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Targeting tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) signaling is a promising approach in cancer treatment. Although ERK and/or NF-κB signaling is involved in the expression of TRAIL receptors (TRAIL-R), the exact underlying mechanisms remain unknown. In this study, we evaluated the role of ERK2 and NF-κB in the cytotoxicity of TRAIL during cisplatin treatment. Cisplatin treatment of neuroepithelioma cells (SK-N-MC) significantly induced ERK2 activation and increased TRAIL cytotoxicity via the upregulation of death receptor 5 (DR5) expression. In partial ERK2 knockdown cell lines that maintained only basal levels of ERK2 activity, cisplatin treatment did not increase ERK2 activity or DR5 expression. These findings indicate that induced (rather than basal) ERK2 activity enhances TRAIL susceptibility via DR5 expression. In complete ERK2 knockdown cell lines with no basal ERK2 activity, DR4, DR5, and DcRs expression levels were increased, and additional treatment with cisplatin did not further increase TRAIL-R expression. Chemical inhibition of ERK2 also enhanced TRAIL cytotoxicity by upregulating DR4 and DR5 expression. These findings indicate that basal ERK2 activity suppresses TRAIL-R expression. Both basal and inducible ERK2 activities regulate TRAIL-R expression via the NF-κB signaling pathway. Overall, our findings suggest that the ERK2/NF-κB signaling pathway has a dual role in TRAIL susceptibility by differentially regulating TRAIL-R expression in the same cellular system.

Chronic airway exposure to harmful substances, such as deleterious gases, cigarette smoke (CS), and particulate matter, triggers chronic obstructive pulmonary disease (COPD), characterized by impaired lung function and unbridled immune responses. Emerging epigenomic and genomic evidence suggests that excessive recruitment of alveolar macrophages and neutrophils contributes to COPD pathogenesis by producing various inflammatory mediators, such as reactive oxygen species (ROS), neutrophil elastase, interleukin (IL) 6, and IL8. Recent studies showed that Epilobium species attenuated ROS, myeloperoxidase, and inflammatory cytokine production in murine and human innate immune cells. Although the Epilobium genus exerts anti-inflammatory, antioxidant, and antimicrobial effects, the question of whether the Epilobium species regulate lung inflammation and innate immune response in COPD has not been investigated. In this study, Epilobium pyrricholophum extract (EPE) suppressed inflammatory cell recruitment and clinical symptoms in porcine pancreatic elastase and CS extract-induced COPD mice. In addition, EPE attenuated inflammatory gene expression by suppressing MAPKs and NFκB activity. Furthermore, UPLC-Q-TOF MS analyses revealed the anti-inflammatory effects of the identified phytochemical constituents of EPE. Collectively, our studies revealed that EPE represses the innate immune response and inflammatory gene expression in COPD pathogenesis in mice. These findings provide insights into new therapeutic approaches for treating COPD.

Allergic rhinitis (AR) is an allergic nasal disease characterized by nasal obstruction, rhinorrhea, sneezing, and itching. Type 1 helper T cells (Th1)/type 2 helper T cells (Th2) imbalance has been identified as an important immunological mechanism of AR. In addition, up-regulation of type 17 helper T cells (Th17) also increase the risk of developing AR. Gallic acid (3, 4, 5-trihydroxybenzoic acid, GA), a polyphenol natural product, is obtained from various herbs, red wine, and green tea. It is known to have diverse biological effects such as anti-oxidation, anti-inflammation, anti-microbial and anti-cancer. In the present study, the effect of GA on airway inflammation and expression of Th1, Th2 and Th17 cytokines in an ovalbumin (OVA)-induced AR mouse model were investigated. GA alleviated the nasal allergic symptoms, reduced the thickness of nasal mucosa, attenuated goblet cell hyperplasia and eosinophil cell infiltration in the nasal mucosa, decreased the levels of interleukin (IL)-4, IL-5, IL-13 and IL-17 in nasal lavage fluid (NALF), and diminished the levels of OVA-specific IgE, OVA-specific IgG1 and OVA-specific IgG2a in serum. However, GA increased the expression of interferon-gamma and IL-12 in NALF. Taken together, it suggests that GA may be used as a therapeutic agent for AR.

Ethnopharmacological relevance Dryopteris crassirhizoma (DC) is used as a traditional herbal remedy to treat various diseases, the tapeworm infection, common cold, and cancer in Korea, Japan, and China. DC also has the antioxidant anti-inflammatory and antibacterial activities. However, the anti-allergic inflammatory effect of DC and some of its mechanisms in allergic rhinitis model are unknown well. Aim of this study The purpose of this study is to investigate the anti-allergic inflammatory effect of DC on the allergic rhinitis model, mast cell activation and histamine release. Materials and methods Allergic rhinitis was induced in BALB/c mice by sensitization and challenge with ovalbumin (OVA). Different concentration of DC and dexamethasone was administrated by oral gavage on 1 h before the OVA challenge. Mice of the control group were treated with saline only. Then mice were evaluated for the presence of nasal mucosa inflammation, the production of allergen-specific cytokine response and the histology of nasal mucosa. Results DC significantly ameliorated the nasal symptoms and the inflammation of nasal mucosa. DC also reduced the infiltration of eosinophils and mast cells in these tissues and the release of histamine in blood. Meanwhile, DC evidently inhibited the overproduction of Th2 cytokines and increased the Th1 and Treg cytokines in nasal lavage fluid by OVA. DC also reduced the levels of OVA-specific IgE, IgG1 and IgG2a in blood. Conclusions This study suggests that DC has a significant anti-allergic inflammatory effect in the nasal cavity. DC may have the therapeutic effect of allergic rhinitis.

A series of chalcone derivatives were synthesized and evaluated for their μ-calpain and cathepsin B inhibitory activities. Among the tested chalcone derivatives, two compounds, 7 and 11, showed potent inhibitory activities against μ-calpain and cathepsin B and were selected for further evaluation. Compounds 7 and 11 showed enzyme inhibitory activities at the cellular level and displayed neuroprotective effects against oxidative stress-induced apoptosis in SH-SY5Y cells, a human neuroblastoma cell line. Moreover, compounds 7 and 11 reduced p25 formation, tau phosphorylation and insoluble Aβ peptide formation. Enzyme kinetic experiments and docking studies revealed that compounds 7 and 11 competitively inhibited both μ-calpain and cathepsin B enzymes.

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), either alone or in combination with other anti-cancer agents, is a promising new strategy for the treatment of cancer. However, aberrant PI3K/Akt/mTOR survival signaling may confer TRAIL resistance by altering the balance between pro- and anti-apoptotic proteins. In the present study, we showed that the Akt/mTOR inhibitor RAD001 (everolimus) induced cell death in a dose-dependent manner and enhanced TRAIL-induced apoptosis in human leukemic Jurkat T cells, which show PI3K/Akt/mTOR pathway activation and basal expression levels of death receptor (DR) 5 (TRAIL-R2). Investigation of the effect of RAD001 treatment on the expression of TRAIL receptors (TRAIL-Rs) in Jurkat T cells showed that RAD001 significantly upregulated DR5 by up to 51.22%, but not other TRAIL-Rs such as DR4 (TRAIL-R1), decoy receptor (DcR) 1 (TRAIL-R3), and DcR2 (TRAIL-R4). Pretreatment with DR5:Fc chimera abrogated the RAD001-induced increase of TRAIL cytotoxicity, indicating that the upregulation of DR5 by RAD001 plays a role in enhancing the susceptibility of Jurkat T cells to TRAIL. Our results indicate that combination treatment with RAD001 and TRAIL may be a novel therapeutic strategy in leukemia.

Recombinant DNA pLuc2 which contains a firefly luciferase cDNA from the commercial pGL3-Basic plasmid vector was constructed for bioluminescent Escherichia coli and lactobacilli. The bioluminescent signal strengths of E. coli DH5α and L. casei CJNU 0588 transformed with pLuc2 were 2.78×109 and 1.19×106 p/s, respectively. Approximately 1010 cells of bioluminescent L. casei CJNU 0588 (pLuc2) were given orally to 6-week-old female Balb/c mice. The bioluminescent signal was detected from all three mice at 1 h and one mouse at 3 h but was not detected thereafter. The signal was also confirmed from an opened abdomen and the extracted GI tract of a mouse at 1 h. From the results, we have known that L. casei CJNU 0588 pass through the murine stomach and reach the colon and cecum 1 h after oral administration.

Deduplication, which is a technique of eliminating redundant da ta by storing only a single copy of each data, provides clients and a cloud server with efficiency for managing stored data. Si nce the data is saved in untrusted public cloud server, however , both invasion of data privacy and data loss can be occurred. Ov er recent years, although many studies have been proposed secure deduplication schemes, there still remains both the secu rity problems causing serious damages and inefficiency.In this paper, we propose secure and efficient client-side dedu plication with Key-server based on Bellare et. al’s scheme and challenge-response method. Furthermore, we point out potential risks of client-side deduplication and show that our scheme is secure against various attacks and provides high efficiency for uploading big size of data. Keywords: Cloud storage, Client-side deduplication, Data privacy, Poison attack I.서 론 1) 1.1개요빅 데이터(Big data) 시대를 맞아 클라이언트들 접수일(2014년 10월 15일), 수정일(2014년 12월 22일),게재확정일(2015년 1월 28일)* 이 논문은 2012년도 정부(교육과학기술부)의 재원으로 한국연구재단의 지원을 받아 수행된 연구임(No.2012-00

Although there has been a steady increase in the prevalence of food allergies worldwide in recent decades, no effective therapeutic strategies have been developed. Modulation of the gut microbiota composition and/or function through probiotics has been highlighted as a promising target for protection against food allergies. In this study, we aimed to investigate the allergy-reducing effects of a probiotic mixture (P5: Lactococcus lactis KF140, Pediococcus pentosaceus KF159, Lactobacillus pentosus KF340, Lactobacillus paracasei 698, and Bacillus amyloliquefaciens 26N) in mice with ovalbumin (OVA)-induced food allergy. Administration of P5 significantly suppressed the oral OVA challenge-induced anaphylactic response and rectal temperature decline, and reduced diarrhea symptoms. Moreover, P5 also significantly inhibited the secretion of IgE, Th2 cytokines (interleukin (IL)-4, IL-5, IL-10, and IL-13), and Th17 cytokines (IL-17), which were increased in mice with OVA-induced food allergy, and induced generation of CD4+Foxp3+ regulatory T cells. These results revealed that P5 may have applications as a preventive agent against food allergy.

Myocardial ischemia (MI) induces many changes in the body, including pH decrease and electrolyte imbalance. No obvious symptoms of MI appear until irreversible cellular injuries occur. Since early treatment is critical for recovery from ischemia, the development of reliable diagnostic tool is demanded to detect the early ischemic status. Ischemia modified albumin (IMA), formed by cleavage of the last two amino acids of the human serum albumin (HSA) N-terminus, has been considered so far as the most trustworthy and accurate marker for the investigation of ischemia. IMA levels are elevated in plasma within a few minutes of ischemic onset, and may last for up to 6 h. In the present study, we developed a novel assay for the examination of IMA levels to ameliorate the known albumin cobalt binding (ACB) test established previously. We observed a stronger copper ion bound to the HSA N-terminal peptide than cobalt ion by HPLC and ESI-TOF mass spectrometric analyses. The copper ion was employed with lucifer yellow (LY), a copper-specific reagent to develop a new albumin copper binding (ACuB) assay. The parameters capable of affecting the assay results were optimized, and the finally-optimized ACuB assay was validated. The result of the IMA level measurement in normal versus stroke rat serum suggests that the ACuB assay is likely to be a reliable and sensitive method for the detection of ischemic states.

Pediocin PA-1, mainly produced from Pediococcus spp., is a well-known class IIa bacteriocin showing strong anti-listerial activity. Pediococcus acidilactici K10 producing the bacteriocin was transformed with pLR5cat(S)_PSAB in which the pediocin PA-1 structural gene (pedA) was translationally fused with a deduced signal sequence of the bifidobacterial α-amylase gene for secretion. The P. acidilactici K10 transformant presented increased bacteriocin activity when compared with that of native P. acidilactici K10. The P. acidilactici K10 mutant (K10M), in which the plasmid for pediocin PA-1 production is cured, also showed bacteriocin activity when transformed with pLR5cat(S)_PSAB, indicating that the pediocin gene was expressed in hosts. An increase in pedA gene transcription level and the detection of both transcripts from the pediocin operon and PSAB were also confirmed. The P. acidilactici K10 [pLR5cat(S)_PSAB] transformant was more inhibitory against Listeria monocytogenes than that of the P. acidilactici K10 wild type in co-cultures.

The key neuropathological features of Alzheimer's disease are abnormal deposition of Aβ plaques and insoluble Aβ peptides in extracellular brain and intracellular neurofibril tangles induced by abnormal tau hyperphosphorylation. μ-Calpain is one of the factors that bridge these Aβ- and hyperphosphorylated tau-mediated pathological pathways. Undecylenic acid (UDA), a naturally occurring unsaturated fatty acid, was discovered as a μ-calpain inhibitor by screening a chemical library using a substrate specific μ-calpain assay method. UDA inhibited Aβ oligomerization and Aβ fibrillation and reversed Aβ-induced neuronal cell death. In addition, UDA scavenged ROS and reversed the levels of proapoptotic proteins induced by ROS in SH-SY5Y cells. UDA inhibited μ-calpain activity with better potency than the known peptide-like μ-calpain inhibitor, MDL28170, in SH-SY5Y and HEK293T cells transfected with the catalytic subunit of μ-calpain. These results suggest that UDA is a novel non-peptide-like μ-calpain inhibitor with good cell permeability and potent neuroprotective effect.

To support beneficial effects of makgeolli for human health, we investigated for the presence of 1,4-dihydroxy-2-naphthoic acid (DHNA), a bifidogenic growth stimulator (BGS), from commercial makgeolli products. Among eleven makgeolli products (A∼K), four showed positive peaks for DHNA in high performance liquid chromatography analysis. Makgeolli product A in particular contained the highest concentration of DHNA (0.44 ppm), as confirmed by liquid chromatography-mass spectrometry. Furthermore, BGS activity of the makgeolli product A was higher than those of products in which DHNA was not detected. These results indicate that makgeolli can be a good source for DHNA and that DHNA-enriched makgeolli could be developed by modifying manufacturing procedures and controlling its microbiota.

Previous studies have confirmed that fermented whey produced by Leuconostoc mesenteroides CJNU 0147 or Lactobacillus casei CJNU 0588 display bifidogenic growth stimulator (BGS) activity. The present study sought to determine if the strain itself can improve the growth of bifidobacteria in co-cultures. In reinforced clostridial medium (RCM), both strains stimulated the growth of a Bifidobacterium lactis strain during the exponential phase and also stimulated the growth during almost all growth phases in whey broth. Fermented whey containing viable Leu. mesenteroides CJNU 0147 and L. casei CJNU 0588 cells maintained viability of the B. lactis strain stored at 10℃ in MRS broth. Viable cell count of the B. lactis strain without the fermented whey was decreased to 5.6 log cfu/mL after 15 days, whereas that of the strain with the fermented whey was slightly increased to 7.1 log cfu/mL as compared with initial viable cell count of 6.9 log cfu/mL.

Various culture conditions for the production of fermented whey presenting bifidogenic growth stimulator (BGS) activity were evaluated using Leuconostoc mesenteroides CJNU 0147 and Lactobacillus casei CJNU 0588. The BGS activity of fermented whey produced with mixed culture of Leu. mesenteroides CJNU 0147 and L. Casei CJNU 0588 was higher than those of single cultures. The optimal temperature for the production of the fermented whey was 20°C. The anaerobic culture conditions via nitrogen gas supply had no influence on the BGS activity of fermented whey. The BGS activity of the heat-treated fermented whey samples was slightly decreased by 7.63, 11.66, and 15.12% at 80, 100, and 121°C, respectively for 15 min. Pilot-scale (75 L) fermented whey was produced using the 2 freeze-dried cell powders of CJNU 0147 and CJNU 0588 and spray-dried. The spray-dried fermented whey presented BGS activity, indicating it can be used as a functional food material.

In order to extend the scaffold of non-peptidic calpain inhibitor, we have designed and synthesized 14 chalcone derivatives categorized into two groups based on their structures. Compounds 7 () and 8 () in group A were most selective -calpain inhibitor over cathepsins B and L. On the other hand, compound 14 possessing furan ring exhibited inhibitory activities for -calpain () as well as cathepsin B (). The results discovered implicated that chalcone analogues possessing proper size and functional groups can be a potential lead core for selective non-peptidic -calpain inhibitor. Furthermore, dual inhibitors for -calpain and cathepsin B can also be developed from chalcones by elaborate structure manipulation.

The recombinant DNA pLR5cat_PSAB in which pediocin PA-1 structural and immunity genes (pedAB) fused with the promoter and deduced signal sequence of an alpha-amylase gene from a bifidobacterial strain were inserted in pLR5cat, an Escherichia coli-lactobacilli shuttle vector was transferred to Lactobacillus reuteri KCTC 3679 and the transformant presented bacteriocin activity. The recombinant L. reuteri KCTC 3679 transformed with the shortened pLR5cat(S)_PSAB, where non-essential region for the lactobacilli replicon was removed, also showed bacteriocin activity. The molecular mass of the secreted pediocin PA-1 from the recombinant bacteria was the same as that of native pediocin PA-1 (~4.6 kDa) from Pediococcus acidilactici K10 on a sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gel. In co-cultures with Listeria monocytogenes, the recombinant L. reuteri KCTC 3679 effectively reduced the viable cell count of the pathogenic bacterium by a 3 log scale compared with a control where L. monocytogenes was incubated alone.

Several strains of Penicillium section Citrina were isolated during a survey of fungi from marine environments along the southern coast of Korea. Based on multigene phylogenetic analyses (β-tubulin and calmodulin) and morphological characteristics, the 11 strains were identified as P. citrinum, P. hetheringtonii, P. paxilli, P. sumatrense, P. terrigenum, and P. westlingii. To understand the ecological role of these species, we tested all strains for extracellular enzyme activity; six strains representing four species showed β-glucosidase activity. Four of the identified species — P. hetheringtonii, P. paxilli, P. terrigenum, and P. westlingii — are new records for Korea. For these new species records, we describe morphological characteristics of the strains and compare results to published data of type strains.

Human serum albumin (HSA) is the most abundant protein in the human body. HSA injections prepared by fractionating human blood have mainly covered the demand for albumin to treat hypoalbuminemia, the state of low concentration of albumin in blood. HSA in solution may exist in various forms such as monomers, oligomers, polymers, or as mixtures, and its conformational change and/or aggregation may occur easily. Considering these characteristics, there is a great chance of modification and polymer formation during the preparation processes of albumin products, especially injections. The albumin cobalt binding (ACB) test reported by Bar-Or et al. was originally designed to detect ischemia modified albumin (IMA), which contains the modified HSA N-terminal sequence by cleavage of the last two amino acids. In this study, we developed a cobalt albumin binding (CAB) assay to correct the flaws of the ACB test with improving the sensitivity and precision. The newly developed CAB assay easily detects albumin configuration alterations and may be able to be used in developing a quality control method for albumin and its pharmaceutical formulations including albumin injections.

μ-Calpain is a calcium-dependent cysteine protease, which is activated by μM concentration of calcium in vitro. Disrupted intracellular calcium homeostasis leads to hyper-activation of μ-calpain. Hyper-activated μ-calpain enhances the accumulation of β-amyloid peptide by increasing the expression level of β-secretase (BACE1) and induces hyper-phosphorylation of tau along with the formation of neurofibrillary tangle by mediating p35 cleavage into p25, both of which are the major mechanisms of neurodegeneration in Alzheimer's disease (AD). Hence, inhibition of μ-calpain activity is very important in the treatment and prevention of AD. In this study, conjugated linoleic acid (CLA), an eighteen-carbon unsaturated fatty acid, was discovered as a μ-calpain-specific inhibitor. CLA showed neuroprotective effects against neurotoxins such as H2O2 and Aβ1-42 in SH-SY5Y cells, and inhibited Aβ oligomerization/fibrillation and Aβ-induced Zona Occludens-1 degradation. In addition, CLA decreased the levels of proapoptotic proteins, p35 conversion to p25 and tau phosphorylation. These findings implicate CLA as a new core structure for selective μ-calpain inhibitors with neuroprotective effects. CLA should be further evaluated for its potential use as an AD therapeutic agent.

A new procedure for the preparation of emodin carbaldehyde and citreorosein was described, in which, ω,ω'-dibromomethylemodin triacetate was prepared as a key intermediate by NBSmediated bromination of 1,3,8-triacetylemodin. Reduction of emodin and citreorosein with SnCl(2) in a 1:1 mixture of HOAc and HCl afforded the corresponding anthrones in 90% and 92% yield, respectively, while the corresponding 10-desoxyemodin carbaldehyde was prepared by MnO(2) oxidation of 10-desoxycitreorosein. 10-Desoxycitreorosein and emodin carbaldehyde showed feasible μ-calpain inhibitory activities with IC(50) values of 20.15 and 25.77 M, respectively.

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a new promising agent for clinical use since it kills a wide range of tumor cells but not normal cells. It is regulated by 2 death receptors (DR), DR4 (TRAIL-R1) and DR5 (TRAIL-R2), and 2 decoy receptors (DcR), DcR1 (TRAIL-R3) and DcR2 (TRAIL-R4). However, the determining factors of the sensitivity to TRAIL cytotoxicity are not clearly understood. This study aims to elucidate the role of extracellular signal-regulated kinase (ERK) on enhancement of TRAIL cytotoxicity in cultured neuroepithelioma cell line SK-N-MC. When 1 μg/ml cisplatin-pretreated cells were exposed to 100 ng/ml TRAIL for 48 h, apoptotic cell death increased, compared with the cells treated with cisplatin or TRAIL alone (66% versus 27% or 9% apoptotic cell death). In addition, pretreatment with cisplatin in SK-N-MC cells significantly increased ERK1/2 activity and increased expression levels of DR5, but not DR4, DcR1 and DcR2, which correlated with increase of their susceptibility to TRAIL cytotoxicity. Introduction of ERK2 siRNA or ERK2 shRNA in SK-N-MC cells significantly decreased cisplatin-induced DR5 expression, suggesting that cisplatin-induced ERK2 activity is involved in DR5 expression. On the other hand, treatment with 20 μM U0126, an inhibitor of MEK, completely reduced basal ERK1/2 activity and induced cell death in SK-N-MC cells. In addition, pretreatment with U0126 increased TRAIL-mediated apoptosis via up-regulation of DR5 expression in response to the decrease in basal ERK2 activity. Because pretreatment with U0126 completely inhibited not only cisplatin-induced ERK2 activation but also basal ERK2 activity, the following treatment with TRAIL significantly increased cell death, indicating that constitutively activated ERK2 blocks DR5 expression in SK-N-MC cells. Taken together, our results implicate that basal ERK2 activity is involved in TRAIL resistance but induced ERK2 plays a role in TRAIL sensitivity, and suggest that the enhanced sensitivity of SK-N-MC cells to TRAIL is associated with an increase in DR5 expression. Citation Format: Hye Lim Jung, Myoung Woo Lee, Ji Eun Eom, Dea Seong Kim, Hyo-Jung Park, Dong-Kyung Yu, Meong Hi Son, Soo Hyun Lee, Keon Hee Yoo, Ki Woong Sung, Hong Hoe Koo. Dual roles of ERK2 to TRAIL sensitivity in SK-N-MC neuroepithelioma cells. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2937. doi:10.1158/1538-7445.AM2013-2937