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Development of EvaGreen Based Real-time PCR Assay for Detection and Quantification Toxic Dinoflagellate Pfiesteria Piscicida and Field Applications (유독 와편모조류 Pfiesteria Piscicida 탐지 및 정량 분석을 위한 EvaGreen 기반 Real-time PCR기법 개발과 현장 적용)

  • PARK, BUM SOO;JOO, JAE-HYOUNG;KIM, MYO-KYUNG;KIM, JOO-HWAN;KIM, JIN HO;BAEK, SEUNG HO;HAN, MYUNG-SOO
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.22 no.1
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    • pp.31-44
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    • 2017
  • Pfiesteria piscicida is one of heterotrophic dinoflagellate having toxic metaboliges, and it is difficult to detect and quantify this dinoflagellate via light microscope due to small size and morphological similarity with Pfiesteria-like dinoflagellate (PLD) species. Alternatively, we developed quantitative real-time PCR assay based on EvaGreen and determined field accessibility throughout the investigation of distribution in the entire Korean coastal waters and population dynamics in Shihwa Lake. The P. piscicida-specific primers based on internal transcribed spacer 1 (ITS 1) were designed and the specificity of primers was confirmed by PCR with other genomic DNAs which have genetic similarity with target species. Through real-time PCR assay, a standard curve which had a significant linear correlation between log cell number and $C_T$ value ($r^2{\geq}0.999$) and one informative melting peak ($88^{\circ}C$) were obtained. These results implies that developed real-time PCR can accurately detect and quantify P. piscicida. Throughout the field applications of real-time PCR assay, P. piscicida was distributed in western (Mokpo and Kimje) and easthern (Gangneng) Korean coastal water even though light microscopy failed to identify P. piscicida. In the investigation of population dynamics in Shihwa Lake, the density of P. piscicida was peaked in June, July and August 2007 at St. 1 where salinity (${\leq}15psu$) was lower than the other 2 sites. In this study, we successed to develop EvaGreen bassed real-time PCR for detection and quantification of P. piscicida in fields, so this developed assay will be useful for various ecological studies in the future.

Characteristics of the Differences between Significant Wave Height at Ieodo Ocean Research Station and Satellite Altimeter-measured Data over a Decade (2004~2016) (이어도 해양과학기지 관측 파고와 인공위성 관측 유의파고 차이의 특성 연구 (2004~2016))

  • WOO, HYE-JIN;PARK, KYUNG-AE;BYUN, DO-SEONG;LEE, JOOYOUNG;LEE, EUNIL
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.23 no.1
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    • pp.1-19
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    • 2018
  • In order to compare significant wave height (SWH) data from multi-satellites (GFO, Jason-1, Envisat, Jason-2, Cryosat-2, SARAL) and SWH measurements from Ieodo Ocean Research Station (IORS), we constructed a 12 year matchup database between satellite and IORS measurements from December 2004 to May 2016. The satellite SWH showed a root mean square error (RMSE) of about 0.34 m and a positive bias of 0.17 m with respect to the IORS wave height. The satellite data and IORS wave height data did not show any specific seasonal variations or interannual variability, which confirmed the consistency of satellite data. The effect of the wind field on the difference of the SWH data between satellite and IORS was investigated. As a result, a similar result was observed in which a positive biases of about 0.17 m occurred on all satellites. In order to understand the effects of topography and the influence of the construction structures of IORS on the SWH differences, we investigated the directional dependency of differences of wave height, however, no statistically significant characteristics of the differences were revealed. As a result of analyzing the characteristics of the error as a function of the distance between the satellite and the IORS, the biases are almost constant about 0.14 m regardless of the distance. By contrast, the amplitude of the SWH differences, the maximum value minus the minimum value at a given distance range, was found to increase linearly as the distance was increased. On the other hand, as a result of the accuracy evaluation of the satellite SWH from the Donghae marine meteorological buoy of Korea Meteorological Administration, the satellite SWH presented a relatively small RMSE of about 0.27 m and no specific characteristics of bias such as the validation results at IORS. In this paper, we propose a conversion formula to correct the significant wave data of IORS with the satellite SWH data. In addition, this study emphasizes that the reliability of data should be prioritized to be extensively utilized and presents specific methods and strategies in order to upgrade the IORS as an international world-wide marine observation site.

A Comparative Study of Blood Lead Measurement by Polarized Zeman Effect Correction AAS and D2 Correction AAS Method (편광 Zeeman 보정 및 D2 보정 방법에 의한 혈중연 측정치의 비교 연구)

  • Lee, Seok Ki;Ahn, Kyu Dong;Lee, Byung Kook
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.5 no.1
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    • pp.59-67
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    • 1995
  • Blood lead assay by $D_2$ lamp background correction method of atomic absorption spectrophotometer(AAS) with wavelength of 283.3 nm is most popular in occupational health practice in Korea. On the other hand, $D_2$ lamp background correction method with wavelength of 217.0 nm is also often used in general chemical analysis for lead assay in general purpose. But both methods have some weakness of background correction which brought direct effect on the results of analysis. Recently blood lead assay with polarized Zeeman effect of AAS was introduced and is now preferred in many laboratory than $D_2$ correction method in blood lead analysis. But still AAS with $D_2$ lamp are widely used in the field of occupational health in Korea. This study compared blood lead assay data with $D_2$ correction methods(283.3 and 217.0 nm) and with that of polarized Zeeman effect correction method to evaluate the validity of 02 correction methods. The results obtained were as follows; 1. Taking the value of polarized Zeeman effect method as reference value of 1.00, the mean relative value of $D_2$ correction method with wavelength of 217.0 nm was 0.92 and that with wavelength of 283.3 nm was 0.90 respectively in the analysis of blood lead whose value were below $20.0{\mu}g/dl$(p<0.001). Both mean values were statistically smaller than polarized Zeeman effect correction method. But in the analysis of blood whose value were between 20.0 to $20.0{\mu}g/dl$, the mean relative value of $D_2$ correction method was 0.96 in both wavelength and did not differ from polarized Zeeman effect method(p<0.001). There was no difference of blood lead between $D_2$ correction method and polarized Zeeman effect method in the analysis of blood lead whose value were over $40.0{\mu}g/dl$. 2. The variations of background correction value in polarized Zeeman effect method were not changed by increase of blood lead, but those in $D_2$ correction methods were increased by the increase of blood lead. While then relative standard deviation(RSD) of data measured by Zeeman effect method were decreased by the increase of blood lead, those by $D_2$ methods were nol differed by the increase of blood lead.

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Climate-related Changes in Fruit Growth of 'Fuyu' Persimmon during the Harvest Season (수확기 동안의 기상 변화에 따른 '부유' 감의 과실 생장)

  • Choi, Seong-Tae;Park, Doo-Sang;Son, Ji-Young;Park, Yeo-Ok;Hong, Kwang-Pyo;Cho, Kwang-Sik
    • Horticultural Science & Technology
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    • v.31 no.1
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    • pp.32-37
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    • 2013
  • Relationships among climate changes, early frost, and fruit growth were studied during the final month to harvest of late-maturing 'Fuyu' persimmon (Diospyros kaki) to assess the changes in fruit characteristics during this critical period. The heavy frost on Nov. 16 defoliated more than 70% of the leaves, but with little damage on the fruits. However, all the leaves were defoliated by the heavy frost on Nov. 20, and all the fruits were cold-damaged by $-3.3^{\circ}C$ on Nov. 21. Fruit weight increased by 8-25 g per week from Oct. 25 (142 days after full bloom) to Nov. 15, reaching to 250 g, but it decreased by 3-4 g per week after the frost. Hunter a value of fruit skin gradually increased until the last harvest on Nov. 29 with a temporary halt in early Nov. when temperature was high, whereas fruit firmness rapidly decreased after the frost on Nov. 21. Fruit soluble solids were $15.7-16.1^{\circ}Brix$ for the final month. When some branches were covered with non-woven fabrics to avoid direct contact with frost, the fruits on the branches were not visually damaged by the low temperature although 40-60% of their leaves were defoliated on Nov. 16. However, low temperature on Nov. 20 and 21 defoliated all the leaves, causing cold damage on the fruits. There was a highly significant correlation between the fruit diameter and its weight ($R^2$ = 0.73-0.91). So, the regression equations could be used to estimate weight from diameter of the fruits sampled from the branches with the non-woven fabrics. The calculated fruit weight reached to a maximum of 240 g on Nov. 15. Daily increases in fruit weight were 1.1-2.5 g from Oct. 25 to 31, 1.9-3.5 g from Oct. 31 to Nov. 7, and 1.4-1.6 g from Nov. 7 to 12. However, fruit weight decreased by 0.3-1 g per day after the cold damage on Nov. 21. The results indicate that the most appropriate harvest time could be dependent on relationship of fruit growth to climate.

Relationship between Limnological Characteristics and Algal Bloom in Lake-type and River-Type Reservoirs, Korea (호소형 및 하천형 댐 호의 육수학적 특성과 조류발생과의 상관관계)

  • Kim, Jong-Min;Heo, Seong-Nam;Noh, Hye-Ran;Yang, Hee-Jeong;Han, Myung-Soo
    • Korean Journal of Ecology and Environment
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    • v.36 no.2 s.103
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    • pp.124-138
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    • 2003
  • This paper aimed to analyze the relationship between alga3 bloom patterns and hydrological, limnological data which were collected from major reservoirs in Korea for 8 years (1990${\sim}$1997). Water temperature of river-type reservoirs showed wider seasonal fluctuations than that of lake-type. pH of lake-type reservoirs was low in winter season but high in summer season. In contrast, river-type reservoirs showed high pH in spring and autumn seasons as well, and very low in summer season. COD of lake-type reservoirs and Paldang reservoir was lower (2${\sim}$3 mg/L) than that of Geumgang and Nagdonggang reservoirs (6${\sim}$9 mg/L). Dissolved oxygen (DO) of river-type reservoirs was higher than that of lake-type reservoirs. Seasonal fluctuation pattern of DO saturation in river-type reservoirs was high (80 ${\sim}$100%) and remained relatively constant whereas lake-type reservoirs showed the highest level (93%) in late spring or early summer, which gradually decreased entering winter season(46${\sim}$06%). And monthly variation of DO saturation showed inverse proportion to water volume in lake-type reservoirs. Nutrients concentration in river-type lake is higher than lake-type. Seasonal fluctuation of nutrients (T-N, T-P) in lake-type reservoirs was relatively small than that of river-type reservoirs. Annual mean N/P mass ratio of lake-type reservoirs was higher than that of river-type. Transparency tended to related with the suspended solid concentration in river-type reservoirs. Algal bloom of lake-type and river-type reservoirs occurred at any time except rainfall and winter periods. And it dominated in summer and early autumn, respectively. Algal bloom of river-type reservoirs was higher than that of lake-type. Relationship between rainfall and chlorophyll- a in lake-type reservoirs was relatively high, however river-type reservoirs showed insignificant.

Wintering Population Change of the Cranes according to the Climatic Factors in Cheorwon, Korea: Effect of the Snow Cover Range and Period by Using MODIS Satellite Data (기후요인에 의한 철원지역 두루미류 월동개체수 변화 - MODIS 위성영상을 이용한 눈 덮임 범위와 지속기간의 영향 -)

  • Yoo, Seung-Hwa;Lee, Ki-Sup;Jung, Hwa-Young;Kim, Hwa-Jung;Hur, Wee-Haeng;Kim, Jin-Han;Park, Chong-Hwa
    • Korean Journal of Ecology and Environment
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    • v.48 no.3
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    • pp.176-187
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    • 2015
  • In this study, we hypothesized that the size of wintering crane population would change due to the climate factors. We assumed that wintering population size would differ by climate values in January, which is the coldest period in year. Especially, White-naped cranes were able to choose wintering site between Cheorwon and other alternative place where snow coverage had low influence, differing from Red crowned cranes. For this reason, we predicted the population size of White-naped cranes would fluctuate according to the extent of snow coverage in Cheorwon. Therefore we used snow coverage data based on MODIS and climate data from KMA (Korea Meteorological Administration) that are generally used. We analyzed the crane's population size in Cheorwon in January from 2002 to 2014. The temperature in the Cheorwon increased from 2002 to wintering period in 2007~ 2008 and went down, showing the lowest temperature in 2011~ 2012. With this phenomenon, warmth index showed the similar pattern with temperature. Amount of newly accumulated snow (the amount of snow that fallen from 0:01 am to 11:29 pm in a day) was low after 2002, but rapidly increased in 2010~ 2011 and 2011~ 2012. The area of snow coverage rapidly declined from 2002 to 2005~ 2006 but suddenly expanded in wintering period in 2009~ 2010 and 2010~ 2011. Wintering population size of the White-naped cranes decreased as snow coverage area increased in January and the highest correlation was found between them, compared to the other climatic factors. However, the number of individuals of Red crowned cranes had little relationship with general climate factors including snow cover range. Therefore it seems that population size of the Red crowned crane varied by factors related with habitat selection such as secure roosting site and area of foraging place, not by climatic factors. In multiple regression analysis, wintering population of White-naped cranes showed significant relationship with logarithmic value of snow cover range and its period. Therefore, it suggests that the population size of the White-naped crane was affected by snow cover range n wintering period and this was because it was hard for them to find out rice grains which are their main food items, buried in snow cover. The population size variation in White-naped cranes was caused by some individuals which left Cheorwon for Izumi where snow cover had little influence on them. The wintering population in Izumi and Cheorwon had negative correlation, implying they were mutually related.

A Study of Fish Community on Up and Downstream of Hwabuk Dam Under Construction in the Upper Wie Stream. (위천 상류에 건설 중인 화북댐 상 하류 어류군집에 관한 연구)

  • Seo, Jin-Won;Kim, Hee-Sung
    • Korean Journal of Ecology and Environment
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    • v.42 no.2
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    • pp.260-269
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    • 2009
  • Hwabuk Dam has been under construction to reduce flood damage in Nakdong River watershed and to supply stable water for middle area of Gyeongbuk Province. Therefore, fish investigation in up and downstream of the dam was conducted from 2004 to 2008 in order to determine any negative effect on fish community due to dam construction and to use as fundamental data for conserving species diversity and maintaining stream health. According to data analysis on water quality, temperature, dissolved oxygen, pH, suspended solids, and total E-coli had seasonal variation, but they did not significantly differ in sites. However, biological and chemical oxygen demand, chlorophyll-a, nitrogen, and phosphorus representing organic matter and nutrient concentration were higher in upper site and decreased to lower site so that they differed by site. Concentration of arsenic among the heavy metals was less than 0.05 mg $L^{-1}$, which is regulated for protection of human health in water quality standard, except for 0.092 mg $L^{-1}$ in June 2005. During the study period, the total number of fish caught from the 6 sites was 10,263 representing 7 families 19 species. Among them, dominant and subdominant species were Korean chub (Zacco koreanus, 62.5%) and Chinese minnow (Rhynchocypris oxycephalus, 10.6%) which inhabit mostly in mid and upper streams, Korea. Among the 19 species, Korean endemic species were 9 species (47.4%) including Korean slender gudgeon (Squalidus gracilis majimae), Korean dark sleeper (Odontobutis platycephala), and Korean shiner (Coreoleuciscus splendidus). There was several individuals of the $1^{st}$-class endangered species, Naktong nose loach (Koreocobitis nahtongensis), caught in 2005${\sim}$2007, and no introduced species of fish was found in entire sampling period. According to result of community analysis, dominance index decreased toward lower site, but diversity and richness indices increased toward lower site. The equation of length-weight relationship on the dominant species was TW=0.000003$(TL)^{3.2603}$. The parameter b in the equation was greater than 3.0 indicating good nutritional condition in the populations. Compared to populations of Korean chub in other streams, the population in Hwabuk Dam watershed had higher mean of condition factor by size indicating better growth rate. With fish fauna and multi-metric health assessment model in each sampling attempt, index of biotic integrity (IBI) was evaluated and it resulted mostly in good (26${\sim}$35) and excellent (36${\sim}$40) condition in all sites, and the mean of IBI was the highest in site 5. The results indicate that it is very important to study not only environmental impact assessment with fish composition but also stream health assessment in order to conserve healthy aquatic ecosystem.

Effects of TDN/CP Ratio on Nutrient Intake, Body Weight Gain, Blood Characteristics of Dairy Heifers from 12 to 14 Months of Age (TDN/CP 비율이 젖소 12~14개월령 육성우 영양소 섭취량, 증체량 및 혈액성상에 미치는 영향)

  • Kim, Gyeom-Heon;Hwang, Won-Uk;Kim, Hyun-Jin;Kim, Soo-Ki
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.36 no.2
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    • pp.115-123
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    • 2016
  • In this study, the effect of energy and protein ratio on the nutrient availability and body weight gain was determined to find the proper nutrient level for the stage of replacement dairy heifers before fertilization at approximately 12~24 months old after birth. Dry matter intake showed that the TDN/CP rate of T1 (5.0:1), T2 (4.5:1) and T3 (4.0:1) resulted in similar values. Among feces, the ash content of T1 was 11.07%, which was significantly (p < 0.05) higher than that of T3 (9.69%). It was also significantly (p < 0.05) higher than that of T2 (10.32%). When the TDN/CP rate was increased, the ash content in the feces was also increased. Regardless of the initial weight, weight gain was increased significantly as the CP rate in the feed was increased on the $30^{th}$ day. On the $60^{th}$ day, T2 showed the highest rate of gain (p < 0.05). As the TDN rate was decreased or the protein rate was increased, weight gain was significantly (p < 0.05) increased. Regarding GPT level in the blood after finishing the test, T1 and T3 (112.6, 88.3 u/l) showed significantly (p < 0.05) higher value than T2 (50.9 u/l). For phosphorus level, T1 (46.3 mg/dl) showed significantly (p < 0.05) higher value than other treatments. For HDL cholesterol level, T2 (145.2 mg/dl) had significantly (p < 0.05) higher level than T1 (121.0 mg/dl) or T3 (132.3 mg/dl). For triglyceride levels, T3 (40.6 mg/dl) had significantly (p < 0.05) higher value than T1 (20.7 mg/dl) and T2 (29.0 mg/dl). For other blood parameters including BUN concentration, there were no significant (p > 0.05) difference among treatments. As a result, although feed intake and weight gain with TDN/CP rate of 4:1 showed best results, considering the excess body fat accumulation possibilities or blood metabolism, it seems 4.5:1 ratio is most appropriate.

Preventive Effects of Rosa rugosa Root Extract on Advanced Glycation End product-Induced Endothelial Dysfunction (해당근 추출물의 항산화 활성 및 최종당화산물에 의한 혈관내피세포 기능장애 억제활성)

  • Nam, Mi-Hyun;Lee, Hyun-Sun;Hong, Chung-Oui;Koo, Yoon-Chang;Seo, Mun-Young;Lee, Kwang-Won
    • Korean Journal of Food Science and Technology
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    • v.42 no.2
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    • pp.210-216
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    • 2010
  • Rosa rugosa has traditionally been used as a folk remedy for diabetes. The objective of this study was therefore to demonstrate the inhibition of endothelial dysfunction activities through antioxidants and the anti-glycation of Rosa rugosa roots. Dried roots of Rosa rugosa were boiled in methanol for three hours, evaporated and lyophilized with a freeze-dryer. The methanolic extract of Rosa rugosa roots (RRE) was tested for antioxidant activities by measuring total polyphenol (TP) content, flavonoid content, 1,1-diphenyl-2-picrylhydrazyl free radical-scavenging activity (DPPH) assay, and ferric-reducing antioxidant power (FRAP) assay. The total TP content, flavonoid content, FRAP value, and $DPPHSC_{50}$ are $345.2\;{\mu}g$ gallic acid equivalents/mg dry matter (DM), $128.1\;{\mu}g$ quercetin equivalents/mg DM, 2.2 mM $FeSO_4$/mg DM and $34.2\;{\mu}g$ DM/mL, respectively. Treatment of RRE significantly lowered fluorescent formation due to advanced glycation reaction. In addition, reactive oxygen species (ROS) scavenging assay, monocyte adherent assay and transendothelial electrical resistance (TEER) assay were performed to investigate the possibility that RRE improves endothelial dysfunction-induced diabetic complications. The adhesion of THP-1 to treated HUVEC with RRE ($100\;{\mu}g/mL$; 33% and $500\;{\mu}g/mL$; 75%) was significantly reduced compared to HUVEC stimulated by glyceraldehydes-AGEs (advanced glycation end product). The TEER value ($88\;{\Omega}{\cdot}cm^2$) of stimulated HUVEC by glyceraldehydes-AGEs was reduced compared to non-stimulation ($113\;{\Omega}{\cdot}cm^2$). However, normalization with RRE increased endothelial permeability in a dose-dependent manner ($100\;{\mu}g/mL$; $102\;{\Omega}{\cdot}cm^2$ and $500\;{\mu}g/mL$; $106\;{\Omega}{\cdot}cm^2$). Thus, these results suggest that Rosa rugosa roots could be a novel candidate for the prevention of diabetic complications through antioxidants and inhibition of advanced glycation end product formation.

Analysis of total polyphenol content and antioxidant activity in puffed oats (팽화 귀리에서의 총 폴리페놀 함량 변화 및 항산화능 비교)

  • Lee, Ji Hae;Son, Yurim;Lee, Byoung-Kyu;Lee, Byongwon;Kim, Hyun-Joo;Park, Ji-Young;Lee, Hyun Seok;Kim, Jin Suk;Park, Hyoung-Ho;Han, Ouk-Kyu;Han, Sangik;Lee, Yu Young
    • Korean Journal of Food Science and Technology
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    • v.50 no.1
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    • pp.117-121
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    • 2018
  • Puffing process modifies the chemical and physical properties of the grains. In this study, oats were puffed by subjecting them to pressure of 1.0 and 1.2 MPa, following which the bioactive constituents and antioxidant activities in the oat extracts were investigated. The polyphenol content in puffed oat extracts increased in a pressure-dependent manner (109 and 157 mg gallic acid equivalent/100 g at 1.0 and 1.2 MPa, respectively). In addition, gallic acid was synthesized after puffing ($518{\mu}g/g$ of extract at 1.0 MPa) and was the most abundant phenolic acid in puffed oats. The antioxidant activities, which were determined by 1, 1-diphenyl-2-picrylhydrazyl (DPPH) and 2, 2-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid (ABTS) radical scavenging activities, were improved in oat extracts after puffing (+245 and +184% at 1.2 MPa, respectively). In conclusion, puffing process of oats increased the extractability of polyphenols, including gallic acid, which positively affected its antioxidant activities. These results will provide useful information when using puffed oats for food production.