• Title/Summary/Keyword: stress protein

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A MOLECULAR BIOLOGIC STUDY ON BIOCOMPATIBILITY OF METALLIC DENTAL MATERIALS USED FOR CHILDREN WITH CULTURED HUMAN GINGIVAL FIBROBLASTS (인체 섬유모세포(HGF-1) 배양에서 소아용 치과금속재의 세포친화성에 대한 분자생물학적 연구)

  • Kim, Ju-Mi;Jeong, Tae-Sung;Kim, Shin
    • Journal of the korean academy of Pediatric Dentistry
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    • v.29 no.2
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    • pp.243-254
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    • 2002
  • For the purpose of evaluating the biocompatability of 3 kinds of metallic materials frequently used in pediatric dentistry (stainless steel crown, orthodontic band, orthodontic wire), cellular and molecular studies, including cell growth and proliferation, screening of cell death with determination of types whether necrosis or apoptosis and changes in expressions of related signaling molecules were examined, using cultured human gingival fibroblasts (HGF-1), HGF-1 was cultured in Dulbecco's modified Eagle's medium. among which the 3rd to 6th generations of HGF-1 were used. The specimen were divided into stainless steel crown (R), band (B) and wire (W). The immunocytochemical study was done for the detection of anti-PCNA (proliferating cell nuclear antigen) labeling. With extracted protein, western blot was done for the detection of ERK1/2, JNK, and p38, using individual antibodies. Cultured cells proliferated, remarkably till 7 day and slightly at 11 day. There was no statistical significance in the counts of proliferating HGF-1 between control and experimental groups (p>0.05). Relative growth rates were no statistically significant difference between control and experimental groups (p>0.05). PCNA labeling indexes showing similar patterns in control and experimental groups. The expressions of ERK1 and ERK2, p38 were similar in control and experimental groups. The expression of JNK increased at 1st day, slightly decreased at 4th day and markedly increased at 7th and 11 day. Although the patterns of control and experimental groups were similar, the increased expressions of JNK at late period suggest a possible stress due to inhibited cell growth and proliferation, and worse culture condition. Conclusively, the 3 kinds of metal specimens used in this study did not induce cellular and molecular hazards during short term culture of HGF-1. But, for the better clinical stability, the establishment of long period culture and animal experiment was thought necessary.

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Antioxidant activity and physicochemical composition of fermented Vigna angularis using Bacillus subtilis KCCM 11965P (Bacillus subtilis KCCM 11965P를 이용한 붉은팥 발효물의 항산화 활성 및 이화학성분 분석)

  • Jeong, Kyung Ok;Oh, Keun Su;Moon, Kwang Hyun;Kim, Dae Geun;Im, So Yeon;Lee, Eun Ji;Kim, Na Ri;Kim, Wook;Kim, Hae Jin;Lee, Jeong Ho
    • Food Science and Preservation
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    • v.24 no.7
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    • pp.975-982
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    • 2017
  • Health of human intestine has deteriorated due to excessive stress and western diet. In this study, Vigna angularis was fermented by Bacillus subtilis KCCM11965P in order to develop prebiotic resource for improving bowel movement. The contents of ash and crude protein were $3.35{\pm}0.04%$ and $21.1{\pm}0.19%$ respectively. Vigna angularis extract (1, 3, 5%) were incubated with 3% (v/v) Bacillus subtilis KCCM 11965P for 0, 24, 48, and 72 h. Total bacterial numbers showed that the combination of 3% powdered seeds and 72 h incubation time was optimum condition for this experiment. Total polyphenol content increased from $0.18{\pm}0.010mg/mL$ in pre-incubation to $0.23{\pm}0.007mg/mL$ in post-incubation with the condition mentioned above. DPPH radical scavenging activity also increased from $36.1{\pm}6.0%$ to $63.6{\pm}5.2%$. Analysis of protease activity showed $2.69{\pm}0.003unit/mL$ in combination of 5% powdered seeds and 72 h incubation time. Amylase activity increased from $1.0{\pm}0.1unit/mL$ in pre-incubation to $26.0{\pm}0.2unit/mL$ in post-incubation. The analysis of free amino acids after incubation with Bacillus subtilis KCCM 11965P showed that leucine increased from 5.22 mg/L to 67.59 mg/L and tyrosine, one of non-essential amino acid also increased 10.08 mg/L to 259.35 mg/L by incubation with 5% powdered seeds. Most of organic acid were reduced by incubation for 72 h. These results suggest that Vigna angularis could be utilized most as a prebiotic resources.

Anti-oxidative and Neuroprotective Activities of Pig Skin Gelatin Hydrolysates (돈피젤라틴 효소분해물의 항산화 활성 및 신경세포보호효과)

  • Kim, Dong Wook;Park, Kimoon;Ha, Goeun;Jung, Ju Ri;Chang, Ounki;Ham, Jun-Sang;Jeong, Seok-Geun;Park, Beom-Young;Song, Jin;Jang, Aera
    • Food Science of Animal Resources
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    • v.33 no.2
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    • pp.258-267
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    • 2013
  • This study was conducted to determine the antioxidative and neuroprotective effect of pig skin extracts (PS) and pig skin gelatin hydrolysates (LPS) using a human neuroblastoma cell line (SH-SY5Y). The extraction yield of PS was 3 fold higher than that of LPS. The protein content of PS was about 10 fold higher than that of LPS (p<0.05). Also LPS increased antioxidative activity dose dependently, and the activity was significantly higher than PS at all concentration (p<0.05). DPPH radical scavenging activity of LPS at 50 mg/mL was 92.97%, which was similar to $1{\mu}M$ vitamin C as a positive control. ABTS radical scavenging activity of LPS (20 mg/mL) was 89.83% and oxygen radical absorbance capacity of LPS at 1 mg/mL was $141.39{\mu}M$ Trolox Equvalent/g. No significant change of human neuroblastoma cells was determined by MTT test. Cell death by oxidative stress induced by $H_2O_2$ and amyloid beta 1-42 ($A{\beta}_{1-42}$) was protected by LPS rather than PS. Acetylcholine esterase was significantly inhibited, by up to 33.62% by LPS at 10 mg/mL. Therefore, these results suggest that pig skin gelatin hydrolysates below 3 kDa have potential to be used as anti-oxidative and neuroprotective functional additives in the food industry, while further animal test should be determined in the future.

Effects of Dietary Supplementation with Allium hookeri Root on Hepatic Enzyme Contents in Streptozotocin-induced Diabetic Rats (삼채 보충식이가 당뇨흰쥐 간의 항산화효소 함량에 미치는 영향)

  • Kim, Myung-Wha
    • Journal of the East Asian Society of Dietary Life
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    • v.27 no.4
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    • pp.399-407
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    • 2017
  • The purpose of this study was to examine the effect of Allium hookeri (AH) root on hepatic antioxidative enzyme contents in streptozotocin (STZ)-induced rats. Diabetes mellitus was induced in male Sprague-Dawley rats through injection of STZ dissolved in citrate buffer into tail veins at a dose of 45 mg/kg body weight. Sprague-Dawley rats were fed an AIN-93 recommended diet, and the experimental groups were fed a modified diet containing 5% and 10% of AH root powder for 4 weeks. The experimental groups were divided into four groups: a normal control (N-control), STZ-control, STZ-AH 5%, and STZ-AH 10% supplemented groups. The STZ-AH 5% group showed a significant increase in liver glycogen compared to the STZ-control group. Muscle glycogen and liver protein contents significantly increased in the AH-supplemented groups compared to the STZ-control group. The liver malondialdehyde content of the AH-supplemented group was significantly lower than that of the STZ-control group. Xanthine oxidase content was significantly reduced in all experimental groups. Glutathione-S-transferase content was significantly elevated in the AH-treated groups compared to the STZ-control group. Superoxide dismutase content was not significantly different among the experimental groups. Catalase content was significantly higher in the STZ-AH 10% group compared to the STZ-control group. These results show that supplementation with AH root may be useful for diabetic therapy and damage from oxidative stress.

Antioxidant and Anti-inflammatory Activity of Ethanol Extract of Malus micromalus Makino in Jeju Island (제주도 자생 제주아그배 (Malus micromalus Makino) 추출물의 항산화와 항염증 활성)

  • Lee, Ju-Yeop;Kang, Min-Chul;Lee, Jung-A;Ko, Kwang-Hyo;Kim, Bong-Seok;Han, Jong-Heon;Kim, Se-Jae;Kim, Gi-Ok
    • KSBB Journal
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    • v.24 no.4
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    • pp.327-333
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    • 2009
  • The antioxidant and anti-inflammatory activities of ethanol extract of Malus micromalus were studied in vitro. Ethanol extract of M. micromalus showed scavenging effects on 1,1-diphenyl-2-picrylhydrazyl (DPPH) and nitric oxide (NO) radicals. In addition, ethanol extract of M. micromalus inhibited the generation of superoxide anion ($O_2^-$) radical and uric acid by xanthine oxidase. We also investigated the effect of ethanol extract of M. micromalus on NO production in a lipopolysaccharide (LPS)-stimulated murine macrophage RAW 264.7 cells. Ethanol extract of M. micromalus significantly inhibited NO production and this inhibitory action was not due to the cytotoxicity. The expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) was markedly down-regulated by ethanol extract of M. micromalus. These results indicate that the inhibitory action of ethanol extract of M. micromalus on NO production in LPS-stimulated macropages might be due in part to abrogation of iNOS and COX-2 protein induction. Taken together, this study suggests that ethanol extract of M. micromalus could contribute to the chemoprevention and therapy of oxidative stress and inflammation.

Effect of Black Garlic and Herb Formulas on Lipid Profiles and Antioxidant Status in Rats by Interval Running Training (간헐적 운동부하에서 흑마늘과 생약재의 복합처방이 흰쥐의 체내 지질 성분 및 항산화에 미치는 영향)

  • Lee, Soo Jung;Kim, In Sung;Lee, Hye Jin;Oh, Soo Jeong;Shin, Jung Hye;Kim, Jeong Gyun;Sung, Nak Ju
    • Journal of Life Science
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    • v.23 no.12
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    • pp.1436-1444
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    • 2013
  • To improve the functionality of black garlic drinks, black garlic extract (5%) and five herb extracts (1%) were mixed in 70:30 (v/v) ratios as BHF1, and BHF2 was prepared using a 3X concentration of BHF1. After the black garlic and herb formulas (BHFs) were administered over the course of five weeks in rats by interval running training, the lipid profiles and the antioxidant enzyme activities were tested. The total phenolic content of the BHFs were significantly higher in BHF2 than they were in BHF1, and their antioxidant activities were dependent upon the total phenolic content. No significant difference was found in the total serum protein levels among the rats in the Ex-con group by interval running training and the rats in the BHFs-fed groups. However, the albumin level was significantly higher in the Ex-BHF2 to Ex-con group. AST and ALT activities significantly decreased in the BHFs-fed groups compared to the Ex-con group. In terms of changes in the serum lipid profiles, no significant difference was found between the specimens that underwent interval running training and those that did not undergo interval running training. Triglyceride levels, total cholesterol, LDL-C, and HTR levels in the serum were significantly decreased in the Ex-BHF2 to Ex-con group. No significant difference was found in the total lipid levels in the livers of the BHFs-fed groups and the Ex-con group. The triglyceride levels and total cholesterol levels in the Ex-BHF2 group were significantly lower compared to another group. Hepatic catalase activity was significantly increased in the Ex-BHF2 group, but SOD and GSH-px activities were significantly increased as the concentration of the BHF. The antioxidant enzyme activities by supplementation of BHFs increased; thus, three intakes of BHF each day could improve antioxidant status against different types of oxidative stress.

Comparison of Blood Physiology in Juvenile Black Seabream (Acanthopagrus schlegeli) Reared in Converted Freshwater from Seawater and Seawater from Freshwater (해수사육에서 담수사육으로, 담수사육에서 해수사육으로 전환된 감성돔 치어, Acanthopagrus schlegeli의 혈액생리학적 비교)

  • Chang Young Jin;Min Byung Hwa;Chang Hae Jin;Hur Jun Wook
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.35 no.6
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    • pp.595-600
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    • 2002
  • Comparison of blood properties in juvenile black seabream (Acanthopans schlegeli) between transfer from seawater to freshwater (sFW) and transfer from freshwater to seawater (fSW) were investigated for 60 days. Plasma cortisol levels in sFW were significantly increased from 34.2 $\pm$ 28.6 ng/mL at the beginning to 365.6 $\pm$ 136.0 ng/mL at 3 hours, and decreased to the beginning level at 24 hours. However, in fSW, no significant differences in the cortisol levels were recognized throughout experimental period. No significant difference was found in the glucose levels between sFW and fSW. The transfer from seawater to freshwater of juvenile black seabream resulted in reduced $Na^{+}\;and\; Cl^{-}$ concentrations for 24 hours (decreasing from 165.3 $\pm$ 2.5, 146.0$\pm$ 2.2 to 118.3 $\pm$ 12.3, 78.0$\pm$ 7.0 mEq/L, respectively), but these were completely recovered the beginning levels at 30 days. Total protein and AST showed no significant differences between the two rearing conditions, while ALT was markedly elevated at 3 hour in sFW and at 24 hours in fSW. Ht in sFW was increased from 18.5 $\pm$ $0.6\%$ at the beginning to 25.3 $\pm$ $4.0\%$ at 12 hours, and was decreased to the beginning level at 24 hours. Ht, RBC and Hb in fSW were also significantly higher at 12 hours, but recovered to their initial levels at 24 hours. All fish were dead until 50 days in sFW while survival rate in fSW was $85\%$ at the end of experiment.

Physicochemical Properties and Physiological Activities of Rhus verniciflua Stem Bark Cultured with Fomitella fraxinea (장수버섯 균사체가 배양된 옻피의 이화학적 특성 및 생리활성)

  • Choi, Han-Seok;Kim, Bo-Hyun;Yeo, Soo-Hwan;Jeong, Seok-Tae;Choi, Ji-Ho;Park, Hyo-Suk;Kim, Myung-Kon
    • The Korean Journal of Mycology
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    • v.38 no.2
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    • pp.172-178
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    • 2010
  • The contents of proximate composition, free amino acids and phenolic acids in the Fomitella fraxinea cultivated-Rhus verniciflua stem bark(FRVSB), and its adipogenesis effect were investigated. The proximate composition(%) of FRVSB was as follows: moisture(7.64), ash(6.30), crude fat(3.86), crude protein(3.59) and sugar(not detected); while Rhus verniciflua stem bark(RVSB) contained 1.64, 8.09, 7.28, 6.48 and 5.39, respectively. The total free amino acids concentration was 97.41 mg% in FRVSB and 71.91 mg% in RVSB. Phosphoserine(55.06 mg%), ammonia(17.84mg%) and aspartic acid(6.05mg%) were predominant amino acids. The content of total phenolic acids was 422.89 ppm in ethanol extract and 283.86 ppm in water extract, with syringic and gallic acid as the main component. The FRVSB extracts showed a potent free radical scavenging activity for DPPH(2,2-diphenyl-1-picrylhydrazyl hydrate) with $IC_{50}$ of $28.54\;{\mu}g$(EtOH) and $54.70\;{\mu}g$(water), respectively, whereas $IC_{50}$ value of gallic acid was $1.84\;{\mu}g$. The protective effect of both ethanol and water extract the extracts against UV-induced oxidative stress in NIH3T3 was observed. The water extracts of FRVSB may promote adipogenesis in 3T3-L1 cells.

Extract from Eucheuma cottonii Induces Apoptotic Cell Death on Human Osteosarcoma Saos-2 Cells via Caspase Cascade Apoptosis Pathway (Eucheuma cottonii 추출물에 의한 인체 골육종암 Saos-2 세포의 자가사멸 유도)

  • Kang, Chang-Won;Kang, Min-Jae;Kim, Kyong Rok;Kim, Nan-Hee;Seo, Yong Bae;Kang, Keon-Hee;Kim, Sang-Ho;Kim, Gun-Do
    • Journal of Life Science
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    • v.26 no.2
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    • pp.147-154
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    • 2016
  • Osteosarcoma (OS) is the most common and malignant bone tumors. Although many types of resection surgery and experimental agents were developed, median survival and clinical prognosis are poorly investigated. Recently, several researches have reported that Eucheuma cottonii has potent as protective effects of coal dust-induced lung damage via inhibition of malondialdehyde (MDA) and oxidative stress in bronchoalveolar lavage fluids (BALF). However, anti-cancer effects and specific molecular mechanism of extract from Eucheuma cottonii (EE) has not been clearly studied yet. This study evaluated that anti-cancer potential of EE in human osteosarcoma Saos-2 cells. EE indicated cytotoxicity on Saos-2 cells in a dose-dependent manner. Morphological degradation and nucleic condensation were also observed under the EE treatment. However, it did not significantly affect on non-cancerous kidney HEK-293 cells under the same concentration which is shown cytotoxicity on Saos-2 cells. The phosphorylation of Fas-Associated Death Domain (FADD) and expression of cleaved caspase-8, -7 and -3 were upregulated in a dose-dependent manner. In immunofluorescence staining, expression level of Fas and cleaved PARP were upregulated by EE treatment. Furthermore, treatment of EE induces upregulation of sub G1 phase by flow cytometry analysis. The results demonstrated that EE has a therapeutic potential against osteosarcoma via FADD mediated caspase cascade apoptosis signal pathway.

Construction and In vitro Study of a Prx 6/Luc Vector System for Screening Antioxidant Compounds in the Transgenic Mice (항산화반응을 유발하는 물질의 검색에 적용할 수 있는 형질전환 마우스 생산을 위한 새로운 Prx 6/Luc 벡터시스템의 제조 및 폐암세포주에서 반응성 확인)

  • Lee, Young Ju;Nam, So Hee;Kim, Ji Eun;Hwang, In Sik;Lee, Hye Ryun;Choi, Sun Il;Kwak, Moon Hwa;Lee, Jae Ho;Jung, Young Jin;An, Beum Soo;Hwang, Dae Youn
    • Journal of Life Science
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    • v.23 no.2
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    • pp.167-174
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    • 2013
  • Peroxiredoxin 6 (Prx 6) is a member of the thiol-specific antioxidant protein family, which may play a role in protection against oxidative stress and in regulating phospholipid turnover. The aim of this study was to determine whether a human Prx 6/Luc vector was stably expressed and responded to antioxidants in a lung cell line (NCI-H460). To achieve this, the luciferase signal, hPrx 6 mRNA expression, and superoxide dismutase (SOD) activity were measured in transfectants with a hPrx 6/Luc plasmid after treatment with four antioxidant extracts, including Korea white ginseng (KWG), Korea red ginseng (KRG), Liriope platyphylla (LP), and red Liriope platyphylla (RLP). First, the hPrx 6/Luc plasmid was successfully constructed with DNA fragments of human Prx 6 promoter, amplified by PCR using genomic DNA isolated from NCI-H460 cells, and cloned into the pTransLucent reporter vector. The orientation and sequencing of the hPrx 6/Luc plasmid were identified with restriction enzyme and automatic sequencing. A luciferase assay revealed significant enhancement of luciferase activity in the four treatment groups compared with a vehicle-treated group, although the ratio of the increase was different within each group. The KRG- and LP-treated groups showed higher activity than the KWG- and RLP-treated groups. Furthermore, the luciferase activity against RLP occurred roughly in a dose-dependent manner. However, the level of endogenous hPrx 6 mRNA did not change in any group treated with the four extracts. The SOD activity was in agreement with the luciferase activity. Therefore, these results indicate that the hPrx 6/Luc vector system may successfully express and respond to antioxidant compounds in NCI-H460 cells. The data also suggest that the Prx 6/Luc vector system may be effectively applied in screening the response of hPrx 6 to antioxidant compounds in transgenic mice.