measures against the side effects and followed by the clinical tests.

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