• Title/Summary/Keyword: 분리 및 선별

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Mechanical evaluation of SiC-graphite interface of seed crystal module for growing SiC single crystals (탄화규소 단결정 성장을 위한 종자결정모듈의 탄화규소-흑연 간 접합계면의 기계적 특성 평가)

  • Kang, June-Hyuk;Kim, Yong-Hyeon;Shin, Yun-Ji;Bae, Si-Young;Jang, Yeon-Suk;Lee, Won-Jae;Jeong, Seong-Min
    • Journal of the Korean Crystal Growth and Crystal Technology
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    • v.32 no.5
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    • pp.212-217
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    • 2022
  • Large thermal stress due to the difference between silicon carbide and graphite's coefficients of thermal expansion could be formed during crystal growing process of silicon carbide (SiC) at high temperature. The large thermal stress could separate the SiC seed crystals from graphite components, which bring about the drop of the seed crystal during crystal growth. However, the bonding properties of SiC seed crystal module has hardly reported so far. In this study, SiC and graphite were bonded using 3 types of bonding agents and a three-point bending tests using a mixed-mode flexure test were conducted for the bonded samples to evaluate the bonding characteristics between SiC and graphite. Raman spectroscopy, X-ray Photoelectron Spectroscopy, and X-ray Computed Tomography were used to analyze the bonding characteristics and the microstructures of the SiC-graphite interfaces bonded with the bonding agents. As results, an excellent bonding agent was chosen to fabricate SiC seed crystal module with 50 mm in diameter. An SiC single crystal with 50 mm in diameter was successfully grown without falling out during top seeded solution growth of SiC at high temperature.

Production and Characterization of the Beneficial β-glucuronidase Inhibitor from non pathogenic wild yeast, Candida oleophila WP5-19-1, and its effects on gut microbes (비병원성 야생효모 Candida oleophila WP5-19-1로부터 장내유용세균 유해 효소 β-glucuronidase 저해제의 생산 및 특성)

  • Seon-Jeong Park;Jong-Soo Lee
    • The Korean Journal of Mycology
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    • v.50 no.1
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    • pp.31-40
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    • 2022
  • We aimed to produce a potent β-glucuronidase inhibitor from wild yeast that could inactivate toxic substances in the colon. Culture supernatants and cell-free extracts of non-pathogenic wild yeasts were prepared and their β-glucuronidase inhibitory activities were measured. Cell-free extract from Candida oleophila WP5-19-1 showed the highest β-glucuronidase inhibitory activity (49.0%). The β-glucuronidase inhibitor was maximally produced (IC50 value; 8.4 mg) when C. oleophila WP5-19-1 was cultured in potato dextrose medium containing 5% dextrose (initial pH; 6.0) at 30℃ for 24 hours. β-glucuronidase inhibitor of C. oleophila WP5-19-1 was partially purified by trypsin hydrolysis, ultrafiltration (3 kDa), and Sephadex G-50 filtration. The partially purified β-glucuronidase inhibitor was stable from 30℃ to 60℃ and at pH 6.0 9.0, and showed residual inhibitory activity of about 80%.

Physiological Functionalities and Enzyme Activities of Non-pathogenic Pigmented Wild Yeasts (비병원성 야생 색소 효모들의 생리활성과 효소활성)

  • Jeong-Su Moon;Seung-A Shin;Seung-Eun Baek;Jong-Soo Lee
    • The Korean Journal of Mycology
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    • v.51 no.3
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    • pp.205-217
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    • 2023
  • This study aimed to produce novel bioactive compounds from non-pathogenic pigmented wild yeasts. Culture supernatants and cell-free extracts of non-pathogenic pigmented yeast strains were prepared, and their physiological functionalities and enzyme activities were measured. Cell-free extracts from Rhodosporidium paludigenum HHGG35-1 and culture supernatants from Rhodosporidium diobovatum NMD18-1 demonstrated very high antioxidant activity (76.6%) and anti-gout xanthin oxidase inhibitory activity (86.2%), respectively. Maximal production of the antioxidants (76.9%) was obtained when Rh. Paludigenum HHGG35-1 was cultured in a yeasts extract-peptone-dextrose (YPD) medium (pH 6.5) at 30℃ for 24 h. The xanthin oxidase inhibitor was also maximally produced (91.6%) when Rh. Diobovatum NMD18-1 was cultured at 30℃ for 96h in a YPD medium (pH 6.5). Rh. Paludigenum HHGG35-1 was oval in shape and formed ascospre. The Rh.diobovatum NMD18-1 specimen displayed dimensions of 1.6 × 1.6 ㎛ and produced ascospores; however, it did not form pseudomycelium. Both of Rh. Paludigenum HHGG35-1 and Rh. Diobovatum NMD18-1 grew well in a 40%-glucose-containing YPD medium and 10%-NaCl-containing YPD medium.

Identification and Functional Analysis of Escherichia coli RNase E Mutants (Escherichia coli 리보핵산 내부분해효소 RNase E의 돌연변이체 선별 및 특성분석)

  • Shin, Eun-Kyoung;Go, Ha-Young;Kim, Young-Min;Ju, Se-Jin;Lee, Kang-Seok
    • Korean Journal of Microbiology
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    • v.43 no.4
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    • pp.325-330
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    • 2007
  • RNase E is an essential Escherichia coli endoribonuclease that plays a major role in the decay and processing of a large fraction of RNAs in the cell and expression of N-terminal domain consisted of 1-498 amino acids (N-Rne) is sufficient to support normal cellular growth. By utilizing these properties of RNase E, we developed a genetic system to screen for amino acid substitutions in the catalytic domain of the protein (N-Rne) that lead to various phenotypes. Using this system, we identified three kinds of mutants. A mutant N-Rne containing amino acid substitution in the S1 domain (I6T) of the protein was not able to support survival of E. coli cells, and another mutant N-Rne with amino acid substitution at the position 488 (R488C) in the small domain enabled N-Rne to have an elevated ribonucleolytic activity, while amino acid substitution in the DNase I domain (N305D) only enabled N-Rne to support survival of E. roli cells when the mutant N-Rne was over-expressed. Analysis of copy number of ColEl-type plasmid revealed that effects of amino acid substitution on the ability of N-Rne to support cellular growth stemmed from their differential effects on the ribonucleolytic activity of N-Rne in the cell. These results imply that the genetic system developed in this study can be used to isolate mutant RNase E with various phenotypes, which would help to unveil a functional role of each subdomain of the protein in the regulation of RNA stability in E. coli.

Electrofusion of Yeast Cells and Their Genetic Analysis Using RAPD-PCR (효모세포의 전기융합 및 융합세포의 RAPD-PCR을 이용한 유전적 분석)

  • Kim, Seung;Kim, Jae-Sung;Sapkota, Kumar;Park, In-Sung;Cho, Moon-Gu;Park, Yeal;Chun, Hong-Sung;Choi, Bong-Suk;Park, Se-Eun;Choi, Han-Suk;Kim, Myung-Kon;Kim, Sung-Jun
    • Applied Biological Chemistry
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    • v.49 no.3
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    • pp.186-191
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    • 2006
  • In the present study, an attempt has been made to produce hybrid yeast strains of different useful and dominant characteristics. The hybrid yeast strains were produced by electrofusion and their genetic analysis were performed by RAPD-PCR (random amplified polymorphic DNA-polymerase chain reaction). The protoplast of Saccharomyces cerevisiae KCTC 7904 and Zygosaccharomyces rouxii KCTC 7966 were obtained above 92% when treated with lyticase at $30^{\circ}C$ for $60{\sim}90$ min after the pretreatment of $1{\sim}2%$ 2-mercaptoethanol at $30^{\circ}C$ for $15{\sim}20$ min. The fusant was produced from paired protoplast stage under the electric pulse at high frequency conditions (1.5 MHz/50 pV, 615 $V/256\;{\mu}sec$) within glass-platinum made electrofusion chamber. Changes in RAPD patterns in mother cells and hybrid cells proved that the fusant contains two types of yeast gene originated from its parent. Furthermore, fermentation characters exhibits by the fusant cell confirmed its genetic changes. These results suggest that genetically stable hybrid yeast strains of economic importance can be produced by electrofusion technique and these electrofused yeast cells have an enormous impact in biotechnology and biomedicine.

Isolation and Identiffication of Acinetobacter koreensis sp. nov. from Jang-Baek Waterfall (장백 폭포에서 분리한 Acinetobacter koreensis sp. nov.의 보고)

  • Lee, Ha-Yan;Yoo, Yong-Kyu;Seo, Pil-Soo;Lee, Jung-Sook;Lee, Keun-Chul;Lee, Sang-Seob
    • Korean Journal of Microbiology
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    • v.43 no.1
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    • pp.66-71
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    • 2007
  • Two isolates of genus Acinetobacter were obtained from Jang-Baek waterfall in North Korea. Morphological characteristics of the isolated 2 strains were Gram-negative, aerobic and rod shape bacteria. Physiological and biochemical characterization of the isolated 2 strains were some different aspect from those of type strains. 16S rDNA sequence analysis showed that the two isolates shared 99.9% sequence similarity. Strains JB10 and $JB15^{T}$ were shown to belong to the Gammaproteobacteria and showed the highest levels of sequence similarity to Acinetobacter tandoii $4N13^{T}$ (97.3%), Acinetobacter haemolyticus $ATCC17906^{T}$ (97.2%), Acinetobacter johnsonii $DSM6963^{T}$ (97.2%), Acinetobacter junii $DSM6964^{T}$ (96.7%), Acinetobacter schindleri $LUH5832^{T}$ (97.0%) and Acinetobacter ursingii $LUH3792^{T}$ (96.6%). The major cellular fatty acid in Acinetobacter type strains and isolated strains included $C_{18:1}\;{\omega}9c\;and\;C_{16:1}\;{\omega}7c/C_{15:0}\;iso\;2OH$. Eventhough it was ascertained that the isolated strains were closely related to genus Acinetobacter, physiological and biochemical characteristics and the result of the isolated strains 16S rDNA analysis indicate some different aspects from those of type strains of genus Acinetohacter It is considered that the isolated JB10 (=KEMC 52-093) and $JB15^{T}\;(=KEMC\;52-094^{T})$ strains be new species of genus Acinetobacter. We name it as Acinetobacter koreensis sp. nov.

DNA damages with Fpg/Endo Ⅲ FLARE Assay in cynomolgus monkeys exposed to stainless steel welding fume (용접흄 흡입노출 영장류에서 Fpg/Endo Ⅲ FLARE Assay를 이용한 DNA 손상 및 회복)

  • Rim, Kyung Taek;Kim, Soo Jin;Chung, Yong Hyun;Kim, Hyeon Yeong;Maeng, Seung Hee;Yu, Il Je
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.17 no.4
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    • pp.272-281
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    • 2007
  • 선박제조업을 비롯한 운송업 및 건축업 등의 다양한 분야에서 용접기술이 이용되어 옴에 따라 용접근로자들에 대한 산업보건학적 관심이 높아지고 있다. 노출정도가 다양하기는 하지만 용접흄은 6가 크롬을 비롯한 금속화합물과 유해가스, 화학물질 등을 복합적으로 포함하고 있는 스테인레스 스틸 용접흄에 대한 유전독성영향을 평가하기 위하여 흡입챔버를 이용, 실험동물인 영장류에 스테인레스 스틸 용접흄을 노출시키고 혈액 내 lymphocytes에 생성된 용접흄 노출농도 및 시간별 DNA 손상정도 및 그 회복효소를 측정함으로써, 유해성이 완전하게 확인되지 않은 용접흄에 노출되어 나타날 수 있는 암을 비롯한 심각한 건강영향을 예방하기 위한 각 지표들을 찾아 그 유용성을 비교하고자 하였다. 영장류를 노출시키기 위해 robotic arm을 장치한 영장류 흡입노출 시스템을 개발하였으며, 이 노출 시스템을 이용하여 수컷 영장류 6마리에 대해 용접흄 노출시험을 실시하였는데 실험군은 대조군 2, 저농도 ($31mg/m^3$) 노출군 2, 고농도 ($63mg/m^3$) 노출군 2마리로 구성하였고, 1일 2시간씩 일주일에 5일 동안 용접흄에 노출시켰다. 노출 농도는 지속적으로 모니터링 하였고, 노출과정 중에 영장류의 혈액을 채취하여 lymphocytes를 분리, 단세포 DNA 손상을 선별하기 위해 DNA 손상회복 효소인 E. coli formamidopyrimidine-DNA glycosylase (Fpg)와 endonuclease Ⅲ (Thymine Glycol-DNA glycosylase) 투여와 Comet asaay (single cell gel electrophoresis, 단세포겔전기영동기법)를 결합시켜 이용하는 Fpg/Endo III FLARE 분석법을 사용하였다. Fpg enzyme에 의한 olive tail moment값의 변화는 16주 노출군부터 노출부검(34주)군 까지 노출농도가 높아짐에 따른 olive tail moment 기하평균 값의 양 반응관계를 보기는 어렵지만, 고농도군의 경우 27주 노출군에서 가장 높은 olive tail moment 값을 보이고 이후 차츰 감소하였다. 한편 16주에서 22주까지의 노출기간에서는 대조군에 비해 노출군에서 DNA손상정도(olive tail moment값)는 모두 유의하게 높았으나, 6, 12, 18, 25, 31, 33, 35주간 노출하였을 때는 다른 결과를 보였다. 각 실험군의 Fpg enzyme에 의한 tail length값의 분포를 살펴볼 때, 저농도군 및 고농도군에서 27주간 노출하였을 때 가장 높은 tail length 값을 보이고 이후 차츰 감소하는 경향을 보였다. 또한 16, 22주간 노출하였을 때 대조군에 비해 노출군에서 tail length 값이 유의하게 높았으나, 20주간에서만 양 반응관계가 관찰되었고, 다른 주간에서는 양 반응 및 기간 반응관계를 나타내지는 않았다. Endo III enzyme에 의한 olive tail moment값의 변화는 기간별 노출군에서 대조군에 비해 높은 DNA손상정도(olive tail moment값)를 나타내는 결과들이 있었지만, 10, 12, 16, 22, 25, 31주간 노출하였을 때 등 상당수 노출기간에서 반응관계를 나타내지는 않았다. 각 실험군의 Endo III enzyme에 의한 tail length값의 분포를 살펴볼 때, 18, 20, 27, 33주간 노출하였을 때 대조군에 비해 노출군에서 tail length 값이 조금 높았지만, 양 반응 및 기간 반응관계를 보이지 않았고 수치의 크기가 불규칙하게 변화하였다. 즉, DNA에 있어 산화된 pyrimidine을 형성하여 손상된 부위의 염기를 제거함으로써 AP site (abasic site)를 만들고 이들이 Comet assay를 통해 break로 전환된 것을 포함한 DNA손상을 측정하기 위하여 endonuclease III (Endo III)를 첨가시킨 Endo III FLARE 분석법을 실시한 결과, 본 연구에서 나타난 결과는 용접흄 노출 영장류에서 Olive tail moment 및 tail length 공히 노출량 및 노출기간 반응관계를 볼 수 없었다. Endo III FLARE 분석법을 통한 산화적 DNA 손상지표는 영장류에 적용하기에는 적응반응현상으로 대조군과 유의한 차이도 관찰할 수 없었고 더욱이 역으로 대조군에서의 자연발생적 수치가 더 높아질 수 있어 용접흄 노출 영장류의 모니터링 지표로 사용하기에는 제한점이 있었다.

Immunogenicity and Reactogenicity of Inactivated HM175 Strain Hepatitis A Vaccine in Healthy Korean Children (건강한 한국 소아에서 HM175주 A형 간염 불활화 백신의 면역원성 및 이상반응에 관한 연구)

  • Kim, Chang Hwi;Pyun, Bok Yang;Hong, Young Jin;Kang, Jin Han
    • Pediatric Infection and Vaccine
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    • v.7 no.1
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    • pp.120-128
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    • 2000
  • Purpose : Active immunization against hepatitis A with an inactivated vaccine reveals excellent immunogenicity, tolerability and protective efficacy. Inactivated hepatitis A vaccines have been selectively used since 1996 in Korea to prevent hepatitis A. This study was performed to assess the immunogenicity and reactogenicity after two doses of HM175 strain hepatitis A vaccine in healthy Korean children. Methods : 128 healthy children(M/F; 65/63) aged 1 to 15 years, who were seronegative for hepaitatis A, participated in this study. A alum-adsorbed vaccine containing 720 EL.U of antigen form HM175 hepatitis A strain per 0.5 mL dose was injected intramuscularly on the deltoid area. The second dose was given 6 months later, Anti-HAV antibodies were measured by ELISA before and 1 month after each vaccination to assess the immunogenicity. Any local and general adverse events were reported by patients parents with the prepared questionnaire after each vaccination. Results : 120 volunteers(M/F; 60/60) completed the whole series of the study. Seroconversion occurred in all cases after primary and booster vaccination. The mean anti-HAV antibody titer after primary vaccination was 389.2mIU/mL, and 3,609mIU/mL after booster vaccination. And levels of anti-HAV antibodies after booster immunization were significantly higher in female children. The most common local adverse event was soreness on the injection site, but it was mild and resolves within 3 days. Fever was not reported after booster vaccination. Conclusion : Based on these data, we conclude that the inactivated HM175 strain hepatitis A vaccine is highly immunogenic and tolerable in healthy Korean children.

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Survival Ability of Porcine Oocytes Frozen-Thawed by Open Pulled Straw Method (Open Pulled Straw 방법에 의해 동결-융해된 돼지난자의 생존능력)

  • 김세웅;박춘근;정희태;양부근;김정익
    • Journal of Embryo Transfer
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    • v.16 no.2
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    • pp.117-125
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    • 2001
  • Vitrification of oocytes has been applied recently fur pigs, but remains elusive. The purpose of this study is to investigate the effects of vitrification in open pulled straws (OPS) on in vitro survival of porcine oocytes. When immature follicular oocytes frozen-thawed were cultured for in vitro maturation, maturation rates to metaphase-II stage were higher in oocytes with (25%) than without (15%) cumulus cells. After In vitro fertilization of oocytes frozen-thawed, the maturation rates were also significantly (P<0.05) higher in oocytes with (41%) that than without (17%) cumulus cells. However, the penetration rates were higher in oocytes without (19%) that than with (9%) cumulus. In another experiment, porcine oocytes matured in vitro were frozen and thawed for in vitro fertilization. The penetration rates were higher than in oocytes without (35%) that than with (26%) cumulus cells. However, the proportions of oocytes dead after in vitro fertilization were significantly (P<0.05) higher in oocytes with that than without cumulus cells. On the other hand, the rates of penetration and dead oocytes at 6 h after in vitro fertilization were not significant differences between oocytes with and without cumulus cells. However, the proportions of dead oocytes with (18%) and without (16%) cumulus cells were higher than in oocytes of control group (0%). These finding indicated the possible broader application for OPS, as they demonstrated that the maturation and fertilization in vitro by frozen-thawed oocytes may be accompained by cumulus cells and culture periods according to the requirements of the survival ability after freezing of mature and immature oocytes in pigs.

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Relationships on Magnitude and Frequency of Freshwater Discharge and Rainfall in the Altered Yeongsan Estuary (영산강 하구의 방류와 강우의 규모 및 빈도 상관성 분석)

  • Rhew, Ho-Sang;Lee, Guan-Hong
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.16 no.4
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    • pp.223-237
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    • 2011
  • The intermittent freshwater discharge has an critical influence upon the biophysical environments and the ecosystems of the Yeongsan Estuary where the estuary dam altered the continuous mixing of saltwater and freshwater. Though freshwater discharge is controlled by human, the extreme events are mainly driven by the heavy rainfall in the river basin, and provide various impacts, depending on its magnitude and frequency. This research aims to evaluate the magnitude and frequency of extreme freshwater discharges, and to establish the magnitude-frequency relationships between basin-wide rainfall and freshwater inflow. Daily discharge and daily basin-averaged rainfall from Jan 1, 1997 to Aug 31, 2010 were used to determine the relations between discharge and rainfall. Consecutive daily discharges were grouped into independent events using well-defined event-separation algorithm. Partial duration series were extracted to obtain the proper probability distribution function for extreme discharges and corresponding rainfall events. Extreme discharge events over the threshold 133,656,000 $m^3$ count up to 46 for 13.7y years, following the Weibull distribution with k=1.4. The 3-day accumulated rain-falls which occurred one day before peak discharges (1day-before-3day -sum rainfall), are determined as a control variable for discharge, because their magnitude is best correlated with that of the extreme discharge events. The minimum value of the corresponding 1day-before-3day-sum rainfall, 50.98mm is initially set to a threshold for the selection of discharge-inducing rainfall cases. The number of 1day-before-3day-sum rainfall groups after selection, however, exceeds that of the extreme discharge events. The canonical discriminant analysis indicates that water level over target level (-1.35 m EL.) can be useful to divide the 1day-before-3day-sum rainfall groups into discharge-induced and non-discharge ones. It also shows that the newly-set threshold, 104mm, can just separate these two cases without errors. The magnitude-frequency relationships between rainfall and discharge are established with the newly-selected lday-before-3day-sum rainfalls: $D=1.111{\times}10^8+1.677{\times}10^6{\overline{r_{3day}}$, (${\overline{r_{3day}}{\geqq}104$, $R^2=0.459$), $T_d=1.326T^{0.683}_{r3}$, $T_d=0.117{\exp}[0.0155{\overline{r_{3day}}]$, where D is the quantity of discharge, ${\overline{r_{3day}}$ the 1day-before-3day-sum rainfall, $T_{r3}$ and $T_d$, are respectively return periods of 1day-before-3day-sum rainfall and freshwater discharge. These relations provide the framework to evaluate the effect of freshwater discharge on estuarine flow structure, water quality, responses of ecosystems from the perspective of magnitude and frequency.