• Title/Summary/Keyword: cellular level

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Physiological Changes of Saccharomyces cerevisiae KNU5377 Occurred in the Process of the 48-hour Ethanol Fermentation at 40℃ (40℃ 48시간 에탄올발효 과정 중 일어나는 Saccharomyces cerevisiae KNU5377의 생리 변화)

  • Kwak, Sun-Hye;Kim, Il-Sup;Kang, Kyung-Hee;Lee, Jung-Sook;Jin, Ingn-Yol
    • Journal of Life Science
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    • v.21 no.1
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    • pp.146-154
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    • 2011
  • In this study, physiological changes in a thermotolerant yeast Saccharomyces cerevisiae KNU5377 cell exposed to 48-hour alcohol fermentation at $40^{\circ}C$ were investigated. After 12 hours of alcohol fermentation at $40^{\circ}C$, the $C_{16:1}$ unsaturated acid of plasma membrane increased to 1.5 times more than the $C_{16:0}$ saturated fatty acid, and to about 2 times more for the $C_{18:1}$ unsaturated fatty acid. Fermentation at both $30^{\circ}C$ and $37^{\circ}C$ fermentation showed the same pattern as that done at $40^{\circ}C$. The pH of the alcohol-fermentation medium was reduced to pH 4.1 from a starting pH of 6.0 through the 12-hr fermentation and then maintained this level during the continuing fermentation. With the process of fermentation, the remaining glucose was reduced, but its amount remaining during the $40^{\circ}C$-fermentation was less reduced than those fermented at $30^{\circ}C$ and $37^{\circ}C$. In the study investigating the changing pattern of cellular proteins in the alcohol-fermenting cells, the SDS-PAGE and 2-D data indicated the most expressed dot was phosphoglycerate kinase, which is one enzyme involved in glycolysis. Why this enzyme was most expressed in the cells exposed to unfavorable conditions such as high temperature, increasing concentration of produced alcohol and long time exposure to other stress factors remains unsolved.

Expressional Analysis of Two Genes (Got1 andMat1) Up-regulated by Starvation Stress (영양고갈-스트레스에 의해서 상승 발현하는 유전자(Got1과 Mat1)의 분석)

  • Park, Junseok;Kwon, Young-Sook;Lee, Eunryoung;Kwon, Kisang
    • Journal of Life Science
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    • v.24 no.6
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    • pp.686-693
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    • 2014
  • Restricted supply of nutrients may affect genes at the molecular level as well as physiological functions. Understanding the cellular responses during starvation is necessary for developing strategies to reduce damage caused by starvation stress. After 1 h of starvation, Got1 gene expression was increased but its expression returned to the normal state after 24 h. Mat1 gene expression continuously increased with starvation from 1 h until 24 hr. Rats starved for 1-3 days showed significant changes in expression of the Got1 and Mat1 genes, which were significantly reduced in the cerebral cortex and cerebellum. In the lung, gene expression was increased by starvation for 1-2 days but decreased on the third day. No differences were observed in gene expression in the heart. Strong Got1 lung gene expression was seen in the starvation group one day after restoration of the food supply. Muscle mass was significantly reduced at the start of starvation and remained the same after two days of starvation and one day after the food supply was restored. The Mat1 gene expression did not change. The Got1 was induced by NaCl and showed strong expression in the lung and the thymus, but the apparent decrease of the remaining changes were not observed in male rats. The Mat1 gene was not as sensitive as the Got1 gene to induction by NaCl. However, differences in gene induction by NaCl were evident between males and females, indicating that diet control of gene expression is associated with hormones.

Effects of Recombinant Human Epidermal Growth Factor (rhEGF) on Experimental Radiation-Induced Oral Mucositis in Rats (Rat의 방사선 조사성 구내염에 대한 Recombinant Human Epidermal Growth Factor (rhEGF)의 효과)

  • Jung Kwon-Il;Kim Sun-Hee;Moon Soo-Young;Kim Yeon-Wha;Hong Joon-Pio;Kim Hyun-Sook;Lee Sang-Wook
    • Radiation Oncology Journal
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    • v.24 no.1
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    • pp.67-76
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    • 2006
  • Purpose: Oral mucositis is a common toxicity of radiation or chemotherapy, which is used a treatment for head and neck cancer. We investigated effects of recombinant human epidermal growth factor (rhEGF) on radiation-induced oral mucositis in rat model. Materials and Methods: Spraque-Dawley rats (7 per group) exposed to a single dose of 25 Gy (day 0) on their head, except for one group, were randomly divided into un-treated, vehicle-treated, and two rhEGF-treated groups. Rats were topically applied with rhEGF (15 or $30{\mu}g/oral$ cavity/day) or vehicle to their oral mucosa. Survival rate of rats, weight changes, and food intakes were examined from day 0 to 18 after radiation. Histology study was performed from oral mucosa of rats at day 7 and 18 after radiation. Results: rhEGF-treated groups (15 or $30{\mu}g/oral$) showed all survival rate 33%, whereas un-treated and vehicle-treated groups showed all survival rate 0% at the end of experiment. rhEGF-treated groups statistically had less weight loss compared to vehicle-treated group from day 2 to 7 after radiation. Food intake of rats with rhEGF treatment turned to increase at day 14 after radiation. At 7 day after radiation, un-treated and vehicle-treated groups showed severe pseudomembraneous or ulcerative oral mucositis. On the other hand, rhEGF-treated groups had no more than cellular swelling and degeneration of epidermal cells in oral mucosa of rats. Conclusion: These results suggest that rhEGF has significantly positive effects on radiation-induced oral mucositis in rats. rhEGF display a therapeutic potential on a clinical level.

A Clinical and Pathological Analysis of Children with Membranoproliferative Glomerulonephritis According to the Clinical Manifestations at Presentation (발견 양상에 따른 소아 막증식성 사구체신염의 임상적 및 병리조직학적분석)

  • Jeon Chang-Ho;Kang Mi-Seon;Chung Woo-Yeong
    • Childhood Kidney Diseases
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    • v.8 no.2
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    • pp.186-194
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    • 2004
  • Purpose: Membranoproliferative glomeulonephritis(MPGN) has been diagnosed in an increasing number of asymptomatic cases. These cases have been detected by school urinary screening test even though the total cases of MPGN show a decreasing trend. We have analyzed the clinical and pathological characteristics of children with MPGN according to the clinical manifestations at the time of disease presentation. Methods: A total of 18 patients who had been diagnosed with idiopathic MPGN by percutaneous renal biopsy from January 1990 to February 2004 were involved in our study. The patients were divided into 2 groups as the school urinary screening(A) group and the symptomatic(S) group according to the clinical manifestations at the time of disease presentation. Results: Out of the total 18 patients, 8(44.4%) were in the S group and 10(55.6%) were in the A group. The mean serum total protein, albumin and $C_3$ levels in the S group were significantly lower than those levels of the A group, respectively($4.9{\pm}1.2\;g/dL,\;vs\;7.0{\pm}0.5\;g/dL\;P=0.002,\;2.8{\pm}0.9\;g/dL\;vs.\;4.1{\pm}0.3\;g/dL\;P=0.002,\;63.9{\pm}36.4\;mg/dL\;vs.\;100.8{\pm}39.5\;g/dL\;P=0.041$). The mean total protein amount of 24 hour collected urine in the S group were significantly higher than that of the A group($3684.0{\pm}2601.3\;mg/m^2\;vs.\;559.4{\pm}4.6.9\;mg/m^2$, respectively, P=0.001). Hypocomplementemia was observed in 11(61.1%) out of 18 patients at the time of disease onset, 7(87.5%) in the S group and 4(40%) in the A group. However the hypocomplementemia was decreased in 6(33.3%) out of 18 patients at the time of final follow-up, 3(37.5%) in the S group and 3(30%) in the A group. According to the pathologic type, hypocomplementemia was observed 8 patients(61.5%) with type I disease, 1 patients (100%) with type II disease, 2 patients(50%) in type III disease at the disease onset, but 4 patients(30.8%) in type I disease, 1 patient(100%) in type II disease, 1 patient(33.3%) with type III disease at the time of last follow-up. The incidence of cellular crescent formation and tubular atropy. as observed on light microscopy, were higher in the S group compared to the A group. Mean grade of capillary wall thickening and, mesangial proliferation were significantly higher in the S group. Conclusion: MPGN, as diagnosed in patients with only asymptomatic urinary abnormalities, has been increasing, it is more frequent in asymptomatic patients than in patients with presenting symptoms. Our result suggests that MPGN should be considered in the renal biopsy diagnosis regardless of serum $C_3$ level when urinary abnormalities are found by school urinary screening test.

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Gene Expression Profiles in Cervical Cancer with Radiation Therapy Alone and Chemo-radiation Therapy (자궁경부암의 방사선치료 및 방사선항암화학 병용치료에 따른 유전자발현 조절양상)

  • Lee Kyu Chan;Kim Meyoung-kon;Kim Jooyoung;Hwang You Jin;Choi Myung Sun;Kim Chul Yong
    • Radiation Oncology Journal
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    • v.21 no.1
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    • pp.54-65
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    • 2003
  • Purpose : To analyze the gene expression Profiles of uterine ceulcal cancer, and its variation after radiation therapy, with or without concurrent chemotherapy, using a CDNA microarray. Materials and Methods :Sixteen patients, 8 with squamous ceil carcinomas of the uterine cervix, who were treated with radiation alone, and the other 8 treated w14h concurrent chemo-radiation, were Included in the study. Before the starling of the treatment, tumor biopsies were carried out, and the second time biopsies were peformed after a radiation dose of 16.2$\~$27 Gy. Three normal cervix tissues were used as a control group. The microarray experiments were peformed with 5 groups of the total RNAs extracted individually and then admixed as control, pre-radiation therapy alone, during-radiation therapy alone, pre-chemoradiation therapy, and during-chemoradlation therapy. The 33P-iabeled CDNAS were synthesized from the total RNAs of each group, by reverse transcription, and then they were hybridized to the CDNA microarray membrane. The gene expression of each microarrays was captured by the intensity of each spot produced by the radioactive isotopes. The pixels per spot were counted with an Arrayguage, and were exported to Microsoft Excel The data were normalized by the Z transformation, and the comparisons were peformed on the Z-ratio values calculated. Results : The expressions of 15 genes, including integrin linked kinase (ILK), CDC28 protein kinase 2, Spry 2, and ERK 3, were increased with the Z-ratio values of over 2.0 for the cervix cancer tissues compared to those for the normal controls. Those genes were involved In cell growth and proliferation, cell cycle control, or signal transduction. The expressions of the other 6 genes, Including G protein coupled receptor kinase 5, were decreased with the Z-ratio values of below -2.0. After the radiation thorapy, most of the genes, with a previously Increase expressions, represented the decreased expression profiles, and the genes, with the Z-ratio values of over 2.0, were cyclic nucleotlde gated channel and 3 Expressed sequence tags (EST). In the concurrent chemo-radiation group, the genes involved in cell growth and proliferation, cell cycle control, and signal transduction were shown to have increased expressions compared to the radiation therapy alone group. The expressions of genes involved in anglogenesis (angiopoietln-2), immune reactions (formyl peptide receptor-iike 1), and DNA repair (CAMP phosphodiesterase) were increased, however, the expression of gene involved In apoptosls (death associated protein kinase) was decreased. Conclusion : The different kinds of genes involved in the development and progression of cervical cancer were identified with the CDNA microarray, and the proposed theory is that the proliferation signal stalls with ILK, and is amplified with Spry 2 and MAPK signaling, and the cellular mitoses are Increased with the increased expression oi Cdc 2 and cell division kinases. After the radiation therapy, the expression profiles demonstrated 4he evidence of the decreased cancer cell proliferation. There was no sigificant difference in the morphological findings of cell death between the radiation therapy aione and the chemo-radiation groups In the second time biopsy specimen, however, the gene expression profiles were markedly different, and the mechanism at the molecular level needs further study.

The Ability of Anti-tumor Necrosis Factor Alpha(TNF-${\alpha}$) Antibodies Produced in Sheep Colostrums

  • Yun, Sung-Seob
    • 한국유가공학회:학술대회논문집
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    • 2007.09a
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    • pp.49-58
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    • 2007
  • Inflammatory process leads to the well-known mucosal damage and therefore a further disturbance of the epithelial barrier function, resulting abnormal intestinal wall function, even further accelerating the inflammatory process[1]. Despite of the records, etiology and pathogenesis of IBD remain rather unclear. There are many studies over the past couple of years have led to great advanced in understanding the inflammatory bowel disease(IBD) and their underlying pathophysiologic mechanisms. From the current understanding, it is likely that chronic inflammation in IBD is due to aggressive cellular immune responses including increased serum concentrations of different cytokines. Therefore, targeted molecules can be specifically eliminated in their expression directly on the transcriptional level. Interesting therapeutic trials are expected against adhesion molecules and pro-inflammatory cytokines such as TNF-${\alpha}$. The future development of immune therapies in IBD therefore holds great promises for better treatment modalities of IBD but will also open important new insights into a further understanding of inflammation pathophysiology. Treatment of cytokine inhibitors such as Immunex(Enbrel) and J&J/Centocor(Remicade) which are mouse-derived monoclonal antibodies have been shown in several studies to modulate the symptoms of patients, however, theses TNF inhibitors also have an adverse effect immune-related problems and also are costly and must be administered by injection. Because of the eventual development of unwanted side effects, these two products are used in only a select patient population. The present study was performed to elucidate the ability of TNF-${\alpha}$ antibodies produced in sheep colostrums to neutralize TNF-${\alpha}$ action in a cell-based bioassay and in a small animal model of intestinal inflammation. In vitro study, inhibitory effect of anti-TNF-${\alpha}$ antibody from the sheep was determined by cell bioassay. The antibody from the sheep at 1 in 10,000 dilution was able to completely inhibit TNF-${\alpha}$ activity in the cell bioassay. The antibodies from the same sheep, but different milkings, exhibited some variability in inhibition of TNF-${\alpha}$ activity, but were all greater than the control sample. In vivo study, the degree of inflammation was severe to experiment, despite of the initial pilot trial, main trial 1 was unable to figure out of any effect of antibody to reduce the impact of PAF and LPS. Main rat trial 2 resulted no significant symptoms like characteristic acute diarrhea and weight loss of colitis. This study suggested that colostrums from sheep immunized against TNF-${\alpha}$ significantly inhibited TNF-${\alpha}$ bioactivity in the cell based assay. And the higher than anticipated variability in the two animal models precluded assessment of the ability of antibody to prevent TNF-${\alpha}$ induced intestinal damage in the intact animal. Further study will require to find out an alternative animal model, which is more acceptable to test anti-TNF-${\alpha}$ IgA therapy for reducing the impact of inflammation on gut dysfunction. And subsequent pre-clinical and clinical testing also need generation of more antibody as current supplies are low.

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The Role of c-Jun N-terminal Kinase in the Radiation-Induced Lung Fibrosis (방사선에 의한 폐 섬유화증에서 c-Jun N-terminal Kinase(JNK)의 역할)

  • Uh, Soo-Taek;Hong, Ki-Young;Lee, Young-Mok;Kim, Ki-Up;Kim, Do-Jin;Moon, Seung-Hyuk;Kim, Yong-Hoon;Park, Choon-Sik;Yeom, Uk;Kim, Eun-Suk;Choi, Doo-Ho
    • Tuberculosis and Respiratory Diseases
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    • v.50 no.4
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    • pp.450-461
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    • 2001
  • Background : The underlying pathogenesis of radiation-induced lung fibrosis (RTLF) has not been very well defined. However, the role of TGF-$\beta$ in the generation of RTLF has been a major focus because there is an increase in the expression of both the TGF-${\beta}m$-RNA and its protein preceding RTLF lesions. The down stream signal after a TGF-$\beta$ stimulated lung fibrosis includes the activation of many mediators such as Smad and c-Jun N-terminal kinase (JNK) through TAK1. It is we hypothesized that JNK activation may play a pivotal role in RTLF pathogenesis through increased transcription of the fibrogenic cytokines. The present study evaluates JNK activity in alveolar macrophages after irradiation and the relationship between JNK activity and the amount of collagen in the lung tissues. Methods : C57BL/6 mice(20-25 gr, males) received chlorotetracycline(2g/L) in their drinking water 1 week prior to irradiation and continuously there after. The mice were irradiated once with 1400 cGy of $60CO{\gamma}$-ray over the whole chest. The cellular composition of the whole lung bronchoalveoalr lavage fluids(BALF), elastin expression in the lung tissues, the level of hydroxyproline in lung tissues, and an in vitro JNK assay was measured before irradiation and one, four, and eight weeks after irradiation (RT). Results : The volumes of BALF retrieved from instilled 4 mL of saline with 2% heparin were 3.7-3.8 mL for each group. The cell numbers were similar before($4.1{\times}10^4{\pm}0.5{\times}10^4/mL$) and 1 week($3.1{\times}10^4{\pm}0.5{\times}10^4/mL$) after RT. At four and eight weeks after RT, the cell number reached to $14.0{\times}10^4{\pm}1.5{\times}10^4mL$ and $10.0{\times}10^4{\pm}1.3{\times}10^4/mL$, respectively. There we no changes in the lymphocytes and neutrophils population observed in the BALF after RT. The H-E stain of the lung tissues did not show any structural and fibrotic change in the lung tissues at 4 and 8 weeks after RT. In addition, the amount of elastin and collagen were not different on Verhoeff staining of the lung tissues before RT to eight weeks after RT. The hydroxyproine content was measured with the left lung dissected from the left main bronchus. The lung were homogenized and hydrolyzed with 6 N Hel for 12 hours at $110^{\circ}C$ then measured as previously described. The content of hydroxyproline, standardized with a lung protein concentration, reached a peak 4 weeks after RT, and thereafter showed a plateau. AnIn vitro JNK assay using c-$Jun_{1-79}$-GST sepharose beads were performed with the alveolar macrophages obtained from the BAL. JNK activity was not detected prior to RT, However, the JNK activity increased from one week after RT and reached a peak four weeks after RT. Conclusion : JNK may be involved in the pathogenesis because the JNK activity showed similar pattern observed with the hydroxyproine content. However, it is necessary to clarify that the JNK increases the transcription of fibrogenic cyiokines through the transcription factor.

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The Effect of Price Discount Rate According to Brand Loyalty on Consumer's Acquisition Value and Transaction Value (브랜드애호도에 따른 가격할인율의 차이가 소비자의 획득가치와 거래가치에 미치는 영향)

  • Kim, Young-Ei;Kim, Jae-Yeong;Shin, Chang-Nag
    • Journal of Global Scholars of Marketing Science
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    • v.17 no.4
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    • pp.247-269
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    • 2007
  • In recent years, one of the major reasons for the fierce competition amongst firms is that they strive to increase their own market shares and customer acquisition rate in the same market with similar and apparently undifferentiated products in terms of quality and perceived benefit. Because of this change in recent marketing environment, the differentiated after-sales service and diversified promotion strategies have become more important to gain competitive advantage. Price promotion is the favorite strategy that most retailers use to achieve short-term sales increase, induce consumer's brand switch, in troduce new product into market, and so forth. However, if marketers apply or copy an identical price promotion strategy without considering the characteristic differences in product and consumer preference, it will cause serious problems because discounted price itself could make people skeptical about product quality, and the changes of perceived value might appear differently depending on other factors such as consumer involvement or brand attitude. Previous studies showed that price promotion would certainly increase sales, and the discounted price compared to regular price would enhance the consumer's perceived values. On the other hand, discounted price itself could make people depreciate or skeptical about product quality, and reduce the consumers' positivity bias because consumers might be unsure whether the current price promotion is the retailer's best price offer. Moreover, we cannot say that discounted price absolutely enhances the consumer's perceived values regardless of product category and purchase situations. That is, the factors that affect consumers' value perceptions and buying behavior are so diverse in reality that the results of studies on the same dependent variable come out differently depending on what variable was used or how experiment conditions were designed. Majority of previous researches on the effect of price-comparison advertising have used consumers' buying behavior as dependent variable. In order to figure out consumers' buying behavior theoretically, analysis of value perceptions which influence buying intentions is needed. In addition, they did not combined the independent variables such as brand loyalty and price discount rate together. For this reason, this paper tried to examine the moderating effect of brand loyalty on relationship between the different levels of discounting rate and buyers' value perception. And we provided with theoretical and managerial implications that marketers need to consider such variables as product attributes, brand loyalty, and consumer involvement at the same time, and then establish a differentiated pricing strategy case by case in order to enhance consumer's perceived values properl. Three research concepts were used in our study and each concept based on past researches was defined. The perceived acquisition value in this study was defined as the perceived net gains associated with the products or services acquired. That is, the perceived acquisition value of the product will be positively influenced by the benefits buyers believe they are getting by acquiring and using the product, and negatively influenced by the money given up to acquire the product. And the perceived transaction value was defined as the perception of psychological satisfaction or pleasure obtained from taking advantage of the financial terms of the price deal. Lastly, the brand loyalty was defined as favorable attitude towards a purchased product. Thus, a consumer loyal to a brand has an emotional attachment to the brand or firm. Repeat purchasers continue to buy the same brand even though they do not have an emotional attachment to it. We assumed that if the degree of brand loyalty is high, the perceived acquisition value and the perceived transaction value will increase when higher discount rate is provided. But we found that there are no significant differences in values between two different discount rates as a result of empirical analysis. It means that price reduction did not affect consumer's brand choice significantly because the perceived sacrifice decreased only a little, and customers are satisfied with product's benefits when brand loyalty is high. From the result, we confirmed that consumers with high degree of brand loyalty to a specific product are less sensitive to price change. Thus, using price promotion strategy to merely expect sale increase is not recommendable. Instead of discounting price, marketers need to strengthen consumers' brand loyalty and maintain the skimming strategy. On the contrary, when the degree of brand loyalty is low, the perceived acquisition value and the perceived transaction value decreased significantly when higher discount rate is provided. Generally brands that are considered inferior might be able to draw attention away from the quality of the product by making consumers focus more on the sacrifice component of price. But considering the fact that consumers with low degree of brand loyalty are known to be unsatisfied with product's benefits and have relatively negative brand attitude, bigger price reduction offered in experiment condition of this paper made consumers depreciate product's quality and benefit more and more, and consumer's psychological perceived sacrifice increased while perceived values decreased accordingly. We infer that, in the case of inferior brand, a drastic price-cut or frequent price promotion may increase consumers' uncertainty about overall components of product. Therefore, it appears that reinforcing the augmented product such as after-sale service, delivery and giving credit which is one of the levels consisting of product would be more effective in reality. This will be better rather than competing with product that holds high brand loyalty by reducing sale price. Although this study tried to examine the moderating effect of brand loyalty on relationship between the different levels of discounting rate and buyers' value perception, there are several limitations. This study was conducted in controlled conditions where the high involvement product and two different levels of discount rate were applied. Given the presence of low involvement product, when both pieces of information are available, it is likely that the results we have reported here may have been different. Thus, this research results explain only the specific situation. Second, the sample selected in this study was university students in their twenties, so we cannot say that the results are firmly effective to all generations. Future research that manipulates the level of discount along with the consumer involvement might lead to a more robust understanding of the effects various discount rate. And, we used a cellular phone as a product stimulus, so it would be very interesting to analyze the result when the product stimulus is an intangible product such as service. It could be also valuable to analyze whether the change of perceived value affects consumers' final buying behavior positively or negatively.

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