• Title/Summary/Keyword: N25

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The Influence of the N-Terminal Region of Antimicrobial Peptide Pleurocidin on Fungal Apoptosis

  • Choi, Hyemin;Lee, Dong Gun
    • Journal of Microbiology and Biotechnology
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    • v.23 no.10
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    • pp.1386-1394
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    • 2013
  • In our previous study, the 25-mer antimicrobial peptide pleurocidin (Ple) had been thought to induce apoptosis in Candida albicans. This study demonstrated that reactive oxygen species (ROS) production was a major cause of Ple-induced apoptosis. Four truncated analogs were synthesized to understand the functional roles in the N- and C-terminal regions of Ple on the apoptosis. Ple, Ple (4-25), Ple (1-22), and Ple (1-19) produced ROS, including hydroxyl radicals, on the order of [Ple > Ple (1-22) > Ple (4-25) > Ple (1-19)], whereas Ple (7-25) did not induce any ROS production. The results suggested that the N-terminal deletion affected the ROS-inducing activities much more than that of the C-terminal deletion, and net hydrophobicity [Ple > Ple (1-22) > Ple (4-25) > Ple (1-19) > Ple (7-25)] was related to ROS generation rather than other primary factors like net charge. Hence, we focused on the N-terminal-truncated peptides, Ple (4-25) and Ple (7-25), and examined other apoptotic features, including mitochondrial membrane depolarization, caspase activation, phosphatidylserine externalization, and DNA and nuclear fragmentation. The results also confirmed the disappearance of apoptotic activity of Ple (7-25) by the truncation of the N-terminal region (1-6) and the specific activity patterns between Ple and analogs. In conclusion, the N-terminal region of Ple played an important role in apoptosis.

A Novel Suberoylanilide Hydroxamic Acid Histone Deacetylase Inhibitor Derivative, N25, Exhibiting Improved Antitumor Activity in both Human U251 and H460 Cells

  • Zhang, Song;Huang, Wei-Bin;Wu, Li;Wang, Lai-You;Ye, Lian-Bao;Feng, Bing-Hong
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.10
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    • pp.4331-4338
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    • 2014
  • $N^1$-(2, 5-dimethoxyphenyl)-$N^8$-hydroxyoctanediamide (N25) is a novel SAHA cap derivative of HDACi, with a patent (No. CN 103159646). This invention is a hydroxamic acid compound with a structural formula of $RNHCO(CH_2)6CONHOH$ (wherein R=2, 5dimethoxyaniline), a pharmaceutically acceptable salt which is soluble. In the present study, we investigated the effects of N25 with regard to drug distribution and molecular docking, and anti-proliferation, apoptosis, cell cycling, and $LD_{50}$. First, we designed a molecular approach for modeling selected SAHA derivatives based on available structural information regarding human HDAC8 in complex with SAHA (PDB code 1T69). N25 was found to be stabilized by direct interaction with the HDAC8. Anti-proliferative activity was observed in human glioma U251, U87, T98G cells and human lung cancer H460, A549, H1299 cells at moderate concentrations ($0.5-30{\mu}M$). Compared with SAHA, N25 displayed an increased antitumor activity in U251 and H460 cells. We further analyzed cell death mechanisms activated by N25 in U251 and H460 cells. N25 significantly increased acetylation of Histone 3 and inhibited HDAC4. On RT-PCR analysis, N25 increased the mRNA levels of p21, however, decreased the levels of p53. These resulted in promotion of apoptosis, inducing G0/G1 arrest in U251 cells and G2/M arrest in H460 cells in a time-dependent and dose-dependent manner. In addition, N25 was able to distribute to brain tissue through the blood-brain barrier of mice ($LD_{50}$: 240.840mg/kg). In conclusion, our findings demonstrate that N25 will provide an invaluable tool to investigate the molecular mechanism with potential chemotherapeutic value in several malignancies, especially human glioma.

THE EFFECT OF $1,25-(OH)_2D_3$ ON THE PROLIFERATION AND ALKALINE PHOSPHATASE ACTIVITY OF HUMAN PERIODONTAL LIGAMENT CELLS (치주인대세포의 증식 및 세포활성에 미치는 $1,25-(OH)_2D_3$의 영향에 관한 연구)

  • Kook, Yoon-Ah;Kim, Sang-Cheol;Kim, Hyung-Ryong
    • The korean journal of orthodontics
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    • v.25 no.3 s.50
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    • pp.333-339
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    • 1995
  • The hormonally active vitamin D metabolite, 1,25-dihydroxy vitamin $D_3[1.25-(OH)_2D_3]$ is one of the several humoral factors that may regulate osteoblast differentiation. The purpose of this study was to evaluate the effects of $1,25-(OH)_2D_3$ on the PDL cells. Human PDL cells were prepared from the first premolar tooth extracted for the orthodontic treatment and they were incubated in the environment of $37^{\circ}C,\;5\%\;CO_2\;and\;95\%$ humidity. $[{^3}H]$-thymidine incorporation as a measure of proliferation potential and alkaline phosphatase activity were evaluated at 10nM, 100nM $1,25-(OH)_2D_3$. The observed results were as follows. 1. $1,25-(OH)_2D_3$ was significantly enhanced $[{^3}H]$-thymidine incorporation at 100nM, But did not affect by 10nM. 2. $1,25-(OH)_2D_3$ was significantly increased alkaline phosphatase activity at 1 day and 6 days in a dose-dependent manner.

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Evaluation on Usefulness of BMI Application to Urological CT Examination (비뇨기계 CT 검사 시 체질량 지수 적용의 유용성 평가)

  • Kim, Hyeon-jin;Im, In-chul
    • Journal of the Korean Society of Radiology
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    • v.12 no.2
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    • pp.185-191
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    • 2018
  • The purpose of this study was to evaluate the usefulness of BMI application to Urography CT by applying different tube voltages in accordance with body mass index. Group A (n = 38) with body mass index of lower than 25 was examined with tube voltage of 100 kVp while Group B (n = 45) with a BMI of 25 and higher was examined with tube voltage of 120 kVp. C group (n = 37) with body mass index (BMI) of lower than 25 was examined with tube voltage of 120kVp. Although the difference in average dose between group A (100 kVp) and group C (120 kVp) with low body mass index (BMI) of lower than 25 was $214.8mGy{\cdot}cm$, there was no significant difference in qualitative evaluation and, compared with patient group with body mass index of 25 and higher, results obtained were rather good. Therefore, this study verified that the tube voltage of lower than 100 kVp does not have adverse effect on the quality of image for patients with body mass index (BMI) of lower than 25.

Influence of the Hydrophobic Amino Acids in the N- and C-Terminal Regions of Pleurocidin on Antifungal Activity

  • Lee, June-Young;Lee, Dong-Gun
    • Journal of Microbiology and Biotechnology
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    • v.20 no.8
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    • pp.1192-1195
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    • 2010
  • To investigate the influence of the N- or C-terminal regions of pleurocidin (Ple) peptide on antifungal activity, four analogs partially truncated in the N- or C-terminal regions were designed and synthesized. Circular dichroism (CD) spectroscopy demonstrated that all the analogs maintained an alpha-helical structure. The antifungal susceptibility testing also showed that the analogs exhibited antifungal activities against human fungal pathogens, without hemolytic effects against human erythrocytes. The result further indicated that the analogs had discrepant antifungal activities [Ple>Ple (1-22)>Ple (4-25)>Ple (1- 19)>Ple (7-25)] and that N-terminal deletion affected the activities much more than C-terminal deletion. Hydrophobicity [Ple>Ple (1-22)>Ple (4-25)>Ple (1-19)> Ple (7-25)] was thought to have been one of the consistent factors that influenced these activity patterns, rather than the other primary factors like the helicity [Ple>Ple (4-25) >Ple (1-22)>Ple (1-19)>Ple (7-25)] or the net charge [Ple=Ple (4-25)=Ple (7-25)>Ple (1-22)=Ple (1-19)] of the peptides. In conclusion, the hydrophobic amino acids in the N-terminal region of Ple is more crucial for antifungal activity than those in the C-terminal region.

The Binding Properties of Glycosylated and Non- Glycosylated Tim-3 Molecules on $CD4^+CD25^+$T Cells

  • Lee, Mi-Jin;Heo, Yoo-Mi;Hong, Seung-Ho;Kim, Kyong-Min;Park, Sun
    • IMMUNE NETWORK
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    • v.9 no.2
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    • pp.58-63
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    • 2009
  • Background: T cell immunoglobulin and mucin domain containing 3 protein (Tim-3) expressed on terminally differentiated Th1 cells plays a suppressive role in Th1-mediated immune responses. Recently, it has been shown that N-glycosylation affects the binding activity of the Tim-3-Ig fusion protein to its ligand, galectin-9, but the binding properties of non-glycosylated Tim-3 on $CD4^+CD25^+$T cells has not been fully examined. In this study, we produced recombinant Tim-3-Ig fusion proteins in different cellular sources and its N-glycosylation mutant forms to evaluate their binding activities to $CD4^+CD25^+$T cells. Methods: We isolated and cloned Tim-3 cDNA from BALB/C mouse splenocytes. Then, we constructed a mammalian expression vector and a prokaryotic expression vector for the Tim-3-Ig fusion protein. Using a site directed mutagenesis method, plasmid vectors for Tim-3-Ig N-glycosylation mutant expression were produced. The recombinant protein was purified by protein A sepharose column chromatography. The binding activity of Tim-3-Ig fusion protein to $CD4^+CD25^+$T cells was analyzed using flow cytometry. Results: We found that the nonglycosylated Tim-3-Ig fusion proteins expressed in bacteria bound to $CD4^+CD25^+$T cells similarly to the glycosylated Tim-3-Ig protein produced in CHO cells. Further, three N-glycosylation mutant forms (N53Q, N100Q, N53/100Q) of Tim-3-Ig showed similar binding activities to those of wild type glycosylated Tim-3-Ig. Conclusion: Our results suggest that N-glycosylation of Tim-3 may not affect its binding activity to ligands expressed on $CD4^+CD25^+$T cells.

Chemical Analysis of Cuticular Hydrocarbons in Apis mellifera L. and Apis cerana F. (동양종과 서양종 꿀벌의 표피탄화수소 성분 분석)

  • 이창주;신경우;박승찬;심재한
    • Korean journal of applied entomology
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    • v.42 no.1
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    • pp.9-13
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    • 2003
  • Cuticular hydrocarbons of antenna, legs and wings from two species of honeybee worker of Apis mellifera L. and Apis cerana F. can be analyzed directly with gas chromatograph and GC/MS without solvent extraction. The saturated hydrocarbons identified in selected part of both species were nC22, nC23, nC25-nC3O, nC32 and nC34 except nC24. Two saturated hydrocarbons, nC26 (23.0-42.6%) and nC28 (16.8-54.8%), were major compounds in both species and others were minor compounds. A. mellifera can be distinguished from A. cerana F. by having higher proportion of nC30, nC32 and nC34 by having lower proportion of nC25 from three selected part of both species.

N-IDEALS OF SUBTRACTION ALGEBRAS

  • Jun, Young-Bae;Kavikumar, Jacob;So, Keum-Sook
    • Communications of the Korean Mathematical Society
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    • v.25 no.2
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    • pp.173-184
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    • 2010
  • Using $\cal{N}$-structures, the notion of an $\cal{N}$-ideal in a subtraction algebra is introduced. Characterizations of an $\cal{N}$-ideal are discussed. Conditions for an $\cal{N}$-structure to be an $\cal{N}$-ideal are provided. The description of a created $\cal{N}$-ideal is established.