Effective suppression of C5a-induced proinflammatory response using anti-human C5a repebody

Hwang, D.E.,Choi, J.M.,Yang, C.S.,Lee, J.j.,Heu, W.,Jo, E.K.,Kim, H.S. Academic Press 2016 Biochemical and biophysical research communication Vol.477 No.4

The strongest anaphylatoxin, C5a, plays a critical role in the proinflammatory responses, causing the pathogenesis of a number of inflammatory diseases including sepsis, asthma, and rheumatoid arthritis. Inhibitors of C5a thus have great potential as therapeutics for various inflammatory disorders. Herein, we present the development of a high-affinity repebody against human C5a (hC5a), which effectively suppresses the proinflammatory response. A repebody scaffold composed of leucine-rich repeat (LRR) modules was previously developed as an alternative protein scaffold. A repebody specifically binding to hC5a was selected through a phage display, and its affinity was increased up to 5 nM using modular engineering. The repebody was shown to effectively inhibit the production of C5a-induced proinflammatory cytokines by human monocytes. To obtain insight into a mode of action by the repebody, we determined its crystal structure in complex with hC5a. A structural analysis revealed that the repebody binds to the D1 and D3 regions of hC5a, overlapping several epitope residues with the hC5a receptor (hC5aR). It is thus likely that the repebody suppresses the hC5a-mediated immune response in monocytes by blocking the binding of hC5a to its receptor. The anti-hC5a repebody can be developed as a potential therapeutic for C5a-involved inflammatory diseases.

Metal(loid) immobilization in soils with biochars pyrolyzed in N₂ and CO₂ environments

Igalavithana, Avanthi Deshani,Yang, Xiao,Zahra, Hilda Rizkia,Tack, Filip M.G.,Tsang, Daniel C.W.,Kwon, Eilhann E.,Ok, Yong Sik Elsevier 2018 Science of the Total Environment Vol.630 No.-

<P><B>Abstract</B></P> <P>Previous studies indicated that using CO<SUB>2</SUB> as a reaction agent in the pyrolysis of biomass led to an enhanced generation of syngas <I>via</I> direct reaction between volatile organic carbons (VOCs) evolved from the thermal degradation of biomass and CO<SUB>2</SUB>. In addition, the physico-chemical properties of biochar in CO<SUB>2</SUB> were modified. In this current study, biochars generated from red pepper stalks in N<SUB>2</SUB> and CO<SUB>2</SUB> (RPS-N and RPS-C, respectively) were tested for their effects on the immobilization of Pb, Cd, Zn, and As in contaminated soils. Soils were incubated for one month with 2.5% of RPS, and two biochars (<I>i.e.</I>, RPS-N and RPS-C) at 25°C. After the incubation period soils were analyzed to determine the amendment effects on the behavior of metal(loid)s. The potential availability and mobility kinetics of metal(loid)s were assessed by single extraction of ammonium acetate and consecutive extraction of calcium chloride, respectively. Sequential extraction was used to further examine potential changes in geochemical fractions of metal(loid)s. The increased soil pH induced by application of the biochars reduced the potentially available Pb, Cd, and Zn, while RPS-C significantly reduced Pb due to the high surface area and aromaticity of RPS-C. However, RPS-C mobilized potentially available As compared to RPS-N due to the increased soil pH. Biochars reduced the mobility kinetics of Pb, Cd, and Zn, and RPS-N effectuated the greatest reduction of As mobility. The RPS-C increased the Fe and Mn oxides, hydroxide, and organically bound Pb, while both biochars and RPS-N increased residual Cd and Zn, and organically bound As, respectively. When considering the two biochars, RPS-C was highly effective for immobilization of Pb in soils, but it had no effect on Cd and Zn and a negative effect on As. In addition, RPS-C significantly increased the total exchangeable cations in soils.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Biochars markedly reduced Cd, Pb and Zn mobility in soils. </LI> <LI> Biochar pyrolyzed in CO<SUB>2</SUB> most effectively immobilized Pb. </LI> <LI> Biochar pyrolyzed in CO<SUB>2</SUB> increases As mobility more. </LI> <LI> More siloxane groups present in biochar pyrolysed in CO<SUB>2</SUB>. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

Development of additive [¹¹C]CO₂ target system in the KOTRON-13 cyclotron and its application for [¹¹C]radiopharmaceutical production

Moon, B.S.,Lee, H.J.,Lee, W.K.,Hur, M.G.,Yang, S.D.,Lee, B.C.,Kim, S.E. North-Holland Physics Pub 2015 Nuclear instruments & methods in physics research. Vol.356 No.-

The KOTRON-13 cyclotron, which was developed in South Korea for the production of medical radioisotopes, has the structural limitation of only one beam-output port, restricting the production of the carbon-11 isotope. In the present study, we investigate the design of a switchable target system and develop an effective carbon-11 target in the KOTRON-13 cyclotron, for combination with the fluorine-18 target. The target system was designed by introducing a sliding-type element between the fluorine-18 and carbon-11 targets, a tailor-made C-11 target and its cooling system. For the efficient production of [<SUP>11</SUP>C]CO<SUB>2</SUB>, the desirable target shape and internal volume were determined by a Stopping and Range of Ions in Matter (SRIM) simulation program, and the target grid was modified to resist the cavity pressure during beam irradiation. We evaluated the [<SUP>11</SUP>C]CO<SUB>2</SUB> production while varying the material and thickness of the target foil, oxygen content of the nitrogen gas, and target loading pressure. Using sliding-type equipment including an additional gate valve and a high vacuum in a beam line, the bi-directional conversion between the fluorine-18 and carbon-11 targets was efficient regarding the accurate beam irradiation on both targets. The optimal [<SUP>11</SUP>C]CO<SUB>2</SUB> production for 30min irradiation at 60μA (86.6+/-1.7GBq in the target at EOB) was observed at a thickness of 19μm with HAVAR® material as a target foil and a target loading pressure of 24bar with nitrogen plus 300ppb of oxygen gas. Additionally, the coolant cavity system in the target grid and target chamber is useful to remove the heat transferred to the target body by the internal convection of water and thereby ensure the stability of the [<SUP>11</SUP>C]CO<SUB>2</SUB> production under a high beam current. In the application of C-11 labeled radiopharmaceuticals such as [<SUP>11</SUP>C]PIB, [<SUP>11</SUP>C]DASB, [<SUP>11</SUP>C]PBR28, [<SUP>11</SUP>C]Methionine and [<SUP>11</SUP>C]Clozapine, the radiochemical yields were shown to be 25-38% (decay corrected) with over 166GBq/μmol of specific activity. Consequently, the additive carbon-11 target system was successfully developed in only one output port of the KOTRON-13 cyclotron and exhibited the stable production of C-11 labeled radiopharmaceuticals.

Gα₁₂ gep oncogene deregulation of p53-responsive microRNAs promotes epithelial–mesenchymal transition of hepatocellular carcinoma

Yang, Y M,Lee, W H,Lee, C G,An, J,Kim, E-S,Kim, S H,Lee, S-K,Lee, C H,Dhanasekaran, D N,Moon, A,Hwang, S,Lee, S J,Park, J-W,Kim, K M,Kim, S G Macmillan Publishers Limited 2015 Oncogene Vol.34 No.22

Hepatocellular carcinoma (HCC) has a poor prognosis owing to aggressive phenotype. Gα<SUB>12</SUB> gep oncogene product couples to G-protein-coupled receptors, whose ligand levels are frequently increased in tumor microenvironments. Here, we report Gα<SUB>12</SUB> overexpression in human HCC and the resultant induction of zinc-finger E-box-binding homeobox 1 (ZEB1) as mediated by microRNA deregulation. Gα<SUB>12</SUB> expression was higher in HCC than surrounding non-tumorous tissue. Transfection of Huh7 cell with an activated mutant of Gα<SUB>12</SUB> (Gα<SUB>12</SUB>QL) deregulated microRNA (miRNA or miR)-200b/a/429, -194-2/192 and -194-1/215 clusters in the miRNome. cDNA microarray analyses disclosed the targets affected by Gα<SUB>12</SUB> gene knockout. An integrative network of miRNAs and mRNA changes enabled us to predict ZEB1 as a key molecule governed by Gα<SUB>12</SUB>. Decreases of miR-200a/b, -192 and -215 by Gα<SUB>12</SUB> caused ZEB1 induction. The ability of Gα<SUB>12</SUB> to decrease p53 levels, as a result of activating protein-1 (AP-1)/c-Jun-mediated mouse double minute 2 homolog induction, contributed to transcriptional deregulation of the miRNAs. Gα<SUB>12</SUB>QL induced ZEB1 and other epithelial–mesenchymal transition markers with fibroblastoid phenotype change. Consistently, transfection with miR-200b, -192 or -215 mimic prevented the ability of Gα<SUB>12</SUB>QL to increase tumor cell migration/invasion. In xenograft studies, sustained knockdown of Gα<SUB>12</SUB> decreased the overall growth rate and average volume of tumors derived from SK-Hep1 cell (mesenchymal-typed). In HCC patients, miR-192, -215 and/or -200a were deregulated with microvascular invasion or growth advantage. In the HCC samples with higher Gα<SUB>12</SUB> level, a correlation existed in the comparison of relative changes of Gα<SUB>12</SUB> and ZEB1. In conclusion, Gα<SUB>12</SUB> overexpressed in HCC causes ZEB1 induction by deregulating p53-responsive miRNAs, which may facilitate epithelial–mesenchymal transition and growth of liver tumor. These findings highlight the significance of Gα<SUB>12</SUB> upregulation in liver tumor progression, implicating Gα<SUB>12</SUB> as an attractive therapeutic target.

Gut commensal Bacteroides acidifaciens prevents obesity and improves insulin sensitivity in mice

Yang, J-Y,Lee, Y-S,Kim, Y,Lee, S-H,Ryu, S,Fukuda, S,Hase, K,Yang, C-S,Lim, H S,Kim, M-S,Kim, H-M,Ahn, S-H,Kwon, B-E,Ko, H-J,Kweon, M-N Society for Mucosal Immunology 2017 Mucosal immunology Vol.10 No.1

<P>In humans, the composition of gut commensal bacteria is closely correlated with obesity. The bacteria modulate metabolites and influence host immunity. In this study, we attempted to determine whether there is a direct correlation between specific commensal bacteria and host metabolism. As mice aged, we found significantly reduced body weight and fat mass in Atg7(Delta CD11c) mice when compared with Atg7(f/f) mice. When mice shared commensal bacteria by cohousing or feces transfer experiments, body weight and fat mass were similar in both mouse groups. By pyrosequencing analysis, Bacteroides acidifaciens (BA) was significantly increased in feces of Atg7(Delta CD11c) mice compared with those of control Atg7(f/f) mice. Wild-type C57BL/6 (B6) mice fed with BA were significantly more likely to gain less weight and fat mass than mice fed with PBS. Of note, the expression level of peroxisome proliferator-activated receptor alpha (PPARa) was consistently increased in the adipose tissues of Atg7(Delta CD11c) mice, B6 mice transferred with fecal microbiota of Atg7(Delta CD11c) mice, and BA-fed B6 mice. Furthermore, B6 mice fed with BA showed elevated insulin levels in serum, accompanied by increased serum glucagon-like peptide-1 and decreased intestinal dipeptidyl peptidase-4. These finding suggest that BA may have potential for treatment of metabolic diseases such as diabetes and obesity.</P>

NCKX3 was compensated by calcium transporting genes and bone resorption in a NCKX3 KO mouse model

Yang, H.,Ahn, C.,Shin, E.K.,Lee, J.S.,An, B.S.,Jeung, E.B. North-Holland 2017 Molecular and cellular endocrinology Vol.454 No.-

<P>Gene knockout is the most powerful tool for determination of gene function or permanent modification of the phenotypic characteristics of an animal. Existing methods for gene disruption are limited by their efficiency, time required for completion and potential for confounding off-target effects. In this study, a rapid single-step approach to knockout of a targeted gene in mice using zinc-finger nucleases (ZFNs) was demonstrated for generation of mutant (knockout; KO) alleles. Specifically, ZFNs to target the sodium/calcium/potassium exchanger3 (NCKX3) gene in C57b1/6j were designed using the concept of this approach. NCKX3 KO mice were generated and the phenotypic characterization and molecular regulation of active calcium transporting genes was assessed when mice were fed different calcium diets during growth. General phenotypes such as body weight and plasma ion level showed no distinct abnormalities. Thus, the potassium/sodium/calcium exchanger of NCKX3 KO mice proceeded normally in this study. As a result, the compensatory molecular regulation of this mechanism was elucidated. Renal TRPV5 mRNA of NCKX3 KO mice increased in both male and female mice. Expression of TRPV6 mRNA was only down regulated in the duodenum of male KO mice. Renal- and duodenal expression of PTHR and VDR were not changed; however, GR mRNA expression was increased in the kidney of NCKX3 KO mice. Depletion of the NCKX3 gene in a KO mouse model showed loss of bone mineral contents and increased plasma parathyroid hormone, suggesting that NCKX3 may play a role in regulating calcium homeostasis. (C) 2017 Elsevier B.V. All rights reserved.</P>

Plant regeneration via adventitious bud formation from cotyledon explants of Panax ginseng C.A. Meyer

Yang, D.C,Choi, K.T,Yoon, E.S,Choi, Y.E 公州大學校 基礎科學硏究所 1998 自然科學硏究 Vol.7 No.-

Cotyledon explants of ginseng (Panax ginseng C. A. Meyer) produced somatic embryos directly on medium without growth regulators, with 89% of the explants forming somatic embryos. Cytokinin treatment greatly suppressed somatic embryo formation but stimulated the direct formation of adventitious bu.ls. BAP treatment was more effective than the kinetin treatment for adventitious bud formation. Auxin (0.05 mg/1 IBA) in combination with cytokinin enhanced adventitious bud formation, with the highest frequency, 40%, at 0.05 mg/l IBA and 5 mg/l BAP. Adventitious buds were mainly formed near the distal portion of the cotyledons, while somatic embryos were formed near the proximal excised margins. Shoots were developed from adventitious buds after transfer to MS medium with 10 mg/l GA3. Root formation from the shoots was obtained after the shoots were transferred to half-strength MS medium with auxin (IAA). When the plants derived from adventitious buds were transferred to greenhouse soil, 36% were successfully acclimatized.

Targeting Non-proteolytic Protein Ubiquitination for the Treatment of Diffuse Large B Cell Lymphoma

Yang, Y.,Kelly, P.,Shaffer, A.L.,Schmitz, R.,Yoo, H.M.,Liu, X.,Huang, D.W.,Webster, D.,Young, R.M.,Nakagawa, M.,Ceribelli, M.,Wright, G.W.,Yang, Y.,Zhao, H.,Yu, X.,Xu, W.,Chan, W.C.,Jaffe, E.S.,Gascoy Cell Press 2016 CANCER CELL Vol.29 No.4

Chronic active B cell receptor (BCR) signaling, a hallmark of the activated B cell-like (ABC) subtype of diffuse large B cell lymphoma (DLBCL), engages the CARD11-MALT1-BCL10 (CBM) adapter complex to activate IκB kinase (IKK) and the classical NF-κB pathway. Here we show that the CBM complex includes the E3 ubiquitin ligases cIAP1 and cIAP2, which are essential mediators of BCR-dependent NF-κB activity in ABC DLBCL. cIAP½ attach K63-linked polyubiquitin chains on themselves and on BCL10, resulting in the recruitment of IKK and the linear ubiquitin chain ligase LUBAC, which is essential for IKK activation. SMAC mimetics target cIAP½ for destruction, and consequently suppress NF-κB and selectively kill BCR-dependent ABC DLBCL lines, supporting their clinical evaluation in patients with ABC DLBCL.

E/D-GaAs MESFET의 문턱전압 변동에 대한 T-Flipflop의 수율 시뮬레이션

김지환(J.H. Kim),윤경식(K.S. Yoon),양전욱(J.W. Yang),오응기(E.G. Oh),박철순(C.S. Park) 대한전자공학회 1996 대한전자공학회 학술대회 Vol.1996 No.11

Fabrication of spherical biochar by a two-step thermal process from waste potato peel

Yang, Xiao,Kwon, Eilhann E.,Dou, Xiaomin,Zhang, Ming,Kim, Ki-Hyun,Tsang, Daniel C.W.,Ok, Yong Sik Elsevier 2018 Science of the Total Environment Vol.626 No.-

<P><B>Abstract</B></P> <P>The aim of this study was to develop a new approach for the preparation of spherical biochar (SBC) by employing a two-step thermal technology to potato peel waste (PPW). Potato starch (PS), as a carbon-rich material with microscale spherical shape, was separated from PPW as a precursor to synthesizing SBC. The synthesis process comprised (1) pre-oxidization (preheating under air) of PS at 220 °C and (2) subsequent pyrolysis of the pretreated sample at 700 °C. Results showed that the produced SBC successfully retained the original PS morphology and that pre-oxidization was the key for its shape maintenance, as it reduced surface tension and enhanced structural stability. The SBC possessed excellent chemical inertness (high aromaticity) and uniform particle size (10–30 μm). Zero-cost waste material with a facile and easy-to-control process allows the method to be readily scalable for industrialization, while offering a new perspective on the full use of PPW.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Potato starch (PS) derived from potato peel waste (PPW) has highly regular spherical shape. </LI> <LI> PS morphology was retained with pre-oxidation process after pyrolysis. </LI> <LI> PS's spherical shape affords great homogeneity to the produced biochar. </LI> <LI> Mechanism of shape maintenance was studied via various spectral characterizations. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>