• Title/Summary/Keyword: Development factor

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Activation of the NF-$\kappa$B p50/p65 Complex in Human Lung Cancer Cell Lines (인체 폐암세포주에서 NF-$\kappa$B p50/p65 Complex의 활성화)

  • Choi, Hyung-Seok;Yoo, Chul-Gyu;Lee, Choon-Taek;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo
    • Tuberculosis and Respiratory Diseases
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    • v.46 no.2
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    • pp.185-194
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    • 1999
  • Background: NF-$\kappa$B is a characteristic transcriptional factor whose functional activity is determined by post-translational modification of protein and subsequent change of subcellular localization. The involvement of the NF-$\kappa$B family of the transcription factors in the control of such vital cellular functions as immune response, acute phase reaction, replication of certain viruses and development and differentiation of cells has been clearly documented in many previous studies. Several recent observations have suggested that the NF-$\kappa$B might also be involved in the carcinogenesis of some hematological and solid tumors. Investigating the possibility that members of the NF-$\kappa$B family participate in the molecular control of malignant cell transformation could provide invaluable information on both molecular pathogenesis and cancer-related gene therapy. Method: To determine the expression patterns and functional roles of NF-$\kappa$B family transcription factors in human lung cancer cell lines NCI-H792, NCI-H709, NCI-H226 and NCI-H157 were analysed by western blot, using their respective antibodies. The nuclear and the cytoplasmic fraction of protein extract of these cell lines were subsequently obtained and NF-$\kappa$B expression in each fraction was again determined by western blot analysis. The type of NF-$\kappa$B complex present in the cells was determined by immunoprecipitation. To detect the binding ability of cell-line nuclear extracts to the KB consensus oligonucleotide, electrophoretic mobility shift assay(EMSA) was performed. Results: In the cultured human lung cancer cell lines tested, transcription factors of the NF-$\kappa$B family, namely the p50 and p65 subunit were expressed and localized in the nuclear fraction of the cellular extract by western blot analysis and immunocytochemistry. Immunoprecipitation assay showed that in the cell, the p50 and p65 subunits made NF-$\kappa$B complex. Finally it was shown by Electrophoretic Mobility Shift Assay(EMSA) that nuclear extracts of lung cancer cell lines are able to bind to NF-$\kappa$B consensus DNA sequences. Conclusion: These data suggest that in human lung cancer cell lines the NF-$\kappa$B p50/p65 complex might be activated. and strengthen the hypothesis that NF-$\kappa$B family transcription factors might be involved in the carcinogenesis of human lung cancer.

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Development of Movement Analysis Program and its Feasibility Test in Streotactic Body Radiation Threrapy (복부부위의 체부정위방사선치료시 호흡에 의한 움직임분석 프로그램 개발 및 유용성 평가)

  • Shin, Eun-Hyuk;Han, Young-Yih;Kim, Jin-Sung;Park, Hee-Chul;Shin, Jung-Suk;Ju, Sang-Gyu;Lee, Ji-Hea;Ahn, Jong-Ho;Lee, Jai-Ki;Choi, Doo-Ho
    • Progress in Medical Physics
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    • v.22 no.3
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    • pp.107-116
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    • 2011
  • Respiratory gated radiation therapy and stereotactic body radiation therapy require identical tumor motions during each treatment with the motion detected in treatment planning CT. Therefore, this study developed a tumor motion monitoring and analysis system during the treatments employing RPM data, gated setup OBI images and a data analysis software. A respiratory training and guiding program which improves the regularity of breathing was used to patients. The breathing signal was obtained by RPM and the recorded data in the 4D console was read after treatment. The setup OBI images obtained gated at 0% and 50% of breathing phases were used to detect the tumor motion range in crenio-caudal direction. By matching the RPM data recorded at the OBI imaging time, a factor which converts the RPM motion to the tumor motion was computed. RPM data was entered to the institute developed data analysis software and the maximum, minimum, average of the breathing motion as well as the standard deviation of motion amplitude and period was computed. The computed result is exported in an excel file. The conversion factor was applied to the analyzed data to estimate the tumor motion. The accuracy of the developed method was tested by using a moving phantom, and the efficacy was evaluated for 10 stereotactic body radiation therapy patients. For the sine wave motion of the phantom with 4 sec of period and 2 cm of peak-to-peak amplitude, the measurement was slightly larger (4.052 sec) and the amplitude was smaller (1.952 cm). For patient treatment, one patient was evaluated not to qualified to SBRT due to the usability of the breathing, and in one patient case, the treatment was changed to respiratory gated treatment due the larger motion range of the tumor than treatment planed motion. The developed method and data analysis program was useful to estimate the tumor motion during treatment.

Growth Effect of Oncorhychus masou by Recombinant Myostatin Prodomain Proteins Derived from Fish (어류 유래 마이오스타틴 프로도메인 단백질에 의한 시마연어(Oncorhychus masou) 성장효과)

  • Kim, Jeong-Hwan;Lee, Sang-Beum;Cho, Mi-Jin;Ahn, Ji-Young;Lee, Suk-Keun;Hong, Sung-Youl;Seong, Ki-Baik;Jin, Hyung-Joo
    • Journal of Life Science
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    • v.21 no.8
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    • pp.1149-1155
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    • 2011
  • Myostatin (MSTN) belongs to the transforming growth factor-${\beta}$ superfamily or growth and differentiation factor 8 (GDF-8), and functions as a negative regulator of skeletal muscle development and growth. Previous studies in mammals have suggested that myostatin knock-out increased muscle mass and decreased fat content compared to those of the wide type. Recently, several studies on myostatin have beenconducted on the block myostatin signal pathway with myostatin antagonists and the MSTN regulation with RNAi to control myostatin function. This study was performed to analyze growth and muscle alteration of Oncorhychus masou by treatment with recombinant myostatin prodomains derived from fish. We designed myostatin prodomains derived from P. olivaceus (pMALc2x-poMSTNpro) and S. schlegeli (pMALc2x-sMSTNpro) in a pMALc2x expression vector, and then purified the recombinant proteins using affinity chromatography. The purified recombinant proteins were treated in O. masou through an immersion method. Recombinant protein treated groups did not show a significant difference in weight, protein, or lipid composition compared to the control. However, there was a difference in the average number and area for histological analyses in the muscle fiber. At twelve and twenty-two weeks from the initial treatment, there were differences in averagefiber number and area between the 0.05 mg/l treated-group and the control, but the numbers were similar to those of the control during the same time period. At twelve weeks, however, 0.2 mg/l treated-group had an increase in average fiber number and decrease in average fiber area compared to the control. At twenty-two weeks, the pMALc2x-sMSTNpro 0.2 mg/l treated-group was induced and showed a decrease in average fiber number and increase in average fiber area. The results between twelve and twenty-two weeks showed that the fiber numbers had decreased, whereas average fiberarea had increased due to sMSTNpro. It is understood that the sMSTNpro induced only hyperplasia at twelve weeks, after which it induced hypertrophy. Recombinant myostatin prodomains derived from fish may induce hyperplasia and hypertrophy in O. masou depending upon the time that has elapsed.

Effect of Steviol β-Glucopyranosyl Ester on The Production of Nitric Oxide and Inflammatory Cytokines in RAW 264.7 Cells (Steviol β-Glucopyranosyl Ester가 RAW 264.7 세포의 산화질소 및 염증성 사이토카인 생성에 미치는 영향)

  • Jung, Heehoon;Cho, Uk Min;Hwang, Hyung Seo;Cho, Kun;Lee, Sang Rin;Kim, Moo Sung
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.44 no.3
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    • pp.239-247
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    • 2018
  • Chronic inflammation is known to have effects on various diseases such as gout, cancer, dementia, atopic disease, and obesity. In addition, since some signal cascades involved in the development of inflammation are known to affect the damage and aging of the skin tissue, studies are being conducted actively to control the inflammation mechanism. In order to mitigate or prevent inflammatory response, a number of researches have been made to develop anti-inflammatory materials from some plants. In particular, Stevia rebaudiana produces steviol glycosides (SG), a natural sweetener with a distinctive flavor. Studies on some of SG have been shown to have anti-inflammatory activity. Researchers of this study expected that more SG also possess anti-inflammatory activity, besides stevioside, rebaudioside A, and steviol. In order to confirm this possibility, the researchers screened inhibition activity of various steviol glucosides for NO production in RAW 264.7 cell lines. As a result, steviol ${\beta}-glucopyranosyl$ ester (SGE) showed the highest inhibitory activity among steviol derivatives treated at the same molar concentration. In addition, we found that mRNA expression level of $interleukin-1{\alpha}$ ($IL-1{\alpha}$), $interleukin-1{\beta}$ ($IL-1{\beta}$), cyclooxygenase-2 (COX-2), nuclear factor kappa-light chain-enhancer of activated B cells ($NF-{\kappa}B$) and inducible nitric oxide synthase (iNOS) was also decreased in a dose-dependent manner. These results show that SGE inhibits anti-inflammatory activity and NO production in mouse macrophage RAW 264.7 cells. It was confirmed that SGE has potential to be applied as an anti-inflammatory material.

Expression of Osteopontin and Transforming Growth Factor- ${\beta}$ in Childhood Minimal Change Nephrotic Syndrome After Cyclosporine Treatment (미세변화 신증후군 환아에서 사이클로스포린 치료 후 Osteopontin과 Transforming Growth Factor-${\beta}$의 발현)

  • Lim Beom-Jin;Kim Pyung-Kil;Hong Soon-Won;Jeong Hyeon-Joo
    • Childhood Kidney Diseases
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    • v.6 no.2
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    • pp.142-154
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    • 2002
  • Purpose : One of the most important adverse effects of long-term cyclosporine therapy is nephrotoxicity, the morphologic changes of which include interstitial fibrosis and arteriolar hyalinization. Recently, several authors have shown that osteopontin plays an important role in the development of interstitial fibrosis by acting as a macrophage chemoattractant and stimulating the production of $TGF-{\beta}$ in experimental cyclosporine nephrotoxicity. However, the relationship between osteopontin and $TGF-{\beta}$ in humans has not been clearly documented so far. We studied the expression of osteopontin and $TGF-{\beta}$ in children with minimal change nephrotic syndrome treated with cyclosporine to demonstrate whether there is a relationship between cyclosporine toxicity and osteopontin expression as previously shown in animal models. Materials and methods : Nineteen children (15 males and 4 females) were the subject of this study. Renal biopsies had been performed before and after the cyclosporine therapy (mean duration: 15.9 months). In 5 patients, additional biopsies were performed after completing the cyclosporine treatment (mean; 26 months). The expressions of osteopontin and $TGF-{\beta}$ were evaluated by immunohistochemistry in the glomeruli and tubulointerstitium. Results : Osteopontin expression was significantly increased in the glomerular mesangium and tubules after cyclosporine treatment. But there was no statistically significant increase of $TGF-{\beta}$ in the interstitium. There was no significant increase in tubular osteopontin and interstitial $TGF-{\beta}$ expression in those cases developing interstitial fibrosis after cyclosporine treatment compared with cases those not developing interstitial fibrosis. No significant changes in osteopontin or $TGF-{\beta}$ expression were observed in subsequent 5 biopsy samples after discontinuation of cyclosporine compared with the first follow up biopsies. Conclusion : These results suggest that osteopontin is a nonspecific marker of renal injury rather than a mediator of interstitial fibrosis in cyclosporine nephrotoxicity of human.

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The Usefulness of Visceral Fat Thickness Measured by Ultrasonography as an Abdominal Obesity Index (초음파로 측정된 내장지방두께의 복부 비만지표로서의 유용성)

  • Kim, Yong-Kyun;Han, Man-Seok
    • Journal of radiological science and technology
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    • v.31 no.3
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    • pp.249-258
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    • 2008
  • Purpose : Abdominal obesity with visceral fat accumulation have been known to be intimately associated with the development of metabolic syndrome. Therefore, it is important to estimate the precise amount of visceral fat. Ultrasonography has been reported that it is a simple and noninvasive method for visceral fat evaluation. Purpose of this study is to evaluate the association of ultrasonographic visceral fat thickness, anthropometric indexes, and risk factor of metabolic syndrome, and to investigate the cut-off value of abdominal visceral fat thickness leading to increased risk of metabolic syndrome. Materials and methods : The subject included 200 men and 200 women who visited D healthcare center in Daejeon from January to April 2008. The subcutaneous fat thickness and visceral fat thickness were measured by ultrasonograph. As anthropometric index, we measured body mass index, waist circumference and waist/height ratio. As for the risk factor of metabolic syndrome, we measured blood pressure, high density lipoprotein cholesterol, triglyceride and fasting serum glucose. Results : VFT was significantly correlated with waist circumference, (r=0.683/M, r=0.604/F), waist to height ratio (r=0.633/M, r=0.593/F) and BMI (r=0.621/M, r=0.534/F) in both men and women. In addition it was significantly correlated with Systolic blood pressure (r=0.229/M, r=0.232/F), Diastolic blood pressure ((r=0.285/M, r=0.254/F), high density cholesterol (r=-0.254/M, r=-0.254/F), Triglyceride (r=0.475/M, r=0.411/F), and Fasting blood sugar (r=0.158/M, r=0.234/F) in both men and women. The cut-off value of visceral fat thickness leading to the increased risk of metabolic syndrome was 4.58cm (sensitivity89.2%, specificity 71.2%) in men and 3.50cm (sensitivity61.2% specificity 80.8%) in women respectively. The odds ratio of the risk of metabolic syndrome was dramatically increased with the abdominal visceral fat thickness level over 6cm in men and 5cm in women. Conclusion : The visceral fat thickness using ultrasonography was significantly correlated with anthropometric indexes and risk factors of metabolic syndrome in both men and women. The cut-off value of visceral fat thickness leading to the increased risk of metabolic syndrome was 4.58cm in men and 3.50cm in women.

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Hypothyroidism Following Surgery and Radiation Therapy for Head and Neck Cancer (두경부암 환자에서 수술 및 방사선치료 후 갑상선 기능 저하)

  • Park, In-Kyu;Kim, Jae-Cheol
    • Radiation Oncology Journal
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    • v.15 no.3
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    • pp.225-231
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    • 1997
  • Purpose : Radiation therapy in combination with surgery has an important role in the therapy of the head and neck cancer We conducted a prospective study for patients with head and neck cancer treated with surgery and radiation to evaluate the effect of therapies on the thyroid gland, and to identify the factors that might influence the development of hypothyroidism. Materials and Methods : From September 1986 through December 1994, 71 patients with head and cancer treated with surgery and radiation were included in this prospective study. Patients' age ranged from 32 to 73 years with a median age of 58 years. There were 12 women and 59 men. The primary tumor sites were larynx in 34 patients, hypopharynx in 13 patients, oral cavity in 12 patients, unknown primary of the neck in 6 patients, salivary gland in 3 patients, maxillary sinus in 2 patients, and oropharynx in 1 patient. Total laryngectomy with neck dissection was carried out in 45 patients and neck dissection alone in 26 patients. All patients were serially monitored for thyroid function (T3, T4, free T4, TSH, antithyroglobulin antibody and antimicrosomal antibody) before and after radiation therapy. Radiation dose to the thyroid gland ranged from 40.6Gy to 60Gy with a median dose of 50Gy The follow-up duration was 3 to 80 months. Results :The overall incidence of hypothyroidism was 56.3\%$);7 out of 71 patients $(9.9\%)$ developed clinical hypothyroidism and 33 patients $(46.4\%)$ developed subclinical hypothyroidism. No thyroid nodules, thyroid cancers, or hyperthyroidism was detected. Hypothyroidism developed earlier in patients who underwent total laryngectomy with neck dissection than in patients with neck dissection alone (P<0.05). The risk factor that significantly influenced the incidence of hypothyroidism was a combination of surgery (total laryngectomy with neck dissection) and radiation therapy (P=0.0000), Four of 26 patients $(15.4\%)$ with neck dissection alone developed hypothyroidism while 36 of 45 patients $(80\%)$ with laryngectomy and neck dissection developed hypothyroidism. Conclusion : The hypothyroidism following surgery and radiation therapy was a relatively common complication. The factor that significantly influenced theincidence of hypothyroidism was combination of surgery and radiation therapy. Evaluation of thyroid function before and after radiation therapy with periodic thyroid function tests is recommended for an early detection of hypothyroidism and thyroid hormone replacement therapy is recommended whenever hypothyroidism develops.

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Nutritional and Tissue Specificity of IGF-I and IGFBP-2 Gene Expression in Growing Chickens - A Review -

  • Kita, K.;Nagao, K.;Okumura, J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.5
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    • pp.747-754
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    • 2005
  • Nutritional regulation of gene expression associated with growth and feeding behavior in avian species can become an important technique to improve poultry production according to the supply of nutrients in the diet. Insulin-like growth factor-I (IGF-I) found in chickens has been characterized to be a 70 amino acid polypeptide and plays an important role in growth and metabolism. Although it is been well known that IGF-I is highly associated with embryonic development and post-hatching growth, changes in the distribution of IGF-I gene expression throughout early- to late-embryogenesis have not been studied so far. We revealed that the developmental pattern of IGF-I gene expression during embryogenesis differed among various tissues. No bands of IGF-I mRNA were detected in embryonic liver at 7 days of incubation, and thereafter the amount of hepatic IGF-I mRNA was increased from 14 to 20 days of incubation. In eyes, a peak in IGF-I mRNA levels occurred at mid-embryogenesis, but by contrast, IGF-I mRNA was barely detectable in the heart throughout all incubation periods. In the muscle, no significant difference in IGF-I gene expression was observed during different stages of embryogenesis. After hatching, hepatic IGF-I gene expression as well as plasma IGF-I concentration increases rapidly with age, reaches a peak before sexual maturity, and then declines. The IGF-I gene expression is very sensitive to changes in nutritional conditions. Food-restriction and fasting decreased hepatic IGF-I gene expression and refeeding restored IGF-I gene expression to the level of fed chickens. Dietary protein is also a very strong factor in changing hepatic IGF-I gene expression. Refeeding with dietary protein alone successfully restored hepatic IGF-I gene expression of fasted chickens to the level of fed controls. In most circumstances, IGF-I makes a complex with specific high-affinity IGF-binding proteins (IGFBPs). So far, four different IGFBPs have been identified in avian species and the major IGFBP in chicken plasma has been reported to be IGFBP-2. We studied the relationship between nutritional status and IGFBP-2 gene expression in various tissues of young chickens. In the liver of fed chickens, almost no IGFBP-2 mRNA was detected. However, fasting markedly increased hepatic IGFBP-2 gene expression, and the level was reduced after refeeding. In the gizzard of well-fed young chickens, IGFBP-2 gene expression was detected and fasting significantly elevated gizzard IGFBP-2 mRNA levels to about double that of fed controls. After refeeding, gizzard IGFBP-2 gene expression decreased similar to hepatic IGFBP-2 gene expression. In the brain, IGFBP-2 mRNA was observed in fed chickens and had significantly decreased by fasting. In the kidney, IGFBP-2 gene expression was observed but not influenced by fasting and refeeding. Recently, we have demonstrated in vivo that gizzard and hepatic IGFBP-2 gene expression in fasted chickens was rapidly reduced by intravenous administration of insulin, as indicated that in young chickens the reduction in gizzard and hepatic IGFBP-2 gene expression in vivo stimulated by malnutrition may be, in part, regulated by means of the increase in plasma insulin concentration via an insulin-response element. The influence of dietary protein source (isolated soybean protein vs. casein) and the supplementation of essential amino acids on gizzard IGFBP-2 gene expression was examined. In both soybean protein and casein diet groups, the deficiency of essential amino acids stimulated chickens to increase gizzard IGFBP-2 gene expression. Although amino acid supplementation of a soybean protein diet significantly decreased gizzard IGFBP-2 mRNA levels, a similar reduction was not observed in chickens fed a casein diet supplemented with amino acids. This overview of nutritional regulation of IGF-I and IGFBP-2 gene expression in young chickens would serve for the establishment of the supply of nutrients to diets to improve poultry production.

The Study on the Influence of Selection Characteristics of Franchise System, business possibility, Communication, Moral Hazard on Franchisee's Perceived Risk, and Recontracting Intention in the Food Service Franchise Industry (외식 프랜차이저의 사업성, 커뮤니케이션, 모럴해저드가 프랜차이지의 위험지각과 재계약의도에 미치는 영향)

  • Yu, Jong-Pil;Lee, In-Ho
    • Journal of Distribution Research
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    • v.16 no.1
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    • pp.1-27
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    • 2011
  • I. Introduction: This study is to examine the structural relationships among exogenous variable (preliminary and post-support, franchisee's perceived business possibility, communication, moral hazard), the mediated variables(satisfaction, perceived risk, trust) and dependent variable(recontracting intention) in the food service franchise industry context. More specifically, this study has considered some realistic characteristics factors influencing satisfaction, perceived risk and trust between franchisors and franchisees and their further recontracting intention from the perspective of a practical approach. In this study, 437 data has been collected and used for the SPSS and AMOS analysis. The data were analyzed with structural equation modeling. Since the result of the overall model analysis demonstrated a good fit, we could further analyze our data. II. Research Model: This study is to examine the structural relationships among preliminary and post-support by franchisor, franchisee's perceived business possibility, and communication, moral hazard, has on effect on franchisee's satisfaction, perceived risk, trust and recontracting intention in the food service franchise industry context. Hypotheses are as following (Stern & EL-Ansary 1988; Oliver, 1997;Kee & Knox, 1970; Moorman, Deshpande & Zaltman, 1993; Perron, 1998; Zaheer, McEvily, Perrone, 1998). III. Result and Implication: We examined franchisee who have food service stores for samples of this study. The data were analyzed with structural equation modeling using path analysis. The result of the overall model analysis appeared as following: ${\chi}^2$ = 61.578 (d.f.=9, p<0.01), CFI =.990, GFI =.973, AGFI =.863, RMR =.019, RMSEA= .116, NFI = .988, TLI = .959. The findings can be summarized as follows: First, preliminary and post support of franchisor, perceived business possibility and communication positively influence to franchisee's satisfaction. Second, moral hazard of franchisor has negatively influence to franchisee's satisfaction and positively influence to perceived risk. Third, franchisee's satisfaction and trust has positively influence to recontracting intention. Fourth, franchisee's perceived risk has negatively influence to trust and recontracting intention. We can concluded that franchisor's preliminary and post support of franchisor, perceived business possibility and communication may be considered as the important factors influence to franchisee's satisfaction. Moral hazard has become a focused issue in franchise industry. Finally, the managerial implication has been stated as followings: First, in the process of building a systematic industry support franchise system and developing a creative business model, franchisee's stable profitability should be considered as the first important factor. The franchisee's trust to franchise may become a dominant factor that influence the business expansion of franchisor. Second, franchisor should communication with their franchisees and deal with the realistic difficulties faced by them with an effort. Third, the franchisor should achieve a synergy effect by utilizing the win-win strategy. The moral hazard strategy that achieving the profit through franchisee's damage will not be inadvisable to franchisor. Then the long-term oriented development and profitability can be maintained. To do so, the franchise industry may break away from the traditional business structure to improve management transparency and competitiveness on investment and organizational changing management. The conflict between franchisor and franchisee also can be reduced and big success can be achieved in the franchise industry.

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Ammonia Gas Removal by Bacillus subtilis IB101 and Dctimization of Culture Media (Bacillus subtilis IB101을 이용한 암모니아 가스 제거 및 생산배지 최적화)

  • Kim, So-Young;Noh, Yong-Ho;Kang, Sung-Gak;Kim, Young-Bum;Jang, Woo-Jin;Kim, Dong-Joon;Yun, Hyun-Shik
    • KSBB Journal
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    • v.22 no.3
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    • pp.162-167
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    • 2007
  • Ammonia gas is one of the major pollutants which cause environmental pollution and damage to the human and the livestock. The objective of this study was to investigate the important parameters for the development of efficient removal of ammonia gas by Bacillius subtilis IB101 and to optimize the medium composition for the mass production of B. subtilis IB101. The ammonia gas removal efficiency was evaluated at different growth phases and by changing culture conditions (temperature, pH). The effect of $(NH_4)_2SO_4$ concentration in preculture medium was examined. Medium optimization for the mass production of B. subtilis IB101 was performed by using Plackett-Burman design and one factor at a time method. The removal of ammonia gas was more efficient at exponential phase by 20% than at stationary phase. The ammonia gas removal was the highest at pH 4 and 30 $^{\circ}C$. There was not any significant influence of concentration of $(NH_4)_2SO_4$ on the removal of ammonia gas. The components of optimized medium for the production of viable Bacillus subtilis IB101 was yeast extract 10 g/l, soluble starch 2.5 g/l, $MgSO_4$ 6 g/l, $CaCl_2$ 1.55 g/l, $(NH_4)_2SO_4$ 5 g/l, $KH_2PO_4$ 0.75 g/l, soy bean meal 8 g/l.