• Journal of Sasang Constitutional Medicine
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• v.9 no.1
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• pp.315-337
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• 1997

In order to compare the dffects of $Galg{\breve{u}}nhaegit'ang$(葛根解肌湯) of "Dongeuisusebowon(東醫壽世保元)and Won's(元)-$Galg{\breve{u}}nhaegit'ang$(葛根解肌湯) of "Dongeuisasansinpyun(東醫四象新編)" on the immune respone, Sprague-Dawley male rats were used and randomly divided into four groups. Normal group was under normal condition, Control group was injected i.v. with 2mg/kg Methotrexate(MTX) on the 9th day and 11th day after sensitization with SRBC on the 5th day, $Galg{\breve{u}}nhaegit'ang$ group was fed with 1ml of $Galg{\breve{u}}nhaegit'ang$ and Won's-$Galg{\breve{u}}nhaegit'ang$ group was fed with 1ml of Won's-$Galg{\breve{u}}nhaegit'ang$ by oral during eighteen days. In the 9th day and the 11th day after oral feeding with medication, MTX was injected in tail of rats in order to reduce immune function. Leukocyte count, lymphocyte ratio, lymphocyte count of spleen, lymphocyte count of bone marrow, contact hypersensitivity to DNFB, morphologic change of thymus cell, and electropherogram of serum protein were estimated and compared according to the group. The results are as follows : 1. Before and after MTX injection, leukocyte(WBC) count was increased signigicantly in Won's-$Galg{\breve{u}}nhaegit'ang$ group compared to control group. $Galg{\breve{u}}nhaegit'ang$ group had no significant difference compared to control group. 2. Before and after MTX injection, lymphocyte ratio was not significantly different in Won's-$Galg{\breve{u}}nhaegit'ang$ group and in $Galg{\breve{u}}nhaegit'ang$ group compared to control group. 3. The lymphocyte count of spleen was increased significantly in $Galg{\breve{u}}nhaegit'ang$ group compared to control group and Won's-$Galg{\breve{u}}nhaegit'ang$ group. Won's-$Galg{\breve{u}}nhaegit'ang$ group had no significant difference compared to control group. 4. The lymphocyte count of bone marrow was increased significantly in Won's-$Galg{\breve{u}}nhaegit'ang$ group compared to control group and $Galg{\breve{u}}nhaegit'ang$ group. $Galg{\breve{u}}nhaegit'ang$ group had no significant difference compared to control group. 5. Contact hypersensitivity was increased significantly in Won's-$Galg{\breve{u}}nhaegit'ang$ group compared to other group. $Galg{\breve{u}}nhaegit'ang$ group had no significant difference compared to control groups. 6. In the morphologic change of thymus cell, control group compared to normal group had a indistinct boundary between cortex and medulla and lymphocyte cell density of thymus was low. $Galg{\breve{u}}nhaegit'ang$ group and Won's-$Galg{\breve{u}}nhaegit'ang$ group compared to control group had a definite boundary between cortex and medulla and lymphocyte cell density of thymus was high. 7. In the SDS-PAGE electropherogram of serum protein, Won's-$Galg{\breve{u}}nhaegit'ang$ group had a wide band of nearby 25,000 Dalton, and which meant IgG generated more actively. Considering this results, $Galg{\breve{u}}nhaegit'ang$ group and Won's-$Galg{\breve{u}}nhaegit'ang$ group have an effect on the depression of immune function induced by MTX, and especially Won's-$Galg{\breve{u}}nhaegit'ang$ group has an significant effect than $Galg{\breve{u}}nhaegit'ang$ group.

• KSBB Journal
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• v.13 no.2
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• pp.181-186
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• 1998

In order to obtain high-level production of recombinant human thrombopoietin (rhTPO) in insect cell line, HTI-TN-5B1-4 (TN5), conditions for optimal rhTPO expression such as multiplicity of infection (MOI), the cell density at infection, harvesting time and type of culture method as well as growth media were determined. When TN5 cells were cultured as anchorage-dependent state in 60-mm dish, cell density $2\times^6$ cells,MOI of 10 and Garvesting the culture media at 72 hr post-infection wrere the cinditions for highest rh TPO production. High production of rhTPO was also achieved by using EXPRESS FIVE serum free media rather than SF900II serum free media-1. Anchorage-dependent TN5 cells were adapted as a suspension culture when they were grown in the presence of heparin. TN5 cells were successfully cultured at 0.2 L scale in suspension culture without having aggregation. When TN5 cells were cultured as suspension state, cell density of $0.6\times10^6$ cells/mL, MOI of 1 and harvesting the culture media at 72 hr post-infection, gave the highest yield of rhTPO.

• Journal of Life Science
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• v.21 no.2
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• pp.309-315
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• 2011

Rice syrup meal (RSM) was enzymatically hydrolyzed using eight commercial proteases (Protamex, Neutrase, Flavourzyme, Alcalase, Protease M, Protease N, Protease A, Molsin F) for 4 hr at optimum pH and temperature. Proteolytic hydrolysates were examined in supernatant and precipitate using Lowry protein assay, semimicro Kjeldahl method and gravimetric method using weight difference before and after enzymatic hydrolysis. Although RSM contains a high amount of protein (71.2%), only a very small amount of protein was hydrolyzed. Two proteases (Protease M and Protease N) were found to be the most effective in the hydrolysis of RSM protein. In Lowry method, 57.5 and 59.0 mg protein/g RSM were hydrolyzed after Protease M and Protease N treatments, respectively. In gravimetric method, 80.0 and 85.4 mg protein/g RSM were hydrolyzed after Protease M and Protease N treatments. In Kjeldahl method, 67.43 and 70.43 mg protein/g RSM were hydrolyzed after Protamex and Protease N treatments, respectively. For synergistic effect, two or three effective commercial proteases (Protease M, Protease N and Protease A) were applied to RSM at one time. The highest hydrolysis of RSM protein was observed in both Lowry protein assay (80.3 mg protein/g RSM) and gravimetric methods (153.2 mg protein/g RSM) when three commercial proteases were applied at one time, suggesting the synergistic effect of those proteases.

• Korean Journal of Microbiology
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• v.40 no.4
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• pp.275-281
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• 2004

Our previous research has demonstrated that the bacterium, Pseudomonas sp. NFQ-l capable of utilizing quin­oline (2,3-benzopyridine) as the sole source of carbon, nitrogen, and energy was isolated and characterized [Yoon et ai. (2003) Kor. J. Biotechnol. Bioeng. 18(3):174-179]. In this study, we have found that Pseudomonas sp. NFQ-l could degrade quinoline as well as benzoate, and extended this work to characterize the catechol 1,2­dioxygenase (C1,2O) purified from the bacterium cultured in benzoate media. Initially, C1,2O has been purified by ammonium sulfate precipitation, gel permeation chromatography, and Source 15Q. After Source 15Q, puri­fication fold was increased to approximately 14.21 unit/mg. Molecular weight of C1,2O was about 33 kDa. Physicochemical characteristics (e.g., substrate specificity, Km, Vmax, pH, temperature and effect of inhibitors) of purified C1,2O were examined. C1,2O demonstrated the activity for catechol, 4-methylcatechol and 3-meth­ylcatechol as a substrate, respectively. The Km and Vmax value of C1,2O for catechol was 38.54 ${\mu}M$ and $25.10\;{\mu}mol{\cdot}min^{-1}{\cdot}mg^{-1}.$ The optimal temperature of C1,2O was $30^{\circ}C$ and the optimal pH was approximately 8.5. Metal ions such as $Ag^+,\;Hg^+,\;Ca^{2+},\;and\;Cu^{2+}$ show the inhibitory effect on the activity of C1,2O. N-terminal amino sequence of C1,2O was analyzed as ^1TVKISQSASIQKFFEEA^{17}.$ In this work, we found that the amino acid sequence of NFQ-l showed the sequence homology of 82, 71, 59 and $53\%$ compared with C1,2O from Pseudomonas aeruginosa PA0l, Pseudomonas arvilla C-1., P. putida KT2440 and Pseudomonas sp. CA10, respectively.

• Clinical and Experimental Pediatrics
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• v.50 no.2
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• pp.198-204
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• 2007

Purpose : The hope that arresting pubertal developement might increase final adult height has led to an attempt to use GnRH agonist (GnRHa) in children with early puberty and poor growth prognosis. We investigated the growth-promoting effect of GnRH agonists with or without growth hormone (GH) in girls with early puberty and decreased predicted adult height (PAH). Methods : Thirty five girls with advanced bone age and early pubertal signs were randomized for treatment for about 1 year with monthly GnRHa in group 1 (n=18), or with a combination of GH and GnRHa in group 2 (n=17). The following growth parameters were compared between groups, and the difference ($\Delta$) before and after treatment : chronological age (CA), bone age (BA), $\Delta$(BA-CA), height (HT), target height (TH), predicted adult height (PAH), $\Delta$ (TH-PAH), serum insulin-like growth factor (IGF-1) and insulin-like growth factor binding protein (IGFBP-3). Results : Before treatment, BA, TH, PAH Standard deviation scores (SDS), $\Delta$(TH-PAH) were not different between the two groups, but CA was higher in group 2 and $\Delta$(BA-CA) were higher in group 1 (P<0.05). After $1.06{\pm}0.93$ year of treatment, $\Delta$ (BA-CA) decreased and there were significant changes in PAH and $\Delta$ (TH-PAH), especially in group 2 (P<0.05 in group 1, and P<0.001 in group 2). In both groups, IGF-1 and IGFBP-3 were not different before and after treatment, but after treatment, IGF-1 level in group 2 was marginally higher than IGF-1 in group 1 (P<0.1). Conclusion : Compromised predicted adult height in girls with early puberty and advanced bone age was significantly improved with GnRH with/without GH treatment in the short-term period. The addition of GH to GnRHa results in a significant increase in PAH compared to GnRHa alone because GnRHa suppressed growth hormone-IGF-1 axis. For comparison of final adult height, further longitudinal follow-up will be needed.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.63 no.3
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• pp.229-238
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• 2018

The purpose of this study was to evaluate the effects of nitrogen fertilization amount and timing of application on protein content, dough properties, change in protein fraction, and bread loaf volume for Korean wheat cultivars, Baekkang, Joongmo2008, and Saekeumkang, for bread with a superior gluten composition during the grain filling stage. Protein content increased with an increase in the amount of N and timing of application. The SDS segmentation volume (SDSS) increased with an increase in N, but there was no effect of the timing of N application on SDSS. An increase in N amount and timing of application caused a difference in dough properties, such as water absorption, mixing time, and tolerance, among the cultivars. Soluble and insoluble polymeric and monomeric protein contents increased with an increase in N amount and timing of application the three Korean wheat cultivars. The effects of N amount and application timing on bread loaf volume (BLV) varied among the cultivars. The BLV of Saekeumkang increased regardless of the N amount and timing of application, but that of Baekkang and Joongmo2008 cultivars was reduced. However, there was a positive correlation between protein content with the addition of N fertilization and BLV. In addition, SDSS, mixing time, and protein fractions were positively correlated with BLV. Since the response of fertilizer conditions was different for each wheat cultivar, it is necessary to build a suitable fertilizing system for each of them. Additionally, since the environment is changing, such as abnormal climate during the maturing period, research is needed to establish appropriate fertilizer conditions for varieties of bread wheat.

• Microbiology and Biotechnology Letters
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• v.26 no.6
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• pp.537-544
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• 1998

The cultivation conditions of transformant Alcaligenes eutrophus AER5 harboring cloned phbC gene for mass production of poly (3-hydroxybutyrate-3-hydroxyvalerate)[P(3HB-3HV)] containing high molar fraction of 3-hydroxyvalerate (3-HV) were investigated. In two-stage batch cultivation, transformant accumulated P(3HB-3HV) containing 52.2 mol% of 3HV compared to 30 mol% of parent strain A. eutrophus H16. The increased 3-HV molar fraction was due to the amplified activity of PHB synthase participating in condensation of 3-HB and 3-HV. To increase efficiency of P(3HB-3HV) accumulation, fructose was added along with precursor compound valerate, and total cell mass and P(3HB-3HV) concentrations remarkably increased, but not 3-HV molar fraction. The effect of magnesium ion showed that P(3HB-3HV) concentration and 3-HV molar fraction were significantly increased upto 6.1 g/L and 71.3 mol% at 0.01 g/L of MgSO$_4$, respectively. The efficiency of several pH adjuster, NaOH, NaOH and (NH$_4$)$_2$SO$_4$, and NH$_4$OH, on total cell mass, p(3HB-3HV) concentration, and 3-HV molar fraction was also compared. To overcome the disadvantage of two-stage cultivation, one-stage intermittent fed-batch cultivation was attempted, such a way 10.0 g/L of fructose was supplied for cell growth at initial 36 hr and then 10.0 g/L of valerate and 5.0 g/L of fructose were applied to induce the accumulation of P(3HB-3HV), consequently, 10.4 g/L of P(3HB-3HV) with 38 mol% of 3-HV fraction could be obtained after 72 hr. These results can be used for elucidating cultivation strategy for mass production of P(3HB-3HV) containing high 3-HV molar fraction using transformant A. eutrophus AER5 harboring cloned phbC gene.

• Microbiology and Biotechnology Letters
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• v.26 no.4
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• pp.323-330
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• 1998

Cyclodextrin glucanotransferase (CGTase) was purified from the culture broth of the Bacillus firmus var. alkalophilus, using ultrafiltration, starch adsorption/desorption, ion-exchange chromatography on DEAE-cellulose and gel filtration on Sephacryl HR-100. The molecular weight of the purified enzyme was determined as 77,000 by SDS-PAGE. The optimum pH and temperature for the CD synthesis were 6.0 and 5$0^{\circ}C$, respectively. The activity of this enzyme was stably kept at the range of pH 6.0～9.5 and up to 5$0^{\circ}C$. However, in the presence of $Ca^{2+}$, the optimum temperature for CD synthesis was shifted 55~6$0^{\circ}C$ and this enzyme was stable up to 6$0^{\circ}C$ because of the stabilizing effect of $Ca^{2+}$. The purified CGTase produced CDs with high conversion yields of 45~51% from sweet potato starch, com starch and amylopectin as substrate, especially, and the product ratio of $\beta$-CD to ${\gamma}$-CD was obtained at range of from 5.8:1 to 8.4:1 according to the kind of substrate. The purified enzyme produced mainly $\beta$-CD without accumulation of $\alpha$-CD during enzyme reaction using various starches as the substrate, indicating that the purified enzyme is the typical $\beta$-CGTase. The purified CGTase produced 25 g/l of CDs from 5.0% (w/v) liquefied com starch and the conversion yield of CDs was 50%, and the content of $\beta$-CD was 84% of total CDs after 8 hours under the optimum reaction condition.ion.

• Journal of Life Science
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• v.27 no.2
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• pp.202-210
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• 2017

As an effort to find a potential biopreservative, we isolated bacterial strains producing bacteriocin from fermented foods. A strain was finally selected and characteristics of the bacteriocin were investigated. The selected strain was identified as Bacillus subtilis E9-1 based on the 16S rRNA gene analysis. The culture supernatant of B. subtilis E9-1 showed antimicrobial activity against Gram-positive bacteria. Subtilisin A, ${\alpha}$-chymotrypsin, trypsin and proteinase K inactivated the antimicrobial activity, which means its proteinaceous nature, a bacteriocin. The bacteriocin activity was fully retained at the pH range from 2.0 to 8.0 and stable at up to $100^{\circ}C$ for 60 min. Solvents such as ethanol, isopropanol and methanol had no effect on the antimicrobial activity at the concentration of 100% but acetone and acetonitrile reduced the activity at up to 100% concentration. Cell growth of four indicator strains was dramatically decreased in dose-dependent manner. Listeria monocytogenes was the most sensitive, but Enterococcus faecium was the most resistant. Bacillus cereus and Staphylococcus aureus showed the medium sensitivity. The bacteriocin showed its antimicrobial activity against B. cereus and L. monocytogenes via bactericidal action. The number of viable cells of L. monocytogenes started to reduce after addition of bacteriocin to the minced beef. The bacteriocin was purified through acetone concentration, gel filtration chromatography and RP-HPLC. The whole purification step led to a 6.82 fold increase in the specific activity and 6% yield of bacteriocin activity. The molecular weight of the purified bacteriocin was determined to be 3.3 kDa by MALDI-TOF/TOF mass spectrometry.

• Journal of the Korean Society of Food Science and Nutrition
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• v.30 no.5
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• pp.894-899
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• 2001

Intakes and excretions of zinc and copper were determined for 8 female adolescents (aged 16.4$\pm$0.5 y; body mass index 20.4$\pm$1.3kg/$m^2$; body fat 33.3$\pm$2.5%; bone mineral density of lumbar spine in L2-L4; 0.96$\pm$0.08g/$\textrm{cm}^2$) when they consumed diets basal and high in calcium for 6 days each. All subjects consumed a basal Ca diet containing 800 mg, Korean RDA level of the subjects, and a high Ca diet containing 1200mg, RDA plus 2 SDs of calcium intake. The diets provided 58% of energy intake as carbohydrate, 25% as fat, and 17% as protein. Food, urine and fecal samples were collected during the last 3 days of each feeding period and were assayed. Mean daily intakes on the basal and high calcium diets, respectively, were 6.57 and 6.37 mg for zinc and 910 and 812 $\mu\textrm{g}$for copper. Fecal excretion of copper and zinc in relation to intake was significantly greater on the high calcium than on the basal calcium diet. Hence, apparent absorption rate was significantly lowered from 98.7% on the basal calcium diet to 97.9% on the high calcium diet for zinc from 66.3% to 56.4% for copper, respectively. Urinary loss of copper was not detectable but that of zinc was 0.38mg on the basal diet and 0.47mg on the high calcium diet. Copper retention was 899$\pm$105$\mu\textrm{g}$/day on the basal calcium diet and 792$\pm$20.8$\mu\textrm{g}$/day on the high calcium diet, and zinc retention was 3.95$\pm$0.91mg/day and 3.11$\pm$0.89mg/day. Thus, copper and zinc retention was significantly decreased on the high calcium diet (p<0.05). Summarizing the results, apparent absorption and retention of zinc and copper were significantly decreased by calcium supplementation. Therefore, it is suggested that interactions among minerals should be considered in determining RDA.