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Dietary Effect of Silk Protein on Ceramide Synthesis and the Expression of Ceramide Metabolic Enzymes in the Epidermis of NC/Nga Mice (실크단백질의 식이 공급이 아토피 피부염 동물 모델 NC/Nga Mice 피부의 세라마이드 함량 및 관련인자 발현에 미치는 영향)

  • Park, Kyung-Ho;Choi, Young-Sim;Kim, Hyun-Ae;Lee, Kwang-Gill;Yeo, Joo-Hong;Jung, Do-Hyun;Kim, Sung-Han;Cho, Yun-Hi
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.5
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    • pp.554-562
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    • 2007
  • Ceramide rich intercellular lipid lamellae are thought to be particularly important in maintaining the structural integrity of epidermal barrier. Ceramide is synthesized de novo by serine palmitoyltransferase (SPT) phospholipid intermediates, serine and palmitic acid persist within the stratum corneum. The ceramide which is synthesized is degraded with fatty acid and sphingosine by degradative enzyme ceramidase. The depletion of ceramide in stratum corneum was reported in the atopic dermatitis. As an effort to search for the dietary source for improving the level of ceramide in epidermis, the dietary effects of various-typed silk protein were compared. Seventy male NC/Nga mice, an animal model of atopic dermatitis, were divided into seven groups: group CA as an atopic control with control diet, group S: 1% crude sericin diet, group F: 1% crude fibroin diet, group PS : peptide pattern of sericin(Mw 5000), group PF: peptide pattern of fibroin (Mw 1500), group AS: manufactured the same as amino acid profile of sericin and group AF: manufactured the same as amino acid profile of fibroin. Ten male BALB/c mice were served as group C (control group) control diet. All mice were fed on diet and water ad libitum for 10 weeks. Dry skin condition was established in group CA as ceramide content was decreased. Despite a marked decrease of mRNA and prorein expression of SPT, enzyme do novo synthesis, ceramide content of group S was dramatically increased by inhibiting the mRNA and protein expression of degradative enzyme ceramidase. However, dietary supplementation of crude silk fibroin protein (group F) and in other groups that were supplemented with either amino acid or peptide type of sericin or fibroin did not increase the level of ceramide. Together, our data demonstrate that dietary supplementation of crude sericin is more effective at improving ceramide level in epidemis of NC/Nga mice.

Antimutagenic and Cytotoxic Effects of Ethanol Extracts from Five Kinds of Seaweeds (다섯 가지 해조류 에탄올 추출물의 항돌연변이 활성 및 암세포 성장억제 효과)

  • Kim, Sung-Ae;Kim, Jin;Woo, Mee-Kyung;Kwak, Chung-Shil;Lee, Mee-Sook
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.4
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    • pp.451-459
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    • 2005
  • The protective effects of ethanol extracts from 5 seaweeds on the mutagenic and cytotoxic damage were evaluated. They were separately extracted using ethanol from dried samples at room temperature, and freeze-dried. The inhibition effects on the mutagenicity in Salmonella assay by Ames test and cancer cell inhibitory effect in HeLa cell, MCF-7 cell and SNU -638 cell by MTT assay were assayed. Seaweed fusiforme, sea tangle and green laver showed strong inhibitory effect against 2-nitrofluorene, sodium azide- or 2-anthramine-induced mutagenicities in Salmonella Typhimurium TA 98 and TA 100 at the level of 2.5 mg ethanol extract per plate. Cancer cell inhibitory effect was shown with all of the seaweed extracts. Green laver, sea mustard, sea tangle and seaweed fusiforme showed strong cytotoxicity against HeLa and MCF-7 cells, with inhibiting by $92\~93\%$ and $89\~92\%$, respectively. These data show that 5 seaweeds tested in this study might be potent functional foods for cancer prevention, and consumption of these seaweeds in adequate amount is recommended.

Effect of Foliar Sprays of CaCl2 for Improving Fruit Quality of (복숭아 )

  • Kim, Ik-Youl;Kim, Mi-Young;Ru, Jong-Ho;Kim, Min;Lee, Yong-Se;Chang, Tae-Hyun
    • Korean Journal of Environmental Agriculture
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    • v.25 no.3
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    • pp.276-283
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    • 2006
  • To evaluate the effectiveness of the foliar spray of calcium on "Baekdo" peach fruit, we carried out experiments in the orchards. The sprays were applied with $CaCl_2\;(Ca:\;400mg.kg^{-1})\;and\;CaCl_2$ with adjuvants (amino acid, $2g.kg^{-1}$; phytic acid, $2ml.kg^{-1}$ and wood vinegar, $2ml.kg^{-1}$) for four times from June 12 through July 4 at weekly intervals. The fruits and leaves were evaluated for Ca content, firmness and incidence of Brown rot caused by Monilinia fructicola at harvest To evaluate fruit quality included Ca content, firmness natural decay during the storage, the fruits were stored at room temperature for 14 days. The Ca content in leaf and fruit flesh at harvest was significantly increased (P=0.05) in $CaCl_2$ + amino acid treatment among $CaCl_2$ treatments. However, there was not significant Ca content in fruit peel. The firmness of flesh increased significantly (P=0.05) in $CaCl_2$ + amino acid treatment. The natural decay (Rhizopus stolonifers) during storage at the room temperature for 14 days, the fruit treated with $CaCl_2$ + wood vinegar exhibited lowest (P=0.05) incidence. Also, the firmness of the fruit during storage was firmer with treated $CaCl_2$ than untreated fruit. In the treatments of $CaCl_2$ + phytic acid and $CaCl_2$ + amino acid, it was possible to reduce incidence of Brown rot caused by M. fructicola most effectively in the field. In addition, inoculation with M. fructicola in fruits was also the most effective treatment for inhibiting disease development in vitro. These results suggested that the foliar spray of $CaCl_2$ with adjuvants increased the content of Ca and firmness of the fruits positively. It also inhibited the natural decay and the Brown rot effectively.

Effect of Inoculum Concentration of Rhizoctonia solani and Pythium ultimum Causing Damping-off of Pepper and Cucumber on the Efficacy of the Mixture of Etridiazole and Thiophanate-methyl (Rhizoctonia solani와 Pythium ultimum의 접종 농도가 etridiazole과 thiophanate-methyl 합제의 모잘록병 방제 효과에 미치는 영향)

  • Kim, Hyung-Jo;Jang, Ho-Sun;Lee, Soo-Min;Kim, Joo-Hyung;Shin, Jin-Ho;Kim, Heung-Tae
    • The Korean Journal of Pesticide Science
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    • v.14 no.1
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    • pp.65-71
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    • 2010
  • This study was performed to investigate the inhibiting activity of etridiazole and thiophanate-mthyl on mycelial growth of Rhizoctonia solani and Pythium ultimum and the effect of inoculum density of each pathogen on the control efficacy of the mixture of etridiazole and thiophanate-methyl in a seedling assay test. In mycelial growth inhibition test, $EC_{50}$ values of etridiazole and thiophanate-methyl against R. solani were 15.87 and 9.34 ${\mu}g\;mL^{-1}$, while those were 0.2 and more than $500\;{\mu}g\;mL^{-1}$ against P. ultimum, respectively. Controlling activity of the mixture of etridiazole and thiophanate-methyl against damping-off of pepper and cucumber, caused by R. solani and P. ultimum was tested in a greenhouse. With the inoculation of R. solani, disease incidences of seedling in pepper were 73% and 95% at 0.5% and 1.0% of inoculum concentration, while in cucumber those were 55% and 62% at 1.0% and 2.0%. When P. ultimum was inoculated into soil by 2.0% of inoculum concentration, those in pepper and cucumber were 66.7% and 96.8%, respectively. The efficacy of the mixture was somehow affected by the concentration of R. solani. While each control value of the mixture was 94.4% and 90.7% in pepper and cucumber at low inoculum concentration (0.05%), the efficacy of the fungicide decreased in pepper and cucumber by 70.7% and 72.9% at high concentration of R. solani (0.1% in pepper and 0.5% in cucumber). However, the control value of the mixture was 100% in pepper and cucumber, irrespective of the inoculum concentration of P. ultimum, however, the increase of inoculum concentration in soil did not result in the decrease of the fungicide efficacy.

Use of Antimicrobial Agents for the Treatment of Inpatients in Chonbuk National University Hospital (전북대학교병원 입원환자에서의 항생제 사용 실태)

  • Song, Jae Ho;Kim, Jung Soo
    • Pediatric Infection and Vaccine
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    • v.7 no.2
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    • pp.225-232
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    • 2000
  • Purpose : The use antimicrobial agents is one of the important strategies for the treatment and prophylaxis of microbial infections. But injudicious abuse and misuse of antimicrobial agents is problem to add an extra weight on medical fee, increase of resistant bacteria and side effects according to the antibiotic use. This study was performed to establish the pertinent use of antimicrobial agent in Chonbuk National University Hospital(CNUH). Characteristics of antibiotics use was analysis by reviewing the medical records of patients admitted to CNUH during the period of May 1998. Methods : One thousand eight hundred and thirty three patients were enrolled in this study(medical division 1,014 cases, surgical division 819 cases). Medical records were retrospectively reviewed to classify the rate of antibiotics use, name of antibiotics used, appropriateness of antibiotics use. Results : The overall rate of antibiotic usage in CNUH was 67.2%(1,231/1,833), showing higher rate in surgical division(89.6%) compare to that of medical division(49.0%). Among 1,231 patients to whom antimicrobial agents were given, only 125(10.2%) were treated with single antimicrobial agents. 311(25.3%) were treated with two antimicrobial agents, and 795(64.5%) patients received 3 or more antibiotics. ${\beta}$-lactams(56.4%) were most frequently used followed by aminoglycosides(35.3%), the others(4.9%) and quinolons(3.4%). Amoxicillin-clavulanate was the mostly commonly used antibiotics followed by amoxicillin and unasyn. Prophylactic use of antibiotics was carried in seven hundred six patients(57.4%), mostly in surgical division, which can be considered somewhat inappropriate in the initiation time and duration of antibiotic use. Conclusion : Importance of monotherapy and appropriate prophylactic antibiotic use should be emphasized. Strategies of antibiotics use, such as restriction of drug use, continuous monitoring system, flow sheet system should be considered to reduce antibiotics use and establish the appropriate use of antibiotics as well as inhibiting the occurrence of resistant strains.

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Comparison of Antioxidant, Cytotoxicity and Flavonoid Content of Stachys sieboldii Miq. vs. Lycopus lucidus Turcz. Leaf Extracts (초석잠 및 택란 잎 추출물의 항산화 및 세포독성 활성과 총 플라보노이드 함량 비교)

  • Na, Eun;Lee, Jung Woo;Lim, Sun Young
    • Journal of Life Science
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    • v.30 no.2
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    • pp.147-155
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    • 2020
  • In this study, the antioxidant and cytotoxic effects and the flavonoid contents of leaf extracts from Stachys sieboldii Miq. and Lycopus lucidus Turcz. were compared. The flavonoid contents of the acetone + methylene chloride (A+M) and methanol (MeOH) extracts of L. lucidus Turcz. leaves were 55.7 and 233.2 mg/g, respectively. In a DPPH assay, A+M and MeOH extracts from L. lucidus Turcz leaves had a greater scavenging effect than those of S. sieboldii Miq. leaves (p<0.05). In an ABTS assay, MeOH extracts from S. sieboldii Miq. and L. lucidus Turcz (0.5 mg/ml concentration) leaves had scavenging effects of 85% and 91%, respectively (p<0.05), suggesting that both of the MeOH extracts had greater scavenging effects than both A+M extracts. In a 120 min ROS production assay, all tested extracts decreased the cellular ROS production induced by H2O2 compared to that produced by exposure to the extract-free control. The MeOH extract from L. lucidus Turcz leaves had a greater inhibitory effect on cellular ROS production (p<0.05). Treatment with A+M and MeOH extracts from both S. sieboldii Miq. and L. lucidus Turcz. leaves showed a dose-dependent increased cytotoxicity against the growth of AGS, HT-29 cancer cells, and HT-1080 (p<0.05). Both A+M extracts had a greater inhibitory effect on the growth of all cancer cells than both MeOH extracts. These results suggest that the MeOH extract of L. lucidus Turcz. leaves is effective in scavenging free radicals and inhibiting cellular oxidation, while the A+M extract inhibits proliferation of three types of cancer cell.

The Cross-talk Mechanisms of Constitutive Androstane Receptor (CAR) in the Regulation of its Activity, Energy Metabolism, Cellular Proliferation and Apoptosis (Constitutive Androstane Receptor (CAR)의 활성, 에너지 대사 및 세포의 증식과 사멸의 조절에 대한 CAR의 cross-talk 기전)

  • Min, Gyesik
    • Journal of Life Science
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    • v.30 no.2
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    • pp.211-220
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    • 2020
  • The activity of CAR can be regulated not only by ligand binding but also by phosphorylation of regulatory factors involved in extracellular signaling pathways, cross-talk interactions with transcription factors, and the recruitment, degradation, and expression of coactivators and corepressors. This regulation of CAR activity can in turn have effects on the control of diverse physiological homeostasis, including xenobiotic and energy metabolism, cellular proliferation, and apoptosis. CAR is phosphorylated by the ERK1/2 signaling pathway, which causes formation of a complex with Hsp-90 and CCRP, leading to its cytoplasmic retention, whereas phenobarbital inhibits ERK1/2, which causes dephosphorylation of the downstream signaling molecules, leading to the recruitment to CAR of the activated RACK-1/PP2A components for the dephosphorylation, nuclear translocation, and the transcriptional activation of CAR. Activated CAR cross-talks with FoxO1 to induce inhibition of its transcriptional activity and with PGC-1α to induce protein degradation by ubiquitination, resulting in the transcriptional suppression of PEPCK and G6Pase involved in gluconeogenesis. Regulation by CAR of lipid synthesis and oxidation is achieved by its functional cross-talks, respectively, with PPARγ through the degradation of PGC-1α to inhibit expression of the lipogenic genes and with PPARα through either the suppression of CPT-1 expression or the interaction with PGC-1α each to induce tissue-specific inhibition or stimulation of β-oxidation. Whereas CAR stimulates cellular proliferation by suppressing p21 expression through the inhibition of FoxO1 transcriptional activity and inducing cyclin D1 expression, it suppresses apoptosis by inhibiting the activities of MKK7 and JNK-1 through the expression of GADD45B. In conclusion, CAR is involved in the maintenance of homeostasis by regulating not only xenobiotic metabolism but also energy metabolism, cellular proliferation, and apoptosis through diverse cross-talk interactions with extracellular signaling pathways and intracellular regulatory factors.

Effect of Dietary Selenium on the Colon Carcinogenesis in Male ICR Mice

  • Cho, Min-Haeng;Kim, Jun-Hyeong;Hue, Jin-Joo;Kang, Bong-Su;Park, Hyun-Ji;Nam, Sang-Yoon;Yun, Young-Won;Kim, Jong-Soo;Jeong, Jae-Hwang;Lee, Beom-Jun
    • Journal of Food Hygiene and Safety
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    • v.25 no.3
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    • pp.269-277
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    • 2010
  • Selenium is an essential micronutrient for normal body function and functions as an essential constituent of selenoproteins. This study was carried out to investigate effect of selenium on the formation of colonic aberrant crypt foci (ACF) and tumor formation in a mouse model. Five-week old ICR mice were acclimated for one week and fed different selenium diet (0.02, 0.1, and 0.5 ppm) for 12 weeks. Animals received three intraperitoneal injections of azoxymethane (10 mg/kg B.W. in saline for 3 weeks), followed by 2% dextran sodium sulfate in the drinking water for a week. There were four experimental groups, including a normal control group and three different selenium levels groups. After sacrifice, the total numbers of aberrant crypt (AC) and ACF were measured in the colonic mucosa after methylene blue staining. The number of tumors was noted for tumor incidence. Liver selenium concentration was measured using ICP-AES method. Gutathione peroxidase (GPx) activity was determined using a GPx assay kit in the liver and colon. TUNEL assay and proliferating cell nuclear antigen (PCNA) staining were performed to examine the cell apoptosis and cell proliferation, respectively. Immunohistochemistry of $\beta$-catenin was also performed on the mucous membrane tissue of colon. The activity of GPx in the liver and colon was decreased in the selenium-deficient diet group while it was increased in the selenium-overloaded diet group. Apoptotic positive cells were increased in the selenium-overloaded diet group but decreased in the selenium-deficient diet group. PCNA staining area was decreased in the selenium-overloaded diet group. In addition, the $\beta$-catenin protein level in the selenium-deficient diet group was increased but decreased in the selenium-overloaded diet group. These results indicate that dietary selenium might exert a modulating effect on colon cancer by inhibiting the development of ACF and colon tumor formation in this mouse model.

Characterization of Microsomal $Ca^{2+}$ Uptake in Tomato Root Tissues (토마토 뿌리조직에서 분리한 마이크로솜의 $Ca^{2+}$ 흡수 특성)

  • Cho, Kwang-Hyun;Kim, Young-Kee
    • Applied Biological Chemistry
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    • v.42 no.2
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    • pp.116-122
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    • 1999
  • In order to characterize the property of $Ca^{2+}$ transport in plant cell, microsomes were prepared from the roots of tomato and microsomal $^{45}Ca^{2+}$ uptake was measured. When 1 mM vanadate, a selective inhibitor of P-type ATPases, 50 mM $NO_3^-$, a specific inhibitor of vacuolar $H^{+}-ATPase$, and both of these inhibitors were treated, the microsomal $^{45}Ca^{2+}$ uptakes were inhibited by 20, 33 and 47%, respectively. The inhibitory effects of these two inhibitors were investigated by using a protonophore, gramicidin. When the chemical gradient of $H^{+}$ was relieved by gramicidin, the uptake was decreased by 30%, implying the presence of $Ca^{2+}/H^+$ antiporter in the microsomal membrane. In the $^{45}Ca^{2+}$ uptake experiment, the effect of gramicidin was independent of vanadate-induced inhibition. However, when the activity of vacuolar $H^{+}-ATPase$ was inhibited by $NO_3^-$, the effect of gramicidin was severely decreased. Meanwhile, thapsigargin, a specific antagonist of ER/SR-type $Ca^{2+}-ATPase$, inhibited the microsomal $^{45}Ca^{2+}$ uptake and the maximum inhibitory effect was obtained at $10\;{\mu}M$. The effect of thapsigargin was blocked by $NO_3^-$ and gramicidin, but not by vanadate. These results imply that vanadate directly inhibits the activity of $Ca^{2+}-ATPase$; however, $NO_3^-$ and thapsigargin block the activity of $Ca^{2+}/H^+$ antiporter by inhibiting the vacuolar $H^{+}-ATPase$. In conclusion, the microsomal $^{45}Ca^{2+}$ uptakes are mediated by two major enzymes, $Ca^{2+}-ATPase$ and $Ca^{2+}/H^+$ antiporter in tomato root tissue.

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Oocyte-sperm Binding Assay (OSBA) Technique for Rapid Q/C of IVF Culture Condition (체외수정용 배양조건의 신속한 Q/C를 위한 정자-난자 결합분석법(OSBA) 개발)

  • 정구민;신영수
    • Korean Journal of Animal Reproduction
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    • v.25 no.2
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    • pp.163-169
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    • 2001
  • OSBA(oocytes-sperm binding assay) is a tool developed for rapid test of optimal condition of IVF medium and protein source by binding ability of mouse sperm and egg. Mouse oocyte-cumulus complexes were prepared by removing of the cumulus cells with 0.1% hyaluronidase. 10$\pm$2 oocytes per 30 ${mu}ell$ medium drop were inseminated with 3 ${mu}ell$ sperm suspension and were cultured f3r 3 hours and 24 hours, respectively. And the oocytes were recovered gently and the No. of sperm bound on oocytes were counted. In the Exp. 1, the ratio of oocytes bound with one sperm at least were 60.2%(50/83), 2%(2/77) and 100%(79/79) in the medium with no protein, FBS(15%, v/v) and BSA(0.4%. w/v), respectively, Fetal bovine serum(FBS) seriously inhibited sperm binding on oocyte, although bovine serum albumin(BSA) promoted the binding ability. The inhibiting effect of FBS was dependent on the concentration of FBS. The sperm binding ability according to oocyte maturity was tested in the Exp. 2. There was no significant difference between Met. II (mature) and Met. I (intermediate mature) oocytes in the number of oocytes bound with sperm and the number of sperm bound on oocytes. Finally, in Exp. 3, two batches of Ham's F10 medium with good and poor quality by OSBA were tested (The ratios of embryos developed from PN 1-cell stage to hatched blastocyst; 25% vs. 70%). In the medium with good quality, sperm binding ability was significantly increased (P < 0.05). The ratio of oocytes bound with one sperm at least was 66% and 90% in the medium with poor and good quality, respectively. Conclusively, It was possible to test IVF medium condition rapidly and easily by OSBA.

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