• Title/Summary/Keyword: Biomarker protein

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Characterization of Bovine Brucellosis in Korean Native Cattle by Means of Immunohistochemistry and Proteomics (면역조직 화학법 및 단백질체 변화 분석을 통한 한우에서 발생한 브루셀라증의 특성)

  • Jang, Seong-Jun;Do, Sun-Hee;Ki, Mi-Ran;Hong, Il-Hwa;Park, Jin-Kyu;Cho, Yu-Jeong;Park, Sang-Joon;Kim, Tae-Hwan;Kwak, Dong-Mi;Jeong, Kyu-Shik
    • Journal of Life Science
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    • v.20 no.2
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    • pp.153-160
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    • 2010
  • This study was conducted to examine the utilization of immunohistochemistry using the bovine anti-brucella immunoglobulin G (IgG) antibody in the diagnosis of brucellosis and to develop a functional biomarker relation for the progress of the disease. Anti-brucella IgG antibody was purified from the affected bovine serum using an affinity chromatography. We performed our investigation on 17 cases of brucellosis and 19 control cases with negative Rose-Bengal test results. Our purified anti-brucella IgG antibody showed a positive immunoreactivity in cytoplasmic hepatocytes of the centrilobular region, and glomeruli and tubular epithelium of the kidney. The protein pattern of the affected liver versus control was analyzed by two-dimensional electrophoresis, showing a different expression pattern of proteins between the two. Five protein spots were up-regulated and another were five down-regulated in the brucellosis liver. Significant upregulaton of catalase and 3-hydroxyacyl-CoA dehydrogenase might be due to a compensatory reaction in response to the endotoxic shock of brucella. In conclusion, the anti-brucella IgG antibody may be a good tool for discriminative diagnosis of the affected tissues and proteomics data suggest new target proteins underlying a possible pathogenic mechanism of brucellosis.

Stress Evaluation to Heavy Metal Exposure using Molecular Marker in Chironomus riparius (분자지표 유전자 발현을 통한 Chironomus riparius 중금속 노출 스트레스 평가)

  • Kim, Won-Seok;Park, Kiyun;Kwak, Ihn-Sil
    • Korean Journal of Ecology and Environment
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    • v.53 no.2
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    • pp.165-172
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    • 2020
  • Heavy metals are common pollutants in the freshwater environment and have toxicological effect in habitat organisms. The heavy metals highly accumulated in sediment and organism, and observed various physiological responses. In this study, we investigated the molecular response to heavy metal toxicity (Al, Aluminum; Cr, Chromium; Cu, copper; Mn, Manganese; Zn, Zinc) through expression of heat shock protein 40, 70, 90 (HSP40, 70, 90), cytochrome 450 (CYP450), Glutathione S-transferase (GST) and Serine-type endopeptidase (SP). HSPs showed up-regulation in Cu and Zn exposures. Furthermore, HSPs expression in treated groups tended to be higher than the control group. The tendency of CYP450 and GST mRNA expression was higher for Cr and Cu than for other exposure group. The expression of SP gene was low at Al exposure and other group were measured to be similar to control. These results suggest that heavy metal toxicity in freshwater ecosystem may affect physiological and molecular process. Also, the comprehensive gene expression in the aquatic midge Chironomus riparius give useful information to potential molecular biomarkers for assessing heavy metal toxicity.

Tissue Distribution of HuR Protein in Crohn's Disease and IBD Experimental Model (염증성 장질환 모델 및 크론병 환자에서의 점막상피 HuR 단백질의 변화 분석)

  • Choi, Hye Jin;Park, Jae-Hong;Park, Jiyeon;Kim, Juil;Park, Seong-Hwan;Oh, Chang Gyu;Do, Kee Hun;Song, Bo Gyoung;Lee, Seung Joon;Moon, Yuseok
    • Journal of Life Science
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    • v.24 no.12
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    • pp.1339-1344
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    • 2014
  • Inflammatory bowel disease is an immune disorder associated with chronic mucosal inflammation and severe ulceration in the gastrointestinal tract. Antibodies against proinflammatory cytokines, including TNF${\alpha}$, are currently used as promising therapeutic agents against the disease. Stabilization of the transcript is a crucial post-transcriptional process in the expression of proinflammatory cytokines. In the present study, we assessed the expression and histological distribution of the HuR protein, an important transcript stabilizer, in tissues from experimental animals and patients with Crohn's disease. The total and cytosolic levels of the HuR protein were enhanced in the intestinal epithelia from dextran sodium sulfate (DSS)-treated mice compared to those in control tissues from normal mice. Moreover, the expression of HuR was very high only in the mucosal and glandular epithelium, and the relative localization of the protein was sequestered in the lower parts of the villus during the DSS insult. The expression of HuR was significantly higher in mucosal lesions than in normal-looking areas. Consistent with the data from the animal model, the expression of HuR was confined to the mucosal and glandular epithelium. These results suggest that HuR may contribute to the post-transcriptional regulation of proinflammatory genes during early mucosal insults. More mechanistic investigations are warranted to determine the potential use of HuR as a predictive biomarker or a promising target against IBD.

Effect of Trace Metal on Accumulation and Physiological Response of the Polychaete, Perinereis nuntia (미량금속 노출에 따른 갯지렁이(Perinereis nuntia)의 미량금속 축적 및 생리학적인 반응)

  • Won, Eun-Ji;Ra, Kong-Tae;Hong, Seong-Jin;Kim, Kyung-Tae;Lee, Jae-Seong;Shin, Kyung-Hoon
    • Journal of the Korean Society for Marine Environment & Energy
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    • v.13 no.4
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    • pp.288-295
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    • 2010
  • Metal exposure experiments using polychaete (Perinereis nuntia) as a bio-indicator of trace metals contamination were conducted to evaluate the bioaccumulation and the biomarkers responses such as metallothionein-like protein (MTLPs) and glutathione S-transferase (GST) which was simultaneously exposed to Cadmium (Cd) and Copper (Cu). Cu and Cd concentrations in polychaete were enhanced with increasing exposure time and their concentrations of aqueous medium. Initial accumulation of Cd was higher than that of Cu. Our results showed that the bioaccumulation of Cu and Cd were prohibited, especially at higher Cu levels, suggesting the different cellular uptake mechanisms when Cu and Cd are co-exist. Net accumulation rate of Cu was declined with exposure time but it did not show any significant change for Cd. Although the highest MTLPs concentration was observed at 6 hr of exposure time, it did not show any significant change related to exposure times and metals concentrations. An increase of GST activity tended to increase as a function of exposure time and metals concentrations. And GST activities in P. nuntia have similar tendency with bioconcentration factors in high concentration of Cu (treatment group IV) at post 24 h of exposure. Our results provide new information of the bioaccumulation and biomarker responses to understand the effects of co-existing contaminants (Cu and Cd) using polychaete. Further studies are required to elucidate the bioaccumulation and biomarkers responses for various contaminants.

Bioaccumulation of Heavy Metals in Ruditapes philippinarum (바지락 (Ruditapes philippinarum) 의 중금속 축적에 관한 연구)

  • Lee, Yong-Seok;Jo, Yong-Hun;Byun, In-Sun;Kang, Se-Won;Cho, Eun-Mi;Han, Yeon-Soo;Choi, Sang-Haeng;Park, Hong-Seog;Kho, Weon-Gyu;Ahn, In-Young;Jeong, Kye-Heon
    • The Korean Journal of Malacology
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    • v.22 no.2
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    • pp.157-165
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    • 2006
  • The present study was conducted to confirm that a bivalve Ruditapes philippinarum can be used as a biomarker for the monitoring of the heavy metal pollution in the silt of the marine environment. The clams were collected from the silt of Cheonsu-bay, Buheung-ri, and Tan-island of the West Sea, Korea. To observe the normal structures of the target organs (hepatopancreas and gill), they were dissected out for the immunohistochemical study and the electron microscopy with TEM, SEM, and SEM-EDS. The immunohistochemical study showed that the interdiverticular connective tissues of the hepatopancreas, and the outer epithelium of the gill lamellae was strongly reacted to anti-metallothionein (MT), indicating the presence of MT, a metal-binding protein, involved in metal detoxifying process. According to the examinations under the TEM, the epithelial cells of the hepatopancreas of the clams collected from polluted area (Tan-island) showed certain changes such as swollen rER, swollen nuclear envelope and inclusion bodies in the nulcei. In the SEM-EDS analysis, tissue of the hepatopancreas showed relatively higher concentration of S, Zn, and Cd. These elements are supposed to be concerning with the MT-reaction in the hepatopancreas. Considering that the coastal bivalve R. philippinarum showed immediate subcellular responses to heavy metal pollution in the overall experiments conducted, this species might act as one of efficient biomarkers for the heavy metal contamination in the marine environment.

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Increased Antioxidative Activities against Oxidative Stress in Saccharomyces cerevisiae KNU5377 (산화 스트레스 대한 Saccharomyces cerevisiae KNU5377의 항산화 활성의 증가)

  • Kim, Il-Sup;Yun, Hae-Sun;Yang, Ji-Young;Lee, Oh-Seok;Park, Heui-Dong;Jin, Ing-Nyol;Yoon, Ho-Sung
    • Journal of Life Science
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    • v.19 no.4
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    • pp.429-435
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    • 2009
  • Oxidative stress is a consequence of an imbalance of the defense system against cellular damage generated by reactive oxygen species (ROSs) such as superoxide anions (menadione; MD). Most organisms have evolved a variety of defense systems to protect cells from adverse conditions. In order to evaluate stress tolerance against oxidative stress generating MD, comparative analyses of antioxidant capacity, or free radical scavenger ability, were performed between S. cerevisiae KNU5377 (KNU5377) and three wild-type S. cerevisiae strains. In a medium containing 0.4 mM MD, the KNU5377 strain showed higher cell viability and antioxidant ability, and contained higher levels of trehalose, superoxide dismutase, thioredoxin system, glucose-6-phosphate dehydrogenase, and some heat shock proteins. The KNU5377 strain also produced a lower level of oxidative stress biomarker than the other three yeast strains. These results indicate that S. cerevisiae KNU5377 has a higher level of tolerance to oxidative stress due to the increased expression of cell rescue proteins and molecules, thus alleviating cellular damage more efficiently than other S. cerevisiae strains.

MiR-204 acts as a potential therapeutic target in acute myeloid leukemia by increasing BIRC6-mediated apoptosis

  • Wang, Zhiguo;Luo, Hong;Fang, Zehui;Fan, Yanling;Liu, Xiaojuan;Zhang, Yujing;Rui, Shuping;Chen, Yafeng;Hong, Luojia;Gao, Jincheng;Zhang, Mei
    • BMB Reports
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    • v.51 no.9
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    • pp.444-449
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    • 2018
  • Acute myeloid leukemia (AML) is one of the most common hematological malignancies all around the world. MicroRNAs have been determined to contribute various cancers initiation and progression, including AML. Although microRNA-204 (miR-204) exerts anti-tumor effects in several kinds of cancers, its function in AML remains unknown. In the present study, we assessed miR-204 expression in AML blood samples and cell lines. We also investigated the effects of miR-204 on cellular function of AML cells and the underlying mechanisms of the action of miR-204. Our results showed that miR-204 expression was significantly downregulated in AML tissues and cell lines. In addition, overexpression of miR-204 induced growth inhibition and apoptosis in AML cells, including AML5, HL-60, Kasumi-1 and U937 cells. Cell cycle analysis further confirmed an augmentation in theapoptotic subG1 population by miR-204 overexpression. Mechanistically, baculoviral inhibition of apoptosis protein repeat containing 6 (BIRC6) was identified as a direct target of miR-204. Enforcing miR-204 expression increased the luciferase activity and expression of BIRC6, as well as p53 and Bax expression. Moreover, restoration of BIRC6 reversed the pro-apoptotic effects of miR-204 overexpression in AML cells. Taken together, this study demonstrates that miR-204 causes AML cell apoptosis by targeting BIRC6, suggesting miR-204 may play an anti-carcinogenic role in AML and function as a novel biomarker and therapeutic target for the treatment of this disease.

Cloning and Activity Analysis of the FosB Promoter Region from Human Genomic DNA (사람 핵DNA로부터 FosB 유전자 프로모터 클로닝 및 활성도 분석)

  • Na, Han-Heom;Kang, Yoonsung;Kim, Keun-Cheol
    • Journal of Life Science
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    • v.27 no.8
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    • pp.857-863
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    • 2017
  • The FBJ murine osteosarcoma viral oncogene homolog B (FosB) gene is located at chromosome 19, and encodes 43 Kda protein. Functionally, the FosB gene is important for differentiation, development, and pathogenesis. Furthermore, the FosB gene is suggested as possible biomarker for tracing disease prognosis. In this study, we constructed plasmid containing a FosB promoter region and evaluate its promoter activity. We analyzed the putative promoter region in FosB genomic DNA using bioinformatics program, and we found important regulatory elements in 1 Kb upstream from transcription start site (TSS). Therefore, we performed polymerase chain reaction (PCR) amplification on region from-1,555 upstream to +73 of the FosB genomic DNA, and PCR product was inserted into TA vector to create the $TA-1^{st}FosBp$ plasmid. We then prepared the primer sets, which contain a restriction enzyme site for Kpn1 and Nhe1, in order to reinsert into the TA vector to prepare $TA-2^{nd}FosBp$ plasmid. It was finally subcloned into pGL3-luc vector after enzyme cutting. To evaluate whether the cloned plasmid is useful in cell based experiment, we performed luciferase assay with pGL3-FosBp-luctransfection. FosB promoter activity was increased compared to empty vector, and this activity was significantly increased by treatment of doxorubicin and taxol. We obtained consistent data on regulation of FosB gene expression after anticancer drug treatment using Western blot analysis. The results suggest that promoter cloning of the human FosB gene is very useful for studying gene expression and analyzing biomarkers.

A Descriptive Study of Gall Stone Patient's Dietary Habits and Nutritional Status (담석증 환자의 식생활양식 및 영양 실태 조사)

  • Kim, Eun-Jung;Lee, Young-Mee
    • Korean Journal of Community Nutrition
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    • v.12 no.6
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    • pp.826-837
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    • 2007
  • Gallstone composition has changed over the past decades in the Korean with a prominent increase in the prevalence of cholesterol gallstone. This trend is possibly due to the westernization of dietary habits. The purpose of this study was descriptive of GB patient's health related eating behaviors and nutrient consumption patterns. One hundred and six gallstone patients who have had cholecystitis surgery enrolled in this study. Anthropomertic indices, such as height, weight, waist circumference, and hip circumference were measured and calculated BMI and WHR. As the biomarker, TC, TG, LDL-C, HDL-C, FBG level and SBP/DBP were measured and analyzed the relationship with GB stone formation. The structured checklist of health related eating behavior and the semiquantitative food frequency questionnaire after pre-test was used in the face to face interview study. The mean age was $54.9{\pm}13.3$ and gallstone disease was more frequent in the fifties and sixties. Mean BMI was $24.3{\pm}2.8$ in males, and $23.4{\pm}3.9$ in females, the average of waist circumference was $91.1{\pm}7.0cm$ in males and females were $85.4{\pm}9.6cm$. The WHR of men and women was $0.93{\pm}0.0,\;0.90{\pm}0.1$, respectively. The obesity and overweight trend was observed in gallstone patients. The mean blood sugar was researched at $114.5{\pm}35.0mg/dL$. And the ratio of both symptoms DM and gallstone was 26.4%. The rate of dramatic eating repast was significantly higher in the females (p<0.01). Males tended to more frequently consume meat, of high fat content meats and greasy food consumption when eating out. The CPF ratio was 57:16.5:26.5. There was a significant positive correlation between WC and energy (r=0.252, p<0.05), carbohydrate (r=0.255, p<0.05) and niacin (r=0.227, p<0.05). In addition, carbohydrate were significantly correlatied with TC (r=0.230, p<0.05). BMI appeared positive in correlation of protein (r=0.201, p<0.05) and fat (r=0.205, p<0.05). These findings provide a little association that dietary habits are related with cholesterol gallstone formation.

Identifying Genes Related with Radiation Resistance in Oral Squamous Cell Carcinoma Cell Lines (두경부 편평상피세포암주에서 방사선 내성 관련 유전인자의 확인)

  • Lee, Sei-Young;Seol, Jeong-Hun;Park, Haeng-Ran;Cho, Nam-Hoon;Choi, Yoon-Pyo;Rha, Sun-Young;Baek, Seung-Jae;Hwang, Jun-Yeon;Kim, Se-Heon
    • Korean Journal of Head & Neck Oncology
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    • v.27 no.2
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    • pp.190-197
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    • 2011
  • Background and Objective : Radiation resistance(RR) is one of main determinants of treatment outcome in oral squamous cell carcinoma(OSCC), but accurate prediction of RR is difficult. We aim to establish RR OSCC cell lines and identify genes related with RR by a measurement of altered gene expression after inducing RR. Material and Methods : OSCC cell lines, SCC15, SCC25 and QLL1, were treated with 2Gy radiation per session, and parts of them were alive in finally accumulated dosage of 60Gy through 30 times repetition of radiotherapy for inducing RR cell lines. We compared results of cDNA array and proteomics in non-radiated cell lines and RR cell lines to detect changes of gene expression. Western blot was used for the validation of results. Results : cDNA array revealed 265 commonly up-regulated genes and 268 commonly down-regulated genes in 3 RR cell lines comparing their non-radiated counterpart. Among them, 30 cancer related genes were obtained. Proteomics showed 51 commonly altered protein expressions in 3 RR cell lines and 18 cancer related proteins were obtained. Among the detected genes, we found NM23-H1 and PA2G4 were over-expressed in both cDNA array and proteomics. Western blot showed increased expression of NME1 in RR cell lines but not in PA2G4. Conclusion: We concluded that NM23-H1 may be a candidate of RR related gene and over-expression of NM23-H1 could be a biomarker to predict RR in OSCC.