Acanthopanax koreanum Fruit Waste Inhibits Lipopolysaccharide-Induced Production of Nitric Oxide and Prostaglandin E2 in RAW 264.7 Macrophages

The Acanthopanax koreanum fruit is a popular fruit in Jeju Island, but the byproducts of the alcoholic beverage prepared using this fruit are major agricultural wastes. The fermentability of this waste causes many economic and environmental problems. Therefore, we investigated the suitability of using A. koreanum fruit waste (AFW) as a source of antiinflammatory agents. AFWs were extracted with 80% EtOH. The ethanolic extract was then successively partitioned with hexane, CH2Cl2, EtOAc, BuOH, and water. The results indicate that the CH2Cl2 fraction (100 μg/mL) of AFW inhibited the LPS-induced nitric oxide (NO) and prostaglandin E2 (PGE2) production in RAW 264.7 cells by 79.6% and 39.7%, respectively. These inhibitory effects of the CH2Cl2 fraction of AFWs were accompanied by decreases in the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) proteins and iNOS and COX-2 mRNA in a dose-dependent pattern. The CH2Cl2 fraction of AFWs also prevented degradation of IκB-α in a dose-dependent manner. Ursolic acid was identified as major compound present in AFW, and CH2Cl2 extracts by high performance liquid chromatography (HPLC). Furthermore using pure ursolic acid as standard and by HPLC, AFW and CH2Cl2 extracts was found to contain 1.58 mg/g and 1.75 mg/g, respectively. Moreover, we tested the potential application of AFW extracts as a cosmetic material by performing human skin primary irritation tests. In these tests, AFW extracts did not induce any adverse reactions. Based on these results, we suggest that AFW extracts be considered possible anti-inflammatory candidates for topical application

Hypochoeris radicata attenuates LPS-induced inflammation by suppressing P38, ERK, and JNK phosphorylation in Raw 264.7 macrophages

Hypochoeris radicata, an invasive plant species, is a large and growing threat to ecosystem integrity on Jeju Island, a UNESCO World Heritage site. Therefore, research into the utilization of H. radicata is important and urgently required in order to solve this invasive plant problem in Jeju Island. The broader aim of our research is to elucidate the biological activities of H. radicata, which would facilitate the conversion of this invasive species into high value added products. The present study was undertaken to identify the pharmacological effects of H. radicata flower on the production of inflammatory mediators in macrophages. The results indicate that the ethyl acetate fraction of H. radicata extract (HRF-EA) inhibited the production of pro-inflammatory molecules such as NO, iNOS, PGE2, and COX-2, and cytokines such as TNF-α, IL-1β, and IL-6 in LPS-stimulated RAW 264.7 cells. Furthermore, the phosphorylation of MAPKs such as p38, ERK, and JNK was suppressed by HRF-EA in a concentration-dependent manner. In addition, through HPLC and UPLC fingerprinting, luteolins were also identified and quantified as extract constituents. On the basis of these results, we suggest that H. radicata may be considered possible anti-inflammatory candidates for pharmaceutical and/or cosmetic applications

An efficient strategy for cell-based antibody library selection using an integrated vector system

BACKGROUND: Cell panning of phage-displayed antibody library is a powerful tool for the development of therapeutic and imaging agents since disease-related cell surface proteins in native complex conformation can be directly targeted. Here, we employed a strategy taking advantage of an integrated vector system which allows rapid conversion of scFv-displaying phage into scFv-Fc format for efficient cell-based scFv library selection on a tetraspanin protein, CD9. RESULTS: A mouse scFv library constructed by using a phagemid vector, pDR-D1 was subjected to cell panning against stable CD9 transfectant, and the scFv repertoire from the enriched phage pool was directly transferred to a mammalian cassette vector, pDR-OriP-Fc1. The resulting constructs enabled transient expression of enough amounts of scFv-Fcs in HEK293E cells, and flow cytometric screening of binders for CD9 transfectant could be performed simply by using the culture supernatants. All three clones selected from the screening showed correct CD9-specificity. They could immunoprecipitate CD9 molecules out of the transfectant cell lysate and correctly stain endogenous CD9 expression on cancer cell membrane. Furthermore, competition assay with a known anti-CD9 monoclonal antibody (mAb) suggested that the binding epitopes of some of them overlap with that of the mAb which resides within the large extracellular loop of CD9. CONCLUSIONS: This study demonstrates that scFv-Fc from mammalian transient expression can be chosen as a reliable format for rapid screening and validation in cell-based scFv library selection, and the strategy described here will be applicable to efficient discovery of antibodies to diverse cell-surface targets

Traffic Convexity Aware Cellular Networks: A Vehicular Heavy User Perspective

Rampant mobile traffic increase in modern cellular networks is mostly caused by large-sized multimedia contents. Recent advancements in smart devices as well as radio access technologies promote the consumption of bulky content, even for people in moving vehicles, referred to as vehicular heavy users. In this article the emergence of vehicular heavy user traffic is observed by field experiments conducted in 2012 and 2015 in Seoul, Korea. The experiments reveal that such traffic is becoming dominant, captured by the 8.62 times increase in vehicular heavy user traffic while the total traffic increased 3.04 times. To resolve this so-called vehicular heavy user problem (VHP), we propose a cell association algorithm that exploits user demand diversity for different velocities. This user traffic pattern is discovered first by our field trials, which is convex-shaped over velocity, i.e. walking user traffic is less than stationary or vehicular user traffic. As the VHP becomes severe, numerical evaluation verifies the proposed user convexity aware association outperforms a well-known load balancing association in practice, cell range expansion (CRE). In addition to the cell association, several complementary techniques are suggested in line with the technical trend toward 5G.Comment: 15 pages, 5 figures, 1 table, to appear in IEEE Wireless Communications Magazin

USP13 antagonizes gp78 to maintain functionality of a chaperone in ER-associated degradation

Physiological adaptation to proteotoxic stress in the endoplasmic reticulum (ER) requires retrotranslocation of misfolded proteins into the cytoplasm for ubiquitination and elimination by ER-associated degradation (ERAD). A surprising paradox emerging from recent studies is that ubiquitin ligases (E3s) and deubiquitinases (DUBs), enzymes with opposing activities, can both promote ERAD. Here we demonstrate that the ERAD E3 gp78 can ubiquitinate not only ERAD substrates, but also the machinery protein Ubl4A, a key component of the Bag6 chaperone complex. Remarkably, instead of targeting Ubl4A for degradation, polyubiquitination is associated with irreversible proteolytic processing and inactivation of Bag6. Importantly, we identify USP13 as a gp78-associated DUB that eliminates ubiquitin conjugates from Ubl4A to maintain the functionality of Bag6. Our study reveals an unexpected paradigm in which a DUB prevents undesired ubiquitination to sharpen substrate specificity for an associated ubiquitin ligase partner and to promote ER quality control

The prognostic factors of resected non-small cell lung cancer with chest wall invasion

<p>Abstract</p> <p>Background</p> <p>We retrospectively reviewed the clinical features and surgical outcomes of patients with a surgically resected NSCLC invading chest wall in order to identify prognostic factors that impact long term survival.</p> <p>Methods</p> <p>Between January 1990 and December 2009, 107 patients who underwent surgical resection for chest wall invading NSCLC were reviewed. Tumors invading only the parietal pleura were defined as superficial invasions, and those involving the soft tissue or ribs were defined as deep invasions.</p> <p>Results</p> <p>There were 91 men and 16 women; median age was 64 years (range 30 to 80 years). Overall 5 year survival rate was 26.3%. The univariate prognostic factors for survival included gender, extent of resection (pneumonectomy vs lobectomy), tumor size(> 5 cm vs ≤ 5 cm), nodal status (N0 or N1 vs N2), completeness of resection (complete vs incomplete) and completeness of adjuvant chemotherapy. At multivariate analysis, five independent prognostic factors were shown; depth of invasion (superficial vs deep), tumor size, nodal status, completeness of resection, and completeness of adjuvant chemotherapy. In patients with completely resected T3N0 NSCLC, completion of chemotherapy is the only prognostic factor for long term survival.</p> <p>Conclusions</p> <p>Completeness of resection, nodal status, depth of invasion, tumor size, and adjuvant chemotherapy were prognostic factors for long-term survival in NSCLC patients with chest wall invasion. Because of poor prognosis in cases with chest wall invasion that have N2 positive LN, that is difficult to achieve complete resection and that need pneumonectomy, definite chemoradiotherapy or neoadjuvant chemoradiotherapy should be considered first in these cases.</p