• Title/Summary/Keyword: de novo

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Expression of $\beta$-amylase Gene and Degradation of Starch Granules of Germinating Rice Seed under Low Temperature and Submerged Soil Condition (저온.담수토양에서 벼종자 $\beta$-아밀라제 유전자 발현과 호분층 인접 배유의 전분분해 양상)

  • 윤병성;강원희
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.47 no.6
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    • pp.413-417
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    • 2002
  • This study was conducted to determine $\beta$-amylase gene expression and degradation of starch granules in the endosperm near scutellar epithelium of rice cultivars under the submerged soil at hypoxia 18$^{\circ}C$, which is practically important condition for farmers in temperate regions. In case of cv. Janghyangdo, accumulation of $\beta$-amylase mRNA was detected in the aleurone layer on the ninth day after seeding. However that of cv. Suwon 287 and Norm 6 were not detected in the aleurone layer in submerged soil(hypoxia) at 18$^{\circ}C$. $\beta$-amylase of cv. Janghyangdo was synthesized de novo in aleurone cells not in the scutellar epithelium. Degradation of starch granules in the endosperm near scutellar epithelium of c.v. Janghyangdo and Ginbozu, which have a strong $\beta$-amylase activity, was greater than that of cv. Suwon 287 and Norm 6 with no $\beta$-amylase activity in submerged soil(hypoxia) at 18$^{\circ}C$. This result may indicate that $\beta$-amylase gene expression and degradation of starch granules of germinating rice seed are related to the emergence of rice under the submerged soil condition at low temperature.

Increased mRNA Related Ovarian Maturation during Induction of Maturational Competence in Red Seabream, Pagrus major (참돔, Pagrus major의 성숙능력 유도시 증가된 난성숙 관련 mRNA)

  • Choi, Cheol-Young;Chang, Young-Jin;Takashima, Fumio
    • Development and Reproduction
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    • v.4 no.1
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    • pp.125-131
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    • 2000
  • This study has used differential display-PCR, to amplify genes transcribed during the ovarian maturation induced by human chorionic gonadotropin (hCG). The cDNA expressed at the times of acquisition of oocyte maturational competence in red seabream (Pagrus major) following treatment with hCG was amplified and cloned. A full-length of cDNA for p. major was isolated using differential display-PCR and 5'RACE. This cDNA clone contained 2,662 nucleotides including the open reading frame that encoded 434 amino acids. Homology analyses, using the GenBank and EMBL general database searches, indicated that the nucleotides sequence of the cDNA does not have high homology with any other genes. This cDNA was judged to be a gene, which induction of maturational competence coincides with increase of mRNA related ovarian maturation. Consensus sequences which were consistent with protein kinase C phosphorylation sites and casein kinase II phosphorylation sites were identified. in vitro, the transcription level of mRNA related ovarian maturation increased between 9hr and 24hr following treatment of ovarian follicles with hCG. It was also increased after GtH-II (300 ng/ml) stimulation. Furthermore, in vivo, mRNA related ovarian maturation was rarely expressed prior to the acquisition of oocyte maturational competence, but was strongly expressed after the acquisition of oocyte maturational competence, suggesting that the hCG induction of maturational competence is brought about by the de novo synthesis of the mRNA related ovarian maturation in p. major.

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Disruption of Sphingolipid Metabolism as a Potential Mechanism of Fumonisin Inhibition of Cell Growth in $LLC-PK_1$ Cells

  • Yoo, Hwan-Soo;Yun, Yeo-Pyo
    • Toxicological Research
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    • v.11 no.1
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    • pp.1-8
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    • 1995
  • Fumonisins are a family of mycotoxins produced by the fungus Fusarium moniliforme which is a common contaminant in corn. Fumonisins are potent inhibitors of sphingosine and sphinganine N-acyltransferase (ceramide synthase), key enzymes in sphingolipid metabolism. The purpose of this study was to provide the evidence that the elevated levels of free sphingoid bases (primarily sphinganine) and depletion of complex sphingolipids were closely related to the inhibition of cell growth in LLC-$PK_1$ cells exposed to fumonisin $B_1$$(\leq 35 {\mu}M)$. Concentrations of fumonisin $B_1$ between 10 and $35 {\mu}M$ were known to inhibit cell growth without cytotoxicity in $LLC-PK_1$ cells (Yoo et al. Toxicol. Appl. Pharmacol. 114, 9-15, 1992). Cells exposed to 35$\mu M$ fumonisin B$_1$ for 48 and 72 hr developed a fibroblast-like (elongated and spindle-shaped) appearance and were less confluent than normal cells. At between 24 and 48 hr after exposure to fumonisin $B_1$ cells were beginning to show the inhibition of cell growth and at 72 hr the number of viable cells in fumonisin-treated cultures was about 50% of concurrent control cultures. During the 24 hr lag period preceding inhibition of cell growth, the free sphinganine levels in cells exposed to $35 {\mu}M$ fumonisin $B_1$ were highly elevated (approximately 230 fold higher than normal cells). The elevated levels of free sphinganine were $435\pm14$$pmoles/{10^6}$ cells at 48 hr and approximately TEX>$333\pm11$$pmoles/{10^6}$ cells in cells exposed to $35{\mu}M$ fumonisin$B_1$ at 72 hr, while the levels of free sphinganine in normal cells were less than 2$pmoles/{10^6}$ cells. Under the same condition, depletion of intracellular complex sphingolipids as a consequence of fumonisin inhibition of de novo sphingolipid biosynthesis and turnover pathway was appeared. Content of free sphingold bases in dividing cells was more elevated than in confluent cells at 24-48 hr after cells were exposed to $20{\mu}M$ fumonisin $B_1$. The dividing cells were showing the inhibition of cell growth at 48-72 hr and $20{\mu}M$ fumonisin $B_1$. The results of this study support the hypothesis that the inhibition of cell growth is very well related to the disruption of sphingolipid metabolism in $LLC-PK_1$ cells.

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Growth Temperature-Dependent Conversion of De novo-Synthesized Unsaturated Fatty Acids into Polyhydroxyalkanoic Acid and Membrane Cyclopropane Fatty Acids in the Psychrotrophic Bacterium Pseudomonas fluorescens BM07

  • LEE , HO-JOO;RHO, JONG-KOOK;YOON, SUNG-CHUL
    • Journal of Microbiology and Biotechnology
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    • v.14 no.6
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    • pp.1217-1226
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    • 2004
  • A psychrotrophic bacterial strain, Pseudomonas fluorescens BM07, synthesized unsaturated fatty acids (UFA) from fructose in response to lowering of growth temperature, and incorporated them into both polyhydroxyalkanoic acid (PHA) and membrane lipid. The blocking of PHA synthesis by adding 5 mM 2-bromooctanoic acid to the growth medium, containing 70 mM fructose, was found to be a useful means to profile the composition of membrane lipid by gas chromatography. As the growth temperature changed from 35 to $50^{\circ}C$, the total content of two UFA, 3-hydroxy-cis-5­dodecenoic acid ($C_{12:1}$) and 3-hydroxy-cis-7-tetradecenoic acid ($C_{14:1}$), in PHA increased from 31 to 44 $mol\%$. The growth at lower temperatures also led to an increase in the level of two major UFA, palmitoleic acid (C16:1 cis9) and cis-vaccenic acid (C18:1 cis11), in membrane lipid. A fraction of these membrane-lipid UFA was converted to their corresponding cyclopropane fatty acids (CFA). The CFA conversion was a function of culture time, exhibiting biphasic increase before and after entering the stationary phase. However, pH changes in growth media had no effect on the CFA conversion, which is contrary to the case of E. coli reported. The cells grown at $30^{\circ}C$ responded to a cold shock (lowering the medium temperature down to $10^{\circ}C$) by increasing the level of C16:1 cis9 and C 18: I cis II up to that of $10^{\circ}C$-grown control cells and concomitantly decreasing the relative level of cis-9,10­methylenehexadecanoic acid (the CFA converted from C16:1 cis9) from 14 to 8 $mol\%$, whereas the 10-grown cells exhibited little change in the lipid composition when exposed to a warmer environment of $30^{\circ}C$ for 12 h. Based on this one- way response, we suggest that this psychrotrophic strain responds more efficiently and sensitively to a cold shock than to a hot shock. It is also suggested that BM07 strain is a good producer of two unsaturated 3-hydroxyacids, $C_{12:1}\;and\;C_{141:1}$.

Sodium Dependent Taurine Transport into the Choroid Plexus, the Blood-Cerebrospinal Fluid Barrier

  • Chung, Suk-Jae;Ramanathan, Vikram;Brett, Claire M.;Giacomini, Kathleen M.
    • Journal of Pharmaceutical Investigation
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    • v.25 no.3
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    • pp.7-20
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    • 1995
  • Taurine, a ${\beta}-amino$ acid, plays an important role as a neuromodulator and is necessary for the normal development of the brain. Since de novo synthesis of taurine in the brain is minimal and in vivo studies suggest that taurine dose not cross the blood-brain barrier, we examined whether the choroid plexus, the blood-cerebrospinal fluid (CSF) barrier, plays a role in taurine transport in the central nervous system. The uptake of $[^3H]-taurine$ into ATP depleted choroid plexus from rabbit was substantially greater in the presence of an inwardly directed $Na^+$ gradient taurine accumulation was negligible. A transient in side-negative potential gradient enhanced the $Na^+-driven$ uptake of taurine into the tissue slices, suggesting that the transport process is electrogenic, $Na^+-driven$ taurine uptake was saturable with an estimated $V_{max}$ of $111\;{\pm}\;20.2\;nmole/g/15\;min$ and a $K_M\;of\;99.8{\pm}29.9\;{\mu}M$. The estimated coupling ratio of $Na^+$ and taurine was $1.80\;{\pm}\;0.122.$ $Na^+-dependent$ taurine uptake was significantly inhibited by ${\beta}-amino$ acids, but not by ${\alpha}-amino$ acids, indicating that the transporter is selective for ${\beta}-amino$ acids. Since it is known that the physiological concentration of taurine in the CSF is lower than that in the plasma, the active transport system we characterized may face the brush border (i.e., CSF facing) side of the choroid plexus and actively transport taurine out of the CSF. Therefore, we examined in vivo elimination of taurine from the CSF in the rat to determine whether elimination kinetics of taurine from the CSF is consistent with the in vitro study. Using a stereotaxic device, cannulaes were placed into the lateral ventricle and the cisterna magna of the rat. Radio-labelled taurine and inulin (a marker of CSF flow) were injected into the lateral ventricle, and the concentrations of the labelled compounds in the CSF were monitored for upto 3 hrs in the cisterna magna. The apparent clearance of taurine from CSF was greater than the estimated CSF flow (p<0.005) indicating that there is a clearance process in addition to the CSF flow. Taurine distribution into the choroid plexus was at least 10 fold higher than that found in other brain areas (e. g., cerebellum, olfactory bulb and cortex). When unlabelled taurine was co-administered with radio-labelled taurine, the apparent clearance of taurine was reduced (p<0.0l), suggesting a saturable disposition of taurine from CSF. Distribution of taurine into the choroid plexus, cerebellum, olfactory bulb and cortex was similarly diminished, indicating that the saturable uptake of taurine into these tissues is responsible for the non-linear disposition. A pharmacokinetic model involving first order elimination and saturable distribution described these data adequately. The Michaelis-Menten rate constant estimated from in vivo elimination study is similar to that obtained in the in vitro uptake experiment. Collectively, our results demonstrate that taurine is transported in the choroid plexus via a $Na^+-dependent,saturable$ and apparently ${\beta}-amino$ acid selective mechanism. This process may be functionally relevant to taurine homeostasis in the brain.

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Systematic review of the effect of omega-3 fatty acids on improvement of blood flow while focused on evaluation of claims for health functional food (건강기능식품의 기능성을 중심으로 한 오메가-3 지방산 함유유지의혈행개선 효과에 대한 체계적 고찰)

  • Jeong, Sewon;Kim, Ji Yeon;Paek, Ju Eun;Kim, Joohee;Kwak, Jin Sook;Kwon, Oran
    • Journal of Nutrition and Health
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    • v.46 no.3
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    • pp.226-238
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    • 2013
  • Omega-3 polyunsaturated fatty acids are essential fatty acids because humans cannot synthesize them de novo and must obtain them in their diet. Fish and fish oil are rich sources of omega-3 fatty acids, including eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). Significant evidence of the beneficial role of dietary intake of omega-3 fatty acids in blood flow has been reported and putative mechanisms for improvement of blood flow include anti-thrombotic effects, lowered blood pressure, improved endothelial function, and anti-atherogenic effects. Edible oils containing omega-3 fatty acids were registered as functional ingredients in the Korea Health Functional Food Code. Although omega-3 fatty acids have been evaluated by the Korea Food and Drug Administration (KFDA) based on scientific evidence, periodic re-evaluation may be needed because emerging data related to omega-3 fatty acids have accumulated. Therefore, in this study, we re-evaluated scientific evidence for the effect of omega-3 fatty acids as a functional ingredient in health functional food on improvement of blood flow. A comprehensive literature search was conducted for collection of relevant human studies using the Medline and Cochrane, KISS, and IBIDS databases for the years 1955-2012. Search keywords were used by combination of terms related to omega-3 fatty acids and blood flow. The search was limited to human studies published in Korean, English, and Japanese. Using the KFDA's evidence based evaluation system for scientific evaluation of health claims, 112 human studies were identified and reviewed in order to evaluate the strength of the evidence supporting a relation between omega-3 fatty acids and blood flow. Among 112 studies, significant effects on improvement of blood flow were reported in 84 studies and the daily intake amount was ranged from 0.1 to 15 g. According to this methodology of systematic review, we concluded that there was possible evidence to support a relation between omega-3 fatty acid intake and blood flow. However, because inconsistent results have recently been reported, future studies should be monitored.

Role of the MDM2 Promoter Polymorphism (-309T>G) in Acute Myeloid Leukemia Development

  • Cingeetham, Anuradha;Vuree, Sugunakar;Jiwatani, Sangeeta;Kagita, Sailaja;Dunna, Nageswara Rao;Meka, Phanni Bhushann;Gorre, Manjula;Annamaneni, Sandhya;Digumarti, Raghunadharao;Sinha, Sudha;Satti, Vishnupriya
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.7
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    • pp.2707-2712
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    • 2015
  • Background: The human homologue of the mouse double minute 2 (MDM2) gene is a negative regulator of Tp53. MDM2-309T>G a functional promoter polymorphism was found to be associated with overexpression thereby attenuation of Tp53 stress response and increased cancer susceptibility. We have planned to evaluate the possible role of MDM2-309T>G polymorphism with risk and response to chemotherapy in AML. Materials and Methods: A total of 223 de novo AML cases and 304 age and sex matched healthy controls were genotyped for the MDM2-309T>G polymorphism through the tetra-primer amplification refractory mutation system (ARMS)-PCR method. In order to assess the functional relationship of -309T>G SNP with MDM2 expression level, we quantified MDM2 mRNA in 30 primary AML blood samples through quantitative RT-PCR. Both the (-309T>G) genotypes and the MDM2 expression were correlated with disease free survival (DFS) rates among patients who have achieved complete remission (CR) after first induction chemotherapy. Results: MDM2-309T>G polymorphism was significantly associated with AML development (p<0.0001). The presence of either GG genotype or G allele at MDM2-309 confered 1.79 (95% CI: 1.12-2.86; p<0.001) and 1.46 fold (95%CI: 1.14-1.86; p= 0.003) increased AML risk. Survival analysis revealed that CR+ve cases with GG genotype had significantly increased DFS rates (16months, p=0.05) compared to CR+ve TT (11 months) and TG (9 months) genotype groups. Further, MDM2 expression was also found to be significantly elevated in GG genotype patients (p=0.0039) and among CR+ve cases (p=0.0036). Conclusions: The MDM2-309T>G polymorphism might be involved in AML development and also serve as a good prognostic indicator.

Expression Pattern of the Trefoil Factor Family 1 in Gastric Adenoma and Carcinoma (위 선종 및 선암에서 Trefoil Factor Family 1 단백의 발현 양상)

  • Park Won Sang;Kim Young Sil;Yoo Nam Jin;Park Cho Hyun;Yoo Jin Young;Lee Youn Soo;Lee Jung Young
    • Journal of Gastric Cancer
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    • v.1 no.1
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    • pp.4-9
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    • 2001
  • Purpose: The trefoil factor family 1 (TFF1) has a protective effect against gastric mucosal damage induced by nonsteroidal anti-inflammatory drugs or ethanol. In addition, a TFF1 knockout mouse model has exhibited circumferential adenomas with high-grade dysplasia, of which $30\%$ progressed into frankly invasive carcinomas. We tried to determine whether the expression pattern of the TFF1 could be involved in the development of sporadic gastric carcinomas. Materials and Methods: We examined TFF1 expression in a series of 43 sporadic gastric carcinomas and 18 gastric adenomas by immunohistochemistry. Results: Strong positive TFF1 staining was identified primarily in the normal gastric mucosa, mainly in the cytoplasm of the superficial and foveolar epithelium. We found TFF1 expression in $55.8\%$ (24 out of 43) of the gastric carcinomas and in $16.7\%$ (3 out of 18) of the gastric adenomas. Statistically, TFF1 immunoreactivity was significantly higher in diffuse-type ($82.4\%$) than in intestinal-type ($38.5\%$) carcinomas(p=0.0058, Fisher's exact test). Conclusion: Our findings provide sufficient evidence that the expression of TFF1 in gastric cancer may simply disclose gastric-type differentiation of neoplastic cells and provide further support for the existence of at least two pathways of malignant transformation of the gastric mucosa: one via intestinal metaplasia and adenomatous dysplasia, leading to glandular carcinomas with intestinal-type differentiation, and the other via hyperplastic changes or de novo changes, leading to diffuse carcinomas and to a subset of glandular carcinomas displaying gastric-type differentiation.

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Transcriptome Profiling and Characterization of Drought-Tolerant Potato Plant (Solanum tuberosum L.)

  • Moon, Ki-Beom;Ahn, Dong-Joo;Park, Ji-Sun;Jung, Won Yong;Cho, Hye Sun;Kim, Hye-Ran;Jeon, Jae-Heung;Park, Youn-il;Kim, Hyun-Soon
    • Molecules and Cells
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    • v.41 no.11
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    • pp.979-992
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    • 2018
  • Potato (Solanum tuberosum L.) is the third most important food crop, and breeding drought-tolerant varieties is vital research goal. However, detailed molecular mechanisms in response to drought stress in potatoes are not well known. In this study, we developed EMS-mutagenized potatoes that showed significant tolerance to drought stress compared to the wild-type (WT) 'Desiree' cultivar. In addition, changes to transcripts as a result of drought stress in WT and drought-tolerant (DR) plants were investigated by de novo assembly using the Illumina platform. One-week-old WT and DR plants were treated with -1.8 Mpa polyethylene glycol-8000, and total RNA was prepared from plants harvested at 0, 6, 12, 24, and 48 h for subsequent RNA sequencing. In total, 61,100 transcripts and 5,118 differentially expressed genes (DEGs) displaying up- or down-regulation were identified in pairwise comparisons of WT and DR plants following drought conditions. Transcriptome profiling showed the number of DEGs with up-regulation and down-regulation at 909, 977, 1181, 1225 and 826 between WT and DR plants at 0, 6, 12, 24, and 48 h, respectively. Results of KEGG enrichment showed that the drought tolerance mechanism of the DR plant can mainly be explained by two aspects, the 'photosynthetic-antenna protein' and 'protein processing of the endoplasmic reticulum'. We also divided eight expression patterns in four pairwise comparisons of DR plants (DR0 vs DR6, DR12, DR24, DR48) under PEG treatment. Our comprehensive transcriptome data will further enhance our understanding of the mechanisms regulating drought tolerance in tetraploid potato cultivars.

The Properties of Rice Flours Prepared by Dry- and Wet-Milling of Soaked Glutinous and Normal Grains (수침과 건조조건을 달리하여 만든 찹쌀가루와 멥쌀가루의 특성)

  • Kim, Wan-Soo;Shin, Mal-Shick
    • Korean journal of food and cookery science
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    • v.23 no.6
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    • pp.908-918
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    • 2007
  • To prepare ready-to-use rice flour as de novo material for processed rice foods, glutinous(W) and normal rice grains (N) were soaked for 1, 8, and 12 hours prior to processing. One half (DG) was air-dried and milled, and the other (WG) was milled and air-dried. General, morphological and pasting properties of the flours (NDG, NWG, WDG, WWG) were compared to those of a control (raw milled rice without soaking). The general compositions of the rice flours varied with soaking. Crude ash was considerably decreased at the beginning of soaking (1 hour). With the soaking, the rice flour, having polygonal shaped particles and a layered surface, acquired particles with smooth edges, which were then uniformly distributed. Additionally, the WG flour was lighter and had a lower ${\Delta}E$ value than the DG flour, due to a higher L and less +b as a result of soaking. Compared to the control, the WBC of the normal rice flour was decreased significantly with soaking, and the WG flour had significantly lower WBCs than the DG flour. Stirring number (SN), an indicator of ${\alpha}-amylase$ activity, was highly and significantly correlated with WBC (r=-0.85, p=0.0001) in the normal rice flour. At $80^{\circ}C$, the SP and solubility of all the soaked rice flours were much higher than those of the control. Positive (r=+0.85, p=0.0001) and negative (r=-0.61, p=0.02) correlations between the SP and solubility of the normal and glutinous rice flours were found, respectively. Using RVA, the pasting temperature of NDG was lower than that of NWG (p<0.0001). The peak viscosities of all the soaked flours were significantly decreased with soaking (p<0.0l), with the highest viscosity in the normal rice flour soaked for 8 hrs. Total setback, indicative of retrogradation, was lower in NDG than in NWG, with the lowest setback at 8 hrs of' soaking. Based on these finding, the NDG flour with 8 hrs of soaking was less damaged, and had a lower total setback and lower pasting temperature, which would make it an appropriate rice flour for commercial mass production.