• Title/Summary/Keyword: expression profile

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The Effect of Reaction Temperature and Volume in the Sonolysis of 1,4-Dioxane (1,4-Dioxane의 초음파 처리시 반응 온도와 부칙의 영향)

  • Son, Hyun-Seok;Choi, Seok-Bong;Eakalak, Khan;Zoh, Kyung-Duk
    • Journal of Korean Society of Environmental Engineers
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    • v.27 no.10
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    • pp.1114-1122
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    • 2005
  • This research investigates the effects of sonication mode, reaction temperature and volume on sonolysis of 1,4-dioxane in order to increase the degradation efficiency and kinetics. The degradation efficiency in case with pulse mode was about 10 % higher than that in case with continuous mode. The degradation profiles in both cases, which were performed without the control of reaction temperature and in 1000 mL, were composed of three steps. However, 1,4-D was mainly degraded in the initiation step as the first portion and the acceleration step as the second portion. The initial step agreed with zero-order expression well, while the acceleration step could be fitted with pseudo 1st-order expression. The kinetic model in case with $5^{\circ}C$ and 300 mL conformed to pseudo 1st-order, while that in cases with $10^{\circ}C$ to $40^{\circ}C$ agreed with zero-order expression. The degradation efficiency and profile of 1,4-D in the experiment with $20^{\circ}C$ and 300 mL was higher and simpler than that in case with $20^{\circ}C$ and 1000 mL. The reaction temperature and volume influence bubble intensity, which was produced in sonication. The increase of bubble intensity induced to augment the production of OH radical in sonication.

A truncated form of human alpha 1-acid glycoprotein is useful as a molecular tool for insect glycobiology

  • Morokuma, Daisuke;Hino, Masato;Tsuchioka, Miho;Masuda, Akitsu;Mon, Hiroaki;Fujiyama, Kazuhito;Kajiura, Hiroyuki;Kusakabe, Takahiro;Lee, Jae Man
    • International Journal of Industrial Entomology and Biomaterials
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    • v.36 no.1
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    • pp.15-24
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    • 2018
  • N-glycosylation is an important posttranslational modification that results in a variety of biological activities, structural stability, and protein-protein interactions. There are still many mysteries in the structure and function of N-glycans, and detailed elucidation is necessary. Baculovirus expression system (BES) is widely used to produce recombinant glycoproteins, but it is not suitable for clinical use due to differences in N-glycan structure between insects and mammals. It is necessary to develop adequate model glycoproteins for analysis to efficiently alter the insect-type N-glycosylation pathway to human type. The previous research shows the recombinant alpha 1-acid glycoprotein (${\alpha}1AGP$) secreted from silkworm cultured cells or larvae is highly glycosylated and expected to be an excellent research candidate for the glycoprotein analysis expressed by BES. Therefore, we improved the ${\alpha}1AGP$ to be a better model for studying glycosylation. The modified ${\alpha}1AGP$ (${\alpha}1AGP{\Delta}$) recombinant protein was successfully expressed and purified by using BES, however, the expression level in silkworm cultured cells and larvae were lower than that of the ${\alpha}1AGP$. Subsequently, we confirmed the detailed profile of N-glycan on the ${\alpha}1AGP{\Delta}$ by LS/MS analysis the N-glycan structure at each glycosylation site. These results indicated that the recombinant ${\alpha}1AGP{\Delta}$ could be usable as a better model glycoprotein of N-glycosylation research in BES.

Combined Expression of Metastasis Related Markers Naa10p, SNCG and PRL-3 and its Prognostic Value in Breast Cancer Patients

  • Min, Li;Ma, Ruo-Lan;Yuan, Hua;Liu, Cai-Yun;Dong, Bing;Zhang, Cheng;Zeng, Yan;Wang, Li;Guo, Jian-Ping;Qu, Li-Ke;Shou, Cheng-Chao
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.7
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    • pp.2819-2826
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    • 2015
  • Combinations of multiple biomarkers representing distinct aspects of metastasis may have better prognostic value for breast cancer patients, especially those in late stages. In this study, we evaluated the protein levels of N-${\alpha}$-acetyltransferase 10 protein (Naa10p), synuclein-${\gamma}$ (SNCG), and phosphatase of regenerating liver-3 (PRL-3) in 365 patients with breast cancer by immunohistochemistry. Distinct prognostic subgroups of breast cancer were identified by combination of the three biomarkers. The Naa10p+SNCG-PRL-3-subgroup showed best prognosis with a median distant metastasis-free survival (DMFS) of 140 months, while the Naa10p-SNCG+PRL-3+subgroup had the worst prognosis with a median DMFS of 60.5 months. Multivariate analysis indicated Naa10p, SNCG, PRL-3, and the TNM classification were all independent prognostic factors for both DMFS and overall survival (OS). The three biomarker combination of Naa10p, SNCG and PRL-3 performed better in patients with lymph node metastasis, especially those with more advanced tumors than other subgroups. In conclusion, the combined expression profile of Naa10p, SNCG and PRL-3, alone or in combination with the TNM classification system, may provide a precise estimate of prognosis of breast cancer patients.

cDNA microarray analysis of viral hemorrhagic septicemia infected olive flounder, Paralichthys olivaceus: immune gene expression at different water temperature (바이러스성 출혈성 패혈증에 감염된 넙치의 cDNA microarray 분석 : 수온에 따른 면역 유전자 발현의 차이)

  • Kim, Jin-Ung;Jung, Sung-Ju
    • Journal of fish pathology
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    • v.27 no.1
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    • pp.1-9
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    • 2014
  • The olive flounder, Paralichthys olivaceus is susceptible to viral hemorrhagic septicaemia virus (VHSV) at $15^{\circ}C$ but no mortality at $20^{\circ}C$ even though the virus can grow well in vitro at $20^{\circ}C$. Thus, we designed an experiment to know immune response of olive flounder against VHSV when the host reared at $15^{\circ}C$ or $20^{\circ}C$. cDNA microarray analysis was performed to compare the gene expression patterns of the kidney cells between the host reared at $15^{\circ}C$ or $20^{\circ}C$. The expression of MHC class I, IL-8, myeloperoxidae and endonuclease G-like having function for the antigen presentation and chemokine-factor were up-regulted both the $15^{\circ}C$ and $20^{\circ}C$ during VHSV infection. MHC class II gene existing on antigen-presenting cells and B cell lymphocytes, immunoglobulin (Ig) genes and phagocytosis related genes were down-regulated at $15^{\circ}C$ but highly expressed at $20^{\circ}C$. It can be thought that innate immune related antigen presentation by MHC class I and phagocytosis reaction against VHSV are efficiently occur both the temperature but macrophage or B cell related antigen presentation via MHC class II fails to induce downstream immune reactions (adaptive immunity) to make antibody, and it can be one of the reason that causes high mortality only at $15^{\circ}C$.

Identification of Genes Involved in Primordial-primary Follicle Transition by Suppression Subtractive Hybridization

  • Park, Chang-Eun;Yoon, Se-Jin;Jeon, Eun-Hyun;Kim, Young-Hoon;Lee, Sook-Hwan;Lee, Kyung-Ah
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.98-98
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    • 2002
  • Recruitment of primordial follicles(PMF) is crucial for female fertility. however, factors and mechanisms that regulate this process is poorly understood. The present study was conducted to obtain an inclusive view of the gene expression and to identify novel factors and their pathways of regulating PMF arrest and/or growth initiation. Ovaries from one-day neonatal(consists of oocyte and PMF) and five-day old(consists of PMF and primary follicles, PRIF) mice were collected, either total RNA or mRNA was isolated, and suppression subtractive hybridization(SSH) was used to isolate and clone genes that differentially expressed in day 1 and day 5 ovaries. Confirmation that some of these genes are differentially expressed in PMF and/or in PRIF was accomplished by using laser captured microdissection(LCM), RT-PCR. in situ hybridization(ISH) and/or immunohistochemistry(IHC). In toto, 357 clones were sequenced and analyzed by BLAST and RIKEN program. Sequences of 330 clones significantly matched database entries while 27 clones were novel. Forty-two and 47 different genes were identified as differentially expressed in day 1 and day 5 ovaries, respectively, while 7 genes were expressed in both stages of ovaries. Day 5-subtracted library included several genes known as markers far growing follicles, such as ZP2, MATER, and fetuin. Among the genes with assigned functions, 23.8% was associated with cell cycle/apoptosis regulation, 7.1% with cellular structure, 11.9% with metabolism, 26.2% with signal transduction, and 31.0% with gene/protein expression in day 1; while 10.6%, 17.0%, 23.5%, 25.5%, and 23.4% in day 5, respectively. Genes such as GDF-8, Lats2, Septin2, and Weel were the highly expressed genes in PMF, while HSP84, Laminin2, MATER, MTi7, PTP, and Wrn were highly expressed genes in PRIF. We have successfully discovered list of genes expressed in day 1 and day 5 ovaries and confirmed that some of them are differentially expressed in PMF and/or PRIF. Gene expression profile from the present study would provide insight for the future study on the mechanism(s) involved in primordial-primary follicular transition. This work was Supported by Korean Health 21 RND Project, Ministry of Health and Welfare, Korea (01-PJ10-PG6-01GN13-0002).

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Circulating Cell-free miRNA Expression and its Association with Clinicopathologic Features in Inflammatory and Non-Inflammatory Breast Cancer

  • Hamdi, K;Blancato, J;Goerlitz, D;Islam, MD;Neili, B;Abidi, A;Gat, A;Ayed, F Ben;Chivi, S;Loffredo, CA;Jillson, I;Elgaaied, A Benammar;Marrakchi, R
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.4
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    • pp.1801-1810
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    • 2016
  • Recent discovery showing the presence of microRNAs (miRNAs) in the circulation sparked interest in their use as potential biomarkers. Our previous studies showed the diagnostic potential of miR-451 as a serological marker for inflammatory breast cancer (IBC), miR-337-5p and miR-30b for non-inflammatory breast cancer (non-IBC). The aim of this study is to investigate the prognostic values of circulating miRNAs by comparing the amounts of 12 circulating miRNAs in the serum of IBC and non-IBC from Tunisian breast cancer patients, and by determinating whether correlated pairs of miRNAs could provide useful information in the diagnosis of IBC and non-IBC patients. TaqMan qPCR was performed to detect circulating expression of miRNAs in serum of 20 IBC, 20 non-IBC and 20 healthy controls. Nonparametric rank Spearman rho correlation coefficient was used to examine the prognostic value of miRNAs and to assess the correlation profile between miRNAs expression. Further, a large number of miRNAs were highly correlated (rho>0.5) in both patients groups and controls. Also, the correlations profiles were different between IBC, non-IBC and healthy controls indicating important changes in molecular pathways in cancer cells. Our results showed that miR-335 was significantly overexpressed in premenopausal non-IBC patients; miR-24 was significantly overexpressed in non-IBC postmenopausal patients. Patients with previous parity had higher serum of miR-342-5p levels than those without. Furthermore, patients with HER2+ IBC present lower serum levels of miR-15a than patients with HER2-disease. Together, these results underline the potential of miRNAs to function as diagnostic and prognostic markers for IBC and non-IBC, with links to the menopausal state, Her2 status and parity.

Effect of Red Yeast (Monascus purpureus) Rice Supplemented Diet on Lipid Profiles and Antioxidant Activity in Hypercholesterolemic Rats (홍국(Monascus purpureus)쌀을 첨가한 고콜레스테롤 식이가 흰쥐의 항산화 활성에 미치는 영향)

  • Kwon, Chong Suk
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.1
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    • pp.16-23
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    • 2014
  • Red yeast rice (RYR) has been known to exhibit various biological effects, including anti-hyperlipidemia, antioxidant, anti-tumor, and anti-inflammtory activities. Oxidative stress is a main risk factor in the development of cardiovascular disease, such as atherosclerosis. Therefore, the aim of this study was to investigate the possible hypolipidemic and antioxidant effect of RYR on rats fed a high-cholesterol diet supplemented with either 0.2%, 1%, or 5% RYR for 4 weeks. We measured lipid profiles in the plasma and liver, antioxidant enzyme activities in plasma and erythrocyte, gene expression of antioxidant enzymes in the liver, and oxidative DNA damage in lymphocytes. The group supplemented with 0.2% RYR had total cholesterol level in plasma decreased by 24%, while the group supplemented with 5% RYR had high-density cholesterol increased by 20% compared to the control. The antioxidant enzyme activities were also affected by RYR supplementation. Total superoxide dismutase activities in plasma significantly decreased by 11% in the 1% RYR group, while these activities in the liver significantly decreased by 16% and 21% in the 1% and 5% supplemented group compared to the control, respectively. Glutathione peroxidase activities in plasma and erythrocytes increased 13% and 48% in the 1% RYR group, respectively. Catalase (CAT) activity in erythrocytes significantly increased by 49% and 68% in the 1% and 5% RYR group compared to the control, respectively. The gene expression of CAT was up-regulated 7.9 fold compared to the control in the 5% RYR supplemented group. These results suggest that RYR can control hyperlipidemia by improving the lipid profile and modulating oxidative stress.

MicroRNA-23a: A Novel Serum Based Diagnostic Biomarker for Lung Adenocarcinoma

  • Lee, Yu-Mi;Cho, Hyun-Jung;Lee, Soo-Young;Yun, Seong-Cheol;Kim, Ji-Hye;Lee, Shin-Yup;Kwon, Sun-Jung;Choi, Eu-Gene;Na, Moon-Jun;Kang, Jae-Ku;Son, Ji-Woong
    • Tuberculosis and Respiratory Diseases
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    • v.71 no.1
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    • pp.8-14
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    • 2011
  • Background: MicroRNAs (miRNAs) have demonstrated their potential as biomarkers for lung cancer diagnosis. In recent years, miRNAs have been found in body fluids such as serum, plasma, urine and saliva. Circulating miRNAs are highly stable and resistant to RNase activity along with, extreme pH and temperatures in serum and plasma. In this study, we investigated serum miRNA profiles that can be used as a diagnostic biomarker of non-small cell lung cancer (NSCLC). Methods: We compared the expression profile of miRNAs in the plasma of patients diagnosed with lung cancer using an miRNA microarray. The data from this assay were validated by quantitative real-time PCR (qRT-PCR). Results: Six miRNAs were overexpressed and three miRNAs were underexpressed in both tissue and serum from squamous cell carcinoma (SCC) patients. Sixteen miRNAs were overexpressed and twenty two miRNAs were underexpressed in both tissue and serum from adenocarcinoma (AC) patients. Of the four miRNAs chosen for qRT-PCR analysis, the expression of miR-23a was consistent with microarray results from AC patients. Receiver operating characteristic (ROC) curve analyses were done and revealed that the level of serum miR-23a was a potential marker for discriminating AC patients from chronic obstructive pulmonary disease (COPD) patients. Conclusion: Although a small number of patients were examined, the results from our study suggest that serum miR-23a can be used in the diagnosis of AC.

SB202190- and SB203580-Sensitive p38 Mitogen-Activated Protein Kinase Positively Regulates Heat Shock- and Amino Acid Analog-Induced Heat Shock Protein Expression

  • Kim, Sun-Hee;Han, Song-Iy;Oh, Su-Young;Seo, Myoung-Suk;Park, Hye-Gyeong;Kang, Ho-Sung
    • Biomedical Science Letters
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    • v.9 no.2
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    • pp.59-65
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    • 2003
  • When cells are exposed to proteotoxic stresses such as heat shock, amino acid analogs, and heavy metals, they increase the synthesis of the heat shock proteins (HSPs) by activating the heat shock transcription factor 1 (HSF1), whose activity is controlled via multiple steps including homotrimerization, nuclear translocation, DNA binding, and hyperphosphorylation. Under unstressed conditions, the HSF1 activity is repressed through its constitutive phosphorylation by glycogen synthase kinase 3$\beta$ (GSK3$\beta$), extracellular regulated kinase 1/2 (ERK1/2), and stress-activated protein kinase/c-Jun N-terminal kinase (SAPK/JNK). However, the protein kinase (s) responsible for HSF1 hyperphosphorylation and activation is not yet identified. In the present study, we observed that profile of p38 mitogen-activated protein kinase (p38MAPK) activation in response to heat shock was very similar to those of HSF1 hyperphosphorylation and nuclear translocation. Therefore, we investigated whether p38MAPK is involved in the heat shock-induced HSF1 activation and HSP expression. Here we show that the p38MAPK inhibitors, SB202190 and SB203580, but not other inhibitors including the MEK1/2 inhibitor PD98059 and the PI3-K inhibitor LY294002 and wortmannin, suppress HSF1 hyperphosphorylation in response to heat shock and L-azetidine 2-carboxylic acid (Azc), but not to heavy metals. Furthermore, heat shock-induced HSF1-DNA binding and HSP72 expression was specifically prevented by the p38MAPK inhibitors, but not by the MEK1/2 inhibitor and the PI3-K inhibitors. These results suggest that SB202190- and SB203580-sensitive p38MAPK may positively regulate HSP gene regulation in response to heat shock and amino acid analogs.

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OmpR Is Essential for Growth and Expression of Virulence-related Genes in the Fish Pathogen Edwardsiella piscicida (어류 병원체 Edwardsiella piscicida의 OmpR은 생육과 병원성과 관련된 유전자의 발현에 필수적)

  • Ray, Durga;Kim, Yeon Ha;Choe, unjeong;Kang, Ho Young
    • Journal of Life Science
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    • v.31 no.1
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    • pp.28-36
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    • 2021
  • Edwardsiella piscicida is a significant cause of hemorrhagic septicemia in fish and gastrointestinal infections in humans. Survival bacteria require specialized mechanisms to adapt to environmental fluctuations. Hence, to understand the mechanism through which E. piscicida senses and responds to environmental osmolarity changes, we determined the protein expression profile and physiological properties under various salinity conditions in this study. The OmpR protein is a part of the Env-ZOmpR two-component system that has been implicated in sensing salt stress in bacteria. However, the physiological role played by this protein in E. piscicida remains to be elucidated. Therefore, in this work, the function of the OmpR protein in response to salt stress was investigated. Phenotypic analysis revealed that, in the mutant, three of the biochemical phenotypes were different from the wild type, including, citrate utilization, hydrogen sulfide, and indole production. Introduction of the plasmid containing the entire ompR gene to the mutant strain returned it to its parental phenotype. The retarded growth rate also partially recovered. Furthermore, in our studies, OmpR was not found to be related to cell motility. Taken together, our results from the mutational analysis, the growth assay, MALDI-TOF MS, qRT-PCR, and the phenotype studies suggest that the OmpR of E. piscicida is implicated in osmoregulation, growth, expression of porins (ETAE_1826), virulence-related genes (EseC, EseD and EvpC), and certain genes of unknown function (ETAE_1540 and ETAE_2706).