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Characterization of Grain Amino Acid Composition and Proteome Profile of a High-lysine Barley Mutant Line M98 (고-Lysine 보리 돌연변이 계통 M98 종실의 아미노산 조성 및 Proteome Profile 특성)

  • Kim, Dea-Wook;Kim, Hong-Sik;Park, Hyoung-Ho;Hwang, Jong-Jin;Kim, Sun-Lim;Lee, Jae-Eun;Jung, Gun-Ho;Hwang, Tae-Young;Kim, Jung-Tae;Kim, Si-Ju;Rakwal, Randeep;Kwon, Young-Up
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.57 no.2
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    • pp.171-181
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    • 2012
  • Lysine is the first limiting essential amino acid in cereals for humans and monogastric animals, although its content is generally low. A chemically induced high-lysine barley mutant, M98, has an agronomically undesirable shrunken endosperm trait. In order to obtain detailed insight into the atypical traits of M98 grains, we characterized amino acid composition and protein profiles of M98 and its parent cultivar Chalssalbori. Among a total of 16 amino acids, the percentage of each of the 7 amino acids, including lysine, was 1.2~1.8 times higher in M98, comparing to Chalssalbori. The percentage of proline and its precursor, glutamic acid, in M98 was about the half of that of the amino acids in Chalssalbori, but arginine synthesized from glutamic acid was 1.8 times higher in M98, compared that in the parent cultivar. Theses results indicated that the mutation in M98 grains might alter the proportion of amino acids linked to each other in a biosynthetic pathway. A comparison of grain proteome profiles between Chalssalbori and M98 revealed 70 differentially expressed protein spots, where 45 protein spots were up-regulated and 25 protein spots down-regulated in M98 compared to those in Chalssalbori. Of these changed protein spots, 53 were identified using nano-electrospray ionization liquid chromatography mass spectrometry. Most of these identified proteins were involved in various biological processes. In particular, 28 protein spots such as ${\beta}$-amylase, serpins and B3-hordein were identified as proteins associated with the atypical traits of M98. It was thought that a genetic study on the unique protein profile of M98 would be needed to develop an agronomically feasible barley cultivar with high-lysine trait.

Relation between Health Examination Outcome and Intake of Soy Food and Isoflavone among Adult Male in Seoul (서울 거주 성인 남자의 대두식품 및 이소플라본 섭취와 각종 건강지표와의 관련성 분석)

  • Lee, Min-June;Sohn, Chun-Young;Kim, Ji-Hyang
    • Journal of Nutrition and Health
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    • v.41 no.3
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    • pp.254-263
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    • 2008
  • This study was conducted to analyze the effect of isoflavone intake on prevention of chronic disease in middle and old aged man. In this study we used FFQ (Food frequency questionnaire) and the isoflavone intake level of the subjects was 25.10 mg per day. We divided the subjects into three group -high, medium, low isoflavone intake level- and investigated the relation among isoflavone intake level and clinical/anthropometric characteristics. The intake of isoflavone was inversely related with the body fat in male subjects. And we also divided the subjects into 2 groups with normal and abnormal clinical/anthropometric risk factor. The isoflavone intake level of the abnormal group with high TG, high WHR and high body fat was lower than the normal group. The main food source of isoflavone was soybean curd, bean sprout, soybean paste, soybean and soy milk, and we also investigated the relation between frequency of soybean food and anthropometric and clinical variables. The frequencies of soybean curd, soybean paste, soybean broth, soy milk, bean sprouts, peanuts, soybean and dambuk as well as intake of isoflavone were inversely correlated with some anthropometric and clinical variables such as blood pressure, TG, BMI, % body fat, and waist-hip ratio, whereas positively correlated with HDL cholesterol, muscle mass and bone density. We suggest that high consumption of soy products and isoflavone is associated with decreased blood lipid and body fat in middle and old aged man and might be useful for prevention cardiovascular diseases. From this study, we obtained valuable basic information on recommended isoflavone intake level and guidelines for the prevention of some chronic diseases/health problems.

Growth promoting effect of combined gonadotropin releasing hormone analogue and growth hormone therapy in early pubertal girls with predicted low adult heights (예측성인신장이 작은 조기사춘기 여아에서 성선자극호르몬 방출호르몬 효능약제와 성장호르몬 병합치료의 성장획득 효과)

  • Hong, Eun-Jeong;Han, Heon-Seok
    • Clinical and Experimental Pediatrics
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    • v.50 no.7
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    • pp.678-685
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    • 2007
  • Purpose : Recent reports pointed out that gonadotropin releasing hormone analogue (GnRHa) therapy alone is not so promising for improving adult height in precocious puberty. So, that we studied the growth promoting effect of combined therapy with GnRHa and growth hormone (GH) in early pubertal girls. Methods : Twenty three early pubertal girls ($9.73{\pm}1.59yr$) with predicted adult heights (PAH) below-2 standard deviation score (SDS) were included. They were divided into two groups as follows; Group I before menarche (n=19) and Group II after menarche (n=4). After combined therapy, various growth parameters were compared between two groups and between the before and after therapy. Results : Between the two groups before therapy, chronologic age (CA), growth velocity (GV), body mass index (BMI), target height (TH), PAH and serum insulin-like growth factor binding protein-3 were not different, but BA, height and difference between bone age (BA) and CA were significantly higher and insulin-like growth factor-1 (IGF-1) was marginally higher in group II. After therapy, BA still remained higher in group II, but other parameters were not different. In both groups, after therapy, the difference between BA and CA, the ratio of BA over CA, and GV were significantly decreased, but PAH, height SDS and BMI were significantly increased. Regarding IGF-1 level, a significant increase was noted in group I, but not in group II. Conclusion : With combined therapy of GnRHa and GH, PAH in early pubertal girls might be improved significantly and even approach TH. Among them, those who were before menarche might have greater potential for the height gain than those after menarche in view of IGF-1 changes during therapy.

Changes of Protein Profiles in Cheonggukjang during the Fermentation Period (전통 청국장의 발효 기간 동안 변화하는 수용성 단백질 개요)

  • Santos, Ilyn;Sohn, Il-Young;Choi, Hyun-Soo;Park, Sun-Min;Ryu, Sung-Hee;Kwon, Dae-Young;Park, Cheon-Seok;Kim, Jeong-Hwan;Kim, Jong-Sang;Lim, Jin-Kyu
    • Korean Journal of Food Science and Technology
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    • v.39 no.4
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    • pp.438-446
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    • 2007
  • The fermented soybean product, cheonggukjang, is favored by many people, partly due to its bio-functional ingredients. Since the fermentation process of cheonggukjang is mediated by enzymes, including proteases, produced by microbes, analysis of the proteome profile changes in cheonggukjang during fermentation would provide us with valuable information for fermentation optimization, as well as a better understanding of the formation mechanisms of the bio-functional substances. The soluble proteins from cheonggukjang were prepared by a phenol/chloroform extraction method, in order to remove interfering molecules for high resolution 2-D gel analysis. Proteomic analysis of the cheonggukjang different fermentation periods suggested that most of the soluble soy proteins were degraded into smaller forms within 20hr, and many microbial proteins, such as mucilage proteins, dominated the soluble protein fraction. The proteomic profile of cheonggukjang was very different from natto, in terms of the 2-D gel protein profile. Among the separated protein spots on the 2-D gels, 50 proteins from each gel were analyzed by MALDI-TOF MS and PMF for protein identification. Due to database limitations with regard to soy proteins and microbial proteins, identification of the changed proteins during fermentation was restricted to 9 proteins for cheonggukjang and 15 for natto. From de novo sequencing of the proteins by a tandem MS/MS, as well as by database searches using BLASTP, a limited number of proteins were identified with low reliability. However, the 2-D gel analysis of proteins, including protein preparation methods, remains a valuable tool to analyze complex mixtures of proteins entirely. Also, for intensive mass spectrometric analysis, it is also advisable to focus on a few of the interestingly changed proteins in cheonggukjang.

Inhibitory Effect of Scopoletin Isolated from Sorbus commixta on TNF-α-Induced Inflammation in Human Vascular Endothelial EA.hy926 Cells through NF-κB Signaling Pathway Suppression (마가목 수피에서 분리한 scopoletin의 EA.hy926 혈관내피세포에서 NF-κB 신호전달을 통한 TNF-α로 유도된 혈관염증 저해 효과)

  • Kang, Hye Ryung;Kim, Hyo Jung;Kim, Bomi;Kim, Sun-Gun;So, Jai-Hyun;Cho, Soo Jeong;Kwon, Hyun Sook
    • Journal of Life Science
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    • v.30 no.4
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    • pp.343-351
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    • 2020
  • Sorbus commixta Hedl. has traditionally been used as a remedy for cough, asthma, and other bronchial disorders. In this study, three major triterpenoids-lupeol, β-sitosterol, and ursolic acid and a coumarin, scopoletin, were isolated from a CHCl3-soluble fragment of the bark of S. commixta. Their structures were identified by spectroscopic analyses, including mass spectrometry (MS), 1D-, and 2D- nuclear magnetic resonance spectroscopy (NMR), as well as by comparing the data with data reported in the literature. Scopoletin was isolated from this plant for the first time. It is a nutraceutical compound contained in many plants that has been reported to exert diverse biological activities, including anti-inflammatory effects. This study examined the inhibitory effect of scopoletin on TNF-α-induced vascular endothelial inflammation. Unlike the marginal impact of other compounds against low-density lipoprotein (LDL) oxidation and vascular endothelial inflammation, scopoletin showed remarkable activity on LDL oxidation (IC50 = 10.2 μM) and exerted vascular anti-inflammatory effects in EA.hy926 human endothelial cells activated by TNF-α. It suppressed the expression of adhesion molecules, such as ICAM-1, VCAM-1, and E-selectin, and blocked the adhesion between THP-1 monocytes and EA. hy926 endothelial cells. It also inhibited TNF-α-induced NF-κB translocation from the cytosol to the nucleus. Moreover, IκBα phosphorylation, which was increased by TNF-α treatment, was reduced after treatment with scopoletin. Thus, scopoletin inhibited TNF-α-induced vascular inflammation in endothelial cells by suppressing the NF-κB signaling pathway. These results demonstrate that owing to its anti-inflammatory activity in the vascular endothelium, scopoletin has the potential to inhibit atherosclerosis development.

Development of Mixed-bed Ion Exchange Resin Capsule for Water Quality Monitoring (수질 중 질소와 인 모니터링을 위한 혼합이온교환수지 캡슐의 개발)

  • Park, Chang-Jin;Kim, Dong-Kuk;Ok, Yong-Sik;Ryu, Kyung-Ryul;Lee, Ju-Young;Zhang, Yong-Seon;Yang, Jae-E
    • Applied Biological Chemistry
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    • v.47 no.3
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    • pp.344-350
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    • 2004
  • This study was conducted to develop and assess the applicability of mixed-bed ion exchange resin capsules for water quality monitoring in small agricultural watershed. Recoveries of resin capsules for inorganic N and P ranged from 96 to 102%. The net activation energies and pseudo-thermodynamic parameters, such as ${\Delta}G^{o\ddag},\;{\Delta}H^{o\ddag},\;and\;{\Delta}S^{o\ddag}$ for ion adsorption by resin capsules, exhibited relatively low values, indicating the process might be governed by chemical reactions such as diffusion. However, those values increased with temperature coinciding with the theory. The reaction reached pseudo-equilibrium within 24 hours for $NH_4-N\;and\;NO_3-N$, and only 8 hours for $PO_4-P$, respectively. The selectivity of resin capsules were in the order of $NO_3\;^-\;>\;NH_4\;^+\;>\;PO_4\;^{3-}$, coinciding with that of encapsulated Amberlite IRN-150 resin. At the initial state of equilibrium, the resin adsorption quantity was linearly proportional to the mass of ions in the streams, but the rate of movement leveled off, following Langmuir-type sorption isotherm. The overall results demonstrated that the resin capsule system was suitable for water quality monitoring in small agricultural watershed, judging from the reaction mechanism(s) of the resin capsule and the significance of model in field calibration.

Therapeutic Efficacy of Alendronate for Glucocorticoid Induced Metabolic Bone Disease in Children with Nephrotic Syndrome (신증후군 환아에서 스테로이드 유발 대사성 골질환에 대한 Alendronate의 치료 효과)

  • Lee Ji-Eun;Lee Hyun-Ok;Paik Kyung-Hoon;Lee Suk-Hyang;Jin Dong-Kyu
    • Childhood Kidney Diseases
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    • v.8 no.1
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    • pp.33-42
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    • 2004
  • Purpose : Children with nephrotic syndrome(NS) are under high risk for metabolic bone disease(MBD) as a complication of long-term glucocorticoid therapy. We prospectively evaluated the effect of oral bisphosphonate(alendronate) therapy in children with NS, which has proven efficacy in adult patients with glucocorticoid induced MBD. Methods : Among 58 children with NS, aged 5 to 8 years and haying a disease duration of more than 2 years, 30(51.7%) were enrolled to meet the selection criteria, less than -1.0 Z-scores of lumbar spine bone mineral density(BMD) by dual energy X-ray absorptiometry (DEXA). These 30 children were divided into three groups and each were assigned to receive alendronate, calcitriol, and no-medication, respectively for one year. Lumbar spine BMD was followed up every 6 months and the biochemical indexes were measured before and 1 year after the treatment. There were no significant difference among groups with respect to the average age, the initial BMD, and the cumulative steroid doses. Analysis of the treatment efficacy was done by the % change of BMD and by the changes in Z-scores of lumbar spine BMD. Results : Mean age and disease duration of patients at the initial lumbar spine BMD evaluation was $7.4{\pm}1.7$ years and $2.2{\pm}1.2$ years, respectively. Twenty-three of 30 children(76%) had osteopenia, and seven(23%) had osteoporosis. There was no difference in the biochemical values among the groups, before and 1 year after the treatment(P<0.05). Twenty two children(73.3%) with frequent relapsing or steroid dependant NS had more frequent MBD, compared to the 8 children(26.6%) with infrequent relapsing NS. The one year % changes of BMD were 8.56 in alendronate group, 5.79 in calcitriol group, and 1.9 in no-medication group. The changes in Z-score of lumbar spine BMD increased in the alendronate group and the calcitriol group, but not in the no-medication group. One year % changes of BMD were different among groups(P=0.0002). Significant differences were found between the alendronate and the no-medication group, and between the calcitriol and the no-medication group(P<0.05). There was no difference between the alendronate and the calcitriol group. No serious adverse effect was observed in the alendronate group. Conclusion : Children with NS receiving high dose steroids are under the high risk of BMD and should undergo regular BMD evaluation. Z-score of lumbar spine BMD was a useful parameter in diagnosing low bone mass in children. Alendronate weekly oral therapy was effective and relatively safe in increasing the lumbar spine BMD in children with NS having steroid induced MBD.

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Numerical analysis of FEBEX at Grimsel Test Site in Switzerland (스위스 Grimsel Test Site에서 수행된 FEBEX 현장시험에 대한 수치해석적 연구)

  • Lee, Changsoo;Lee, Jaewon;Kim, Geon-Young
    • Tunnel and Underground Space
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    • v.30 no.4
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    • pp.359-381
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    • 2020
  • Within the framework of DECOVALEX-2019 Task D, full-scale engineered barriers experiment (FEBEX) at Grimsel Test Site was numerically simulated to investigate an applicability of implemented Barcelona basic model (BBM) into TOUGH2-MP/FLAC3D simulator, which was developed for the prediction of the coupled thermo-hydro-mechanical behavior of bentonite buffer. And the calculated heater power, temperature, relative humidity, total stress, saturation, water content and dry density were compared with in situ data monitored in the various sections. In general, the calculated heater power and temperature provided a fairly good agreement with experimental observations, however, the difference between power of heater #1 and that of heater #2 could not captured in the numerical analysis. It is necessary to consider lamprophyre with low thermal conductivity around heater #1 and non-simplified installation progresses of bentonite blocks in the tunnel for better modeling results. The evolutions and distributions of relative humidity were well reproduced, but hydraulic model needs to be modified because the re-saturation process was relatively fast near the heaters. In case of stress evolutions due to the thermal and hydraulic expansions, the computed stress was in good agreement with the data. But, the stress is slightly higher than the measured in situ data at the early stage of the operation, because gap between rock mass and bentonite blocks have not been considered in the numerical simulations. The calculated distribution of saturation, water content, and dry density along the radial distance showed good agreement with the observations after the first and final dismantling. The calculated dry density near the center of the FEBEX tunnel and heaters were overestimated compared with the observations. As a result, the saturation and water content were underestimated with the measurements. Therefore, numerical model of permeability is needed to modify for the production of better numerical results. It will be possible to produce the better analysis results and more realistically predict the coupled THM behavior in the bentonite blocks by performing the additional studies and modifying the numerical model based on the results of this study.

한국인 좌심실 비대증 환자들에서 파브리병 선별검사의 의의

  • Park, Hyeong-Du;Jo, Seong-Yun;Lee, Su-Yeon;Jeon, Eun-Seok;Park, Seung-U;Lee, Sang-Hun;Lee, Sang-Cheol;Choe, Jin-O;Park, Seong-Ji;Jang, Seong-A;Kim, Hyeong-Gwan;Gi, Chang-Seok;Kim, Jong-Won;Jin, Dong-Gyu
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.14 no.2
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    • pp.135-141
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    • 2014
  • Objectives: Fabry disease (FD) is a lysosomal storage disease caused by the inappropriate accumulation of globotriaosylceramide (Gb3) in tissues due to a deficiency in the enzyme ${\alpha}$-galactosidase A. Hypertrophic cardiomyopathy is one of the chronic complications of FD. We tried to evaluate the prevalence of Fabry disease in the Korean patients with left ventricular hypertrophy (LVH). Methods: A total of 257 patients with LVH were recruited and they were 172 males (mean 56 years, range 30-81 years) and 84 females (mean 66 years, range 45-85 years). Urinary Gb3 was used to screen FD by high performance liquid chromatography-tandem mass spectrometry. Confirmatory tests were done by alpha-galactosidaseA activity using fluorometric assay and by GLA mutation analysis using sequencing. Results: Four patients were screening positive by urinary Gb3 analysis (cutoff, 25 ug/mmol creatinine). But, one female patient was diagnosed with FD confirmed by enzyme analysis in leukocytes as well as by genetic analysis (1/257 patients, 0.4%). She showed 54.3 ug/mmoL creatinine of Gb3 and 15.5 nmole/hr/mg protein (reference range, $55.2{\pm}12.7nmole/hr/mg$ protein) of alphagalactosidase A activity. And she had a heterozygous GLA mutation of c.796G>A (p.D266N). Her daughter was found to be a carrier for FD confirmed by GLA mutation analysis. Asymptomatic carrier showed 25.5ug/mmol creatinine of Gb3 and 42.5 nmole/hr/mg protein (reference range, $55.2{\pm}12.7nmole/hr/mg$ protein) of alpha-galactosidase A activity. Conclusions: The prevalence of FD in Koran patients with LVH was detected as 0.4%. Although the prevalence seems to be low, screening studies are of great importance for detecting hidden cases as well as for identifying other effected family members.

Conditions for Artificial Culture of Lemna Paucicostata and Potentiality as an Alternative Biomass Source (바이오매스 자원으로서의 Lemna Paucicostata의 인공배양조건과 이화학적 특성에 관한 연구)

  • Kwag, Jung-Hoon;Lee, Jin-Eui;Kim, Ki-Hye;Eum, Hye-Yeong;Shin, Jong-Suh;Ra, Chang-Six
    • Journal of Animal Environmental Science
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    • v.16 no.2
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    • pp.143-152
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    • 2010
  • Conditions for artificial culture of Lemna Paucicostata and its nutritional values were examined in this study. Lemna P. was cultured using artificial wastewater and a bioreactor (total volume $2,630\;cm^3$, working volume $2,240\;cm^3$) was operated at conditions of 6,250 lux and $28^{\circ}C$. Water flow affected the growth of Lemna P.: growth rate was very high (more than $1.1\;d^{-1}$) at a condition of no-water movement, but it was very low (less than $0.15\;d^{-1}$) when water moved slowly. The growth of Lemna P. was higher in $16h\;d^{-1}$ light cycle than in Sand $24h\;d^{-1}$, and it was also severely affected by the initial $NH_4$-N levels of wastewater. The growth rate of Lemna P. was high in lower $NH_4$-N level, indicating that the growth rate is in inverse proportion to $NH_4$-N concentration in wastewater. However, the contents of crude protein (CP) of Lemna P. were proportional to the initial $NH_4$-N concentration. The CP contents of Lemna P. cultured at 2, 10, 50 and 100 $NH_4$-N mg $L^{-1}$ was 18, 24, 37, 43%, respectively, showing the Lemna P. cultured at 50 and $100\;mg\;L^{-1}$ had similar protein contents to linseed (CP 35%), cottonseed (CP 38%) and soybean (CP 45%). Fat, protein, fiber, NDF and ADF contents of Lemna P. harvested at conditions of $16h\;d^{-1}$ light cycle and less than $2\;mg\;L^{-1}$ of $NH_4$-N level was 2.8, 18, 27, 20, 41 and 65.7%, respectively. Since the growth rate of Lemna P. was very high (more than $1.1\;d^{-1}$) at those conditions, it was convinced that mass production of valuable protein and fiber sources are feasible. In particular, since the Lemna P. has unsaturated fatty acids found mainly in animal fat as well as beneficial fatty acids to health such as C18:ln9c, C18:2n6c, C20:5n3 and C22:2, the Lemna P. biomass would be a highly valuable alternative feed source to grains.