Search

Purpose Translocator protein 18-kDa (TSPO) positron emission tomography (PET) is a valuable tool to detect neuroinflammed areas in a broad spectrum of neurodegenerative diseases. However, the clinical application of second-generation TSPO ligands as biomarkers is limited because of the presence of human rs6971 polymorphism that affects their binding. Here, we describe the ability of a new TSPO ligand, [18F]BS224, to identify abnormal TSPO expression in neuroinflammation independent of the rs6971 polymorphism. Methods An in vitro competitive inhibition assay of BS224 was conducted with [3H]PK 11195 using membrane proteins isolated from 293FT cells expressing TSPO-wild type (WT) or TSPO-mutant A147T (Mut), corresponding to a high-affinity binder (HAB) and low-affinity binder (LAB), respectively. Molecular docking was performed to investigate the interaction of BS224 with the binding sites of rat TSPO-WT and TSPO-Mut. We synthesized a new 18F-labeled imidazopyridine acetamide ([18F]BS224) using boronic acid pinacol ester 6 or iodotoluene tosylate precursor 7, respectively, via aromatic 18F-fluorination. Dynamic PET scanning was performed up to 90 min after the injection of [18F]BS224 to healthy mice, and PET imaging data were obtained to estimate its absorbed doses in organs. To evaluate in vivo TSPO-specific uptake of [18F]BS224, lipopolysaccharide (LPS)-induced inflammatory and ischemic stroke rat models were used. Results BS224 exhibited a high affinity (Ki = 0.51 nM) and selectivity for TSPO. The ratio of IC50 values of BS224 for LAB to that for HAB indicated that the TSPO binding affinity of BS224 has low binding sensitivity to the rs6971 polymorphism and it was comparable to that of PK 11195, which is not sensitive to the polymorphism. Docking simulations showed that the binding mode of BS224 is not affected by the A147T mutation and consequently supported the observed in vitro selectivity of [18F]BS224 regardless of polymorphisms. With optimal radiochemical yield (39 ± 6.8%, decay-corrected) and purity (> 99%), [18F]BS224 provided a clear visible image of the inflammatory lesion with a high signal-to-background ratio in both animal models (BPND = 1.43 ± 0.17 and 1.57 ± 0.37 in the LPS-induced inflammatory and ischemic stroke rat models, respectively) without skull uptake. Conclusion Our results suggest that [18F]BS224 may be a promising TSPO ligand to gauge neuroinflammatory disease-related areas in a broad range of patients irrespective of the common rs6971 polymorphism.

The emergence and rapid spread of the potentially fatal coronavirus disease 2019, caused due to infection by severe acute respiratory syndrome coronavirus-2, has led to worldwide interest in developing functional bioactive ingredients that act as immunomodulatory agents. In this study, we aimed to characterize Carica papaya extract and explore its potential as an immuno-modulator by performing in vitro cell screening. Papaya leaf water extract (PLW) was found to significantly increase the levels of nitric oxide (NO) and prostaglandin E2 (PGE2) by upregulating inducible nitric oxide synthase and cyclo-oxygenase-2 activity, respectively. Additionally, PLW increased the production of tumor necrosis factor-α and interleukin 1β in RAW 264.7 cells. Furthermore, PLW activated the expression of c-Jun N-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK) but not that of p38 mitogen-activated protein kinase. These results indicate that PLW increased the production of NO, PGE2, and pro-inflammatory cytokines by activating the JNK and ERK pathways in macrophages, thus demonstrating immunomodulatory properties. Finally, high-performance liquid chromatography fingerprint analysis indicated the presence of rutin, narirutin, and ρ-coumaric acid in PLW (6.30, 119.76, and 47.25 ppm, respectively). Treating cells with these compounds at non-toxic concentrations had no effect on NO production. Taken together, these results suggest that PLW may have potential as an immunity-enhancing supplement. © The Korean Society for Applied Biological Chemistry 2021.

DHP107 is a newly developed lipid‐based oral formulation of paclitaxel. We evaluated the in vivo tissue pharmacokinetics (PKs) of DHP107 in mice and patients using positron emission tomography (PET). Radioisotope‐labeled [3H]DHP107 and [18F]DHP107 for oral administration were formulated in the same manner as the manufacturing process of DHP107. In vivo tissue PK were assessed in healthy ICR mice and breast cancer xenografted SCID mice. Two patients with metastatic breast cancer were clinically evaluated for absorption at the target lesion after internal absorbed dose estimation. Whole‐body PET/computed tomography data were acquired in healthy and xenografted mice and in patients up to 10–24 h after administration. Tissue [18F]DHP107 signals were plotted against time and PK parameters were determined. The amounts of radioactivity in various organs and excreta were determined using a beta‐counter and are expressed as the percentage of injected dose (ID). Oral [18F]DHP107 was well‐absorbed and reached the target lesion in mice and patients with breast cancer. Significant amounts of radioactivity were found in the stomach, intestine, and liver after oral administration of [3H]‐ and [18F]DHP107 in healthy mice. The [18F]DHP107 reached a peak distribution of 0.7–0.8%ID in the tumor at 5.6–7.3 h in the xenograft model. The [18F]DHP107 distribution in patients with metastatic breast cancer was the highest at 3–4 h postadministration. Systemic exposures after administration of a DHP107 therapeutic dose were comparable with those in previous studies. PET using radioisotope‐labeled drug candidates is useful for drug development and can provide valuable information that can complement plasma PK data, particularly in early phase clinical trials.

The effect of medium composition on enzyme and β-glucan production by Aspergillus oryzae KCCM 12698 was investigated. Brown rice, rice bran, nitrogen, and ascorbic acid are key components of the synthetic medium used in liquid-state fermentation. To determine the optimal concentrations of these components for enzyme and β-glucan production, we conducted one factor at a time experiments, which showed that the optimal concentrations were 30 g/l brown rice, 30 g/l rice bran, 10 g/l soytone, and 3 g/l ascorbic acid. Pretreatment of brown rice for 60 min prior to inoculation enhanced fungal biomass, while increasing the production of enzymes and β-glucan using solidstate fermentation. Maximum fungal biomass of 0.76 mg/g, amylase (26,551.03 U/g), protease (1,340.50 U/g), and β-glucan at 9.34% (w/w) were obtained during fermentation. Therefore, solidstate fermentation of brown rice is a process that could enhance yield and overall production of enzymes and β-glucan for use in various applications.

Cudraxanthone D (CD) is a natural xanthone compound derived from the root barks of Cudrania tricuspidata . However, the biological functions of CD in human metabolism have been rarely reported until now. Autophagy is the self-degradation process related to cancer cell metastasis. Here, we elucidated the effects of CD on human oral squamous cell carcinoma (OSCC) cells’ metastatic ability. We confirmed that CD effectively decreased the proliferation and viability of SCC25 human OSCC cells in time- and dose-dependent manners. Also, the metastasis phenotype of the SCC25 cell (migration, invasion, and epithelial–mesenchymal transition [EMT]) was inhibited by CD. To further investigate the mechanism by which CD inhibited the metastatic capacity, we detected the relationship between EMT and autophagy in the SCC25 cells. The results revealed that CD inhibited the metastasis of the SCC25 cells by attenuating autophagy. Thus, our findings produced a potential novel agent for the treatment of human OSCC metastasis.

PEI imilti-coated PSBF (PEI-M-PSBF) wth high adsorption efficiency and good reusability for Pd(II) recovery was prepared by a PEI multi-coating method. The results of FT-IR and FE-SEM showed that the PEI-M-PSBF was successfully prepared. Batch experiments were conducted to study the adsorption/desorption performance of PEI-M-PSBF to Pd(II). The Langmuir and pseudo-second-order models were well suited for the batch experiment isotherm and kinetic data, respectively. The adsorption capacity of Pd(II) by Langmuir model was 327.14 mg/g, and the initial adsorption rate of 3-PEI-PSBF was 37.19mg/g min. As a resut, the increase in amine groups on the PEI-M-PSBF surface through the PEI multi-coating directly influenced the adsorption amount of the biosorbent. The effects of initial concentration, bed column depth, and flow rate were determined in a column system and the breakthrough curves were detected by Thomas model. In addition, the recycle of the adsorbent was examined by multiple time performing adsorption/desorption cycles up to ten times in a fixed-bed column.

Highly conductive and stable anion exchange membranes (AEMs) are important components of high-performance anion exchange membrane fuel cells (AEMFCs). Here, we report the use of crosslinked poly(arylene ether sulfone) (PAES) AEMs containing quaternary ammonium (QA) and triazolium cations. The crosslinked PAES-triazole-hydroxide membrane (cPAES-TA-OH) had a higher ion exchange capacity (IEC) than that of the non-crosslinked membrane (PAES-TA-OH) owing to the presence of triazolium cations. The IEC values of cPAES-TA-OH and PAES-TA-OH were 1.75 and 1.31 meq/g, respectively. The IEC value affects the water uptake and swelling ratio of a membrane. The water uptake and swelling ratio of cPAES-TA-OH were higher than those of PAES-TA-OH at 30 °C and 80 °C. In addition, hydroxide conductivity and membrane stability were enhanced by crosslinking; the hydroxide conductivity of cPAES-TA-OH was 92.1 mS/cm at 80 °C under 95% RH (in contrast to 86.2 mS/cm for PAES-TA-OH), and its conductivity retention was 67% after treating with 1M NaOH at 80 °C for 24 h (in contrast to 51% for PAES-TA-OH).

The aim of this study was to investigate the anti-oxidant and anti-inflammatory activities of the Rhododendron weyrichii flower extract fermented using Shindari, a traditional Jeju barley Nuruk-based fermentation. In this study, we examined the antioxidant potential of R. weyrichii flower extracts (RF) and R. weyrichii flower extracts fermented with Nuruk or Shindari (RFFN or RFFS, respectively) using various in vitro antioxidant assays including DPPH and ABTS radical scavenging assays, total phenol content and FRAP assays. We also evaluated the anti-inflammatory activity of the RF and RFFS on murine RAW 264.7 cells. The anti-inflammatory activity was evaluated by treating the RAW 264.7 cells with various concentrations (6.25, 12.5, 25, and 50 μg/ml) of RF or RFFS. As a result, we observed that the ABTS radical scavenging activity and total phenol content of RFFS was higher than that of RF and RFFN. Additionally, lipopolysaccharide- induced nitric oxide (NO) production was significantly lower in RFFS-treated cells when compared to the LPS-treated control. In addition, RFFS-treated cells exhibited decreased expression of inducible NO synthase (iNOS) proteins and high-performance liquid chromatography (HPLC) fingerprinting showed that both the quercetin and quercetin glucoside (quercitrin and isoquercitrin) levels were affected by the fermentation process. In conclusion, our data suggests that traditional fermentation could be an important strategy in improving the biological properties of raw materials including their antioxidant and antiinflammatory activities. Finally, RFFS may be a candidate for developing topical antioxidant and antiinflammatory agents.

Aldose reductase (AR) is known to detoxify aldehydes and prevent oxidative stress. Although AR exerts antioxidant effects, the role of AR in Parkinson's disease (PD) remains unclear. The objective of the present study was to investigate the protective effects of AR protein against 1‑methyl‑4‑phenylpyridinium (MPP+)‑induced SH‑SY5Y cell death and 1‑methyl‑4‑phenyl‑1,2,3,6‑tetrahydropyridine (MPTP)‑induced PD in a mouse model using the cell permeable Tat‑AR fusion protein. The results revealed that when Tat‑AR protein was transduced into SH‑SY5Y cells, it markedly protected the cells against MPP+‑induced death and DNA fragmentation. It also reduced the activation of mitogen-activated protein kinase (MAPKs) and regulated the expression levels of Bcl‑2, Bax and caspase‑3. Immunohistochemical analysis revealed that when Tat‑AR protein was transduced into the substantia nigra (SN) of mice with PD, it markedly inhibited dopaminergic neuronal cell death. Therefore, Tat‑AR may be useful as a therapeutic protein for PD.

A redox-neutral C2-selective methylation of heterocyclic N-oxides with sulfonium ylides is described herein. This report presents unprecedented findings for the utility of sulfonium ylides as the methylation source of N-heterocycles beyond the Corey-Chaykovsky reaction. Intriguingly, pyrrolidine plays a significant role in minimizing the reductive C2-methylation process. This method is characterized by its mild conditions, simplicity, and excellent site selectivity. The applicability of the developed protocol is showcased by the late-stage methylation and sequential transformations of complex drug molecules.

Neovascularization in the retina is common pathophysiology of diabetic retinal microvasculopathy and exudative macular degeneration. Our study assessed the inhibitory activity of an ethanol‐based extract of Aucuba japonica (AJE) on abnormal angiogenesis in the retina with a hyperoxia‐induced neovascular retinopathy model. The inhibitory effects of aucubin, quercetin, and kaempferol, bioactive compounds, from A. japonica, on retinal vascular hyperpermeability were also examined. On the 7th postnatal day (P7), the C57BL/6 pups were exposed to a hyperoxic environment with 75% oxygen to develop the experimental angiogenesis in retinas. On the 12th postnatal day (P12), the pups were then returned to the normal atmospheric pressure of oxygen. From P12 to P16, the administration was intraperitoneal. The dose per day was 250 mg per kg weight. Retinal neovascularization was measured with retinal flat mounts prepared on P17. We also measured the vascular leakage mediated by the vascular endothelial growth factor (VEGF) in retinas. Mice treated with AJE had markedly smaller neovascular lesions, in comparison with vehicle‐administered mice. AJE downregulated the expression of both VEGF protein and mRNA. In addition, aucubin, quercetin, and kaempferol ameliorated VEGF‐induced retinal vascular leakage. The results of our study suggest that AJE is a potent antiangiogenic substance. AJE could also serve as a therapeutic agent for abnormal growth of vessels in the retina in patients with ischemic retinopathy. The bioactive compounds of AJE may be responsible for its antiangiogenic abilities., Aucuba japonica extract (AJE) and its bioactive compounds, aucubin, quercetin, and kaempferol, showed potent anti‐angiogenic activities in a mouse model for oxygen‐induced retinopathy. This study proposes AJE as an anti‐angiogenic functional food for patients with abnormal retinal vessel growth.

Recently, Rodgersia podophylla has been reported to exhibit anti-inflammatory activity. However, little is known about the potential mechanisms about its anti-inflammatory activity. We elucidated the anti-inflammatory mechanisms of leaves extracts from Rodgersia podophylla (RP-L) in RAW264.7 cells. LPS-induced NO was measured by Griess and mRNA of pro-inflammatory mediators was analyzed by RT-PCR. Cell viability was measured using MTT assay. The protein level was analyzed by Western blot. RP-L significantly inhibited the production of the pro-inflammatory mediators such as NO, iNOS, IL-1β and IL-6 in LPS-stimulated RAW264.7 cells. RP-L increased HO-1 expression in RAW264.7 cells, and the inhibition of HO-1 by ZnPP reduced the inhibitory effect of RP-L against LPS-induced NO production in RAW264.7 cells. Inhibition of p38, ROS and GSK3β attenuated RP-L-mediated HO-1 expression. Inhibition of ROS inhibited p38 phosphorylation and GSK3β expression induced by RP-L. In addition, inhibition of GSK3β blocked RP-L-mediated p38 phosphorylation. RP-L induced nuclear accumulation of Nrf2, and inhibition of p38, ROS and GSK3β abolished RP-L-mediated nuclear accumulation of Nrf2. Furthermore, RP-L blocked LPS-induced degradation of IκB-α and nuclear accumulation of p65. RP-L also attenuated LPS-induced phosphorylation of ERK1/2 and p38. In GC/MS analysis of RP-L, pyrogallol was detected as bioactive compound for anti-inflammatory activity of RP-L. Pyrogallol was observed to activate HO-1 expression through ROS/GSK3β/p38/Nrf2/HO-1 signaling. Our results suggest that RP-L exerts potential anti-inflammatory activity by activating ROS/GSK3β/p38/Nrf2/HO-1 signaling and inhibiting NF-κB and MAPK signaling in RAW264.7 cells. These findings suggest that RP-L may have great potential for the development of anti-inflammatory drug.

This study aimed to assess the incidence and distribution of toxigenic fungi in Korean oat. Toxigenic fungi were isolated from oat samples collected from 12 oat fields from heading to harvest in 2017 and 2018. A total of 745 fungal colonies were isolated based on morphology and identified using marker genes. About 92% of the fungal isolates were Fusarium spp. and others were Penicillium (5.9%) and Aspergillus (2.1%). Fusarium isolates comprised mostly of F. asiaticum (83.1%), followed by F. incarnatum (5.4%), F. proliferatum (3.5%), F. fujikuroi (2.8%), F. tricinctum species complex (FTSC) 11 (1.5%) and F. graminearum (1.0%). About 97% of F. asiaticum was nivalenol type, and 3-acetyl deoxynivalenol (3.2%) and 15-acetyl deoxynivalenol (0.4%) types also were found. Pathogenicity test of the selected Fusarium isolates revealed that F. asiaticum isolates have a wide range of virulence depending on the tested plants. F. graminearum and FTSC 11 isolates from blighted spikelets were the most virulent in naked oat. All Fusarium isolates (n=18) except one (FTSC 11) produced nivalenol (0.2–7.6 μg/g), deoxynivalenol (0.03–6.1 μg/g), and zearalenone (0.1–27.0 μg/g) on rice medium. This study is first report that F. asiaticum causes Fusarium head blight disease of oat in Korea. These findings demonstrate the dominance of F. asiaticum in oat agroecosystems as in rice, wheat and barley in Korea.

Olfactory receptors (ORs), which belong to the G-protein-coupled receptor family, have been widely studied as ectopically expressed receptors in various human tissues, including the skin. However, the physiological functions of only a few OR types have been elucidated in skin cells. All-trans retinoic acid (ATRA) is a well-known medication for various skin diseases. However, many studies have shown that ATRA can have adverse effects, resulting from the suppression of cell proliferation. Here, we investigated the involvement of OR7A17 in the ATRA-induced suppression of human keratinocyte (HaCaT) proliferation. We demonstrated that OR7A17 is expressed in HaCaT keratinocytes, and its expression was downregulated by ATRA. The ATRA-induced downregulation of OR7A17 was attenuated via RAR α or RAR γ antagonist treatment, indicating that the effects of ATRA on OR7A17 expression were mediated through nuclear retinoic acid receptor signaling. Moreover, we found that the overexpression of OR7A17 induced the proliferation of HaCaT cells while counteracting the antiproliferative effect of ATRA. Mechanistically, OR7A17 overexpression reversed the ATRA-induced attenuation of Ca2+ entry. Our findings indicated that ATRA suppresses cell proliferation through the downregulation of OR7A17 via RAR α- and γ-mediated retinoid signaling. Taken together, OR7A17 is a potential therapeutic target for ameliorating the anti-proliferative effects of ATRA.

Cancer-associated thrombosis is the second-leading cause of mortality in patients with cancer and presents a poor prognosis, with a lack of effective treatment strategies. NAD(P)H quinone oxidoreductase 1 (NQO1) increases the cellular nicotinamide adenine dinucleotide (NAD+) levels by accelerating the oxidation of NADH to NAD+, thus playing important roles in cellular homeostasis, energy metabolism, and inflammatory responses. Using a murine orthotopic 4T1 breast cancer model, in which multiple thrombi are generated in the lungs at the late stage of cancer development, we investigated the effects of regulating the cellular NAD+ levels on cancer-associated thrombosis. In this study, we show that dunnione (a strong substrate of NQO1) attenuates the prothrombotic state and lung thrombosis in tumor-bearing mice by inhibiting the expression of tissue factor and formation of neutrophil extracellular traps (NETs). Dunnione increases the cellular NAD+ levels in lung tissues of tumor-bearing mice to restore the declining sirtuin 1 (SIRT1) activity, thus deacetylating nuclear factor-kappa B (NF-κB) and preventing the overexpression of tissue factor in bronchial epithelial and vascular endothelial cells. In addition, we demonstrated that dunnione abolishes the ability of neutrophils to generate NETs by suppressing histone acetylation and NADPH oxidase (NOX) activity. Overall, our results reveal that the regulation of cellular NAD+ levels by pharmacological agents may inhibit pulmonary embolism in tumor-bearing mice, which may potentially be used as a viable therapeutic approach for the treatment of cancer-associated thrombosis.

Genome editing took a dramatic turn with the development of the clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated proteins (Cas) system. The CRISPR-Cas system is functionally divided into classes 1 and 2 according to the composition of the effector genes. Class 2 consists of a single effector nuclease, and routine practice of genome editing has been achieved by the development of the Class 2 CRISPR-Cas system, which includes the type II, V, and VI CRISPR-Cas systems. Types II and V can be used for DNA editing, while type VI is employed for RNA editing. CRISPR techniques induce both qualitative and quantitative alterations in gene expression via the double-stranded breakage (DSB) repair pathway, base editing, transposase-dependent DNA integration, and gene regulation using the CRISPR-dCas or type VI CRISPR system. Despite significant technical improvements, technical challenges should be further addressed, including insufficient indel and HDR efficiency, off-target activity, the large size of Cas, PAM restrictions, and immune responses. If sophisticatedly refined, CRISPR technology will harness the process of DNA rewriting, which has potential applications in therapeutics, diagnostics, and biotechnology., Genome editing: Re-writing DNA Yong-Sam Kim and colleagues at Korea Research Institute of Bioscience and Biotechnology in Daejeon, South Korea, outline the latest developments in genome editing technologies and the limitations that remain. Although several approaches have been developed to make changes to an organism’s DNA at particular locations in the genome, the efficiency and simplicity of the CRISPR-Cas system, which plays a defense role in bacteria or archaea by chopping up the invaded DNA, has revolutionized the field. The authors review the different types of CRISPR system and explain how they induce quantitative or qualitative changes in gene expression in eukaryotic cells. Further improving the system’s DNA modification efficiency and the delivery of its components into cells, and finding ways to reduce off-target effects, will be required for CRISPR to realize its therapeutic potential.

Background Vaccinium oldhamii (V. oldhamii) has been reported to exert a variety of the pharmacological properties such as anti-oxidant activity, anti-cancer activity, and inhibitory activity of α-amylase and acetylcholinesterase. However, the anti-inflammatory activity of V. oldhamii has not been studied. In this study, we aimed to investigate anti-inflammatory activity of the stem extracts from V. oldhamii, and to elucidate the potential mechanisms in LPS-stimulated RAW264.7 cells. Methods Cell viability was evaluated by MTT assay. The determination of NO and PGE2 production was performed using Griess reagent and Prostaglandin E2 ELISA Kit, respectively. The change of mRNA or protein level was evaluated by RT-PCR and Western blot. Results Among VOS, VOL and VOF, the inhibitory effect of NO and PGE2 production induced by LPS was highest in VOS treatment. Thus, VOS was selected for the further study. VOS dose-dependently blocked LPS-induced NO and PGE2 production by inhibiting iNOS and COX-2 expression, respectively. VOS inhibited the expression of pro-inflammatory cytokines such as IL-1β, IL-6 and TNF-α. In addition, VOS suppressed TRAP activity and attenuated the expression of the osteoclast-specific genes such as NFATc1, c-FOS, TRAP, MMP-9, cathepsin K, CA2, OSCAR and ATPv06d2. VOS inhibited LPS-induced NF-κB signaling activation through blocking IκB-α degradation and p65 nuclear accumulation. VOS inhibited MAPK signaling activation by attenuating the phosphorylation of ERK1/2, p38 and JNK. Furthermore, VOS inhibited ATF2 phosphorylation and blocked ATF2 nuclear accumulation. Conclusions These results indicate that VOS may exert anti-inflammatory activity by inhibiting NF-κB and MAPK/ATF2 signaling. From these findings, VOS has potential to be a candidate for the development of chemopreventive or therapeutic agents for the inflammatory diseases.

Background/aims Angiotensin II (Ang II) induces podocyte injury resulting in apoptosis in vitro and in vivo. However, the relationship between autophagy and apoptosis in Ang II-induced podocyte injury is unknown and the role of Ang II-induced autophagy in podocyte survival or death remains unclear. We investigated the sequential relationship between autophagy and apoptosis in Ang II-induced podocytes as well as the role of phosphatidylinositide 3-kinase (PI3-kinase). Methods Mouse podocytes were incubated in media containing various concentrations of Ang II and at different incubation times. The changes of podocyte autophagy and apoptosis were observed by electron microscopy, confocal imaging, western blotting, and FACS assay according to the presence of Ang II. Results Ang II enhanced the podocyte expression of the autophagic proteins, LC3A/B-II and beclin-1, and also increased the number of autophagosomes compared with control cells at early phase of 12 hours in a dose-dependent manner. This effect was inhibited by pretreatment with 3-methyladenine (3-MA), a PI3-kinase class III inhibitor. Thereafter, the Ang II-induced enhancement in autophagy decreased, whereas, podocyte apoptosis appeared later at 24 hours in concentration- and time-dependent manners in FACS and TUNEL assays. 3-MA and LY294002, a pan PI3-kinase inhibitor, further increased Ang II-induced podocyte apoptosis. Suppression of autophagy by Atg5 siRNA could induce podocyte apoptosis and further augment high-dose Ang II-induced podocyte apoptosis. Conclusion These findings suggest that Ang II promotes autophagy in podocytes before apoptosis as an early adaptive cytoprotective mechanism for podocyte survival after Ang II treatment, and the transitional imbalance between autophagy and apoptosis causes podocyte injury.

Piperlongumine (PL) is a natural product found in long pepper (Piper longum ). The pharmacological effects of PL are well known, and it has been used for pain, hepatoprotection, and asthma in Oriental medicine. No studies have examined the effects of PL on bone tissue or bone-related diseases, including osteoporosis. The current study investigated for the first time the inhibitory effects of PL on osteoclast differentiation, bone resorption, and osteoclastogenesis-related factors in RAW264.7 macrophages stimulated by the receptor activator for nuclear factor- κB ligand (RANKL). Cytotoxicity was examined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, and osteoclast differentiation and bone resorption were confirmed by tartrate-resistant acid phosphatase (TRAP) staining and pit formation analysis. Osteoclast differentiation factors were confirmed by western blotting. PL exhibited toxicity in RAW264.7 macrophages, inhibiting osteoclast formation and bone resorption, in addition to inhibiting the expression of osteoclastogenesis-related factors, such as tumor necrosis factor receptor-associated factor 6 (TRAF6), c-Fos, and NFATc1, in RANKL-stimulated RAW264.7 macrophages. These findings suggest that PL is suitable for the treatment of osteoporosis, and it serves as a potential therapeutic agent for various bone diseases.

To characterize the immuno-stimulating ingredient from the Korean citrus, Cheongkyool, a crude polysaccharide (CCE-0) was isolated from the pectinase digests of Cheongkyool peels, from which the complex polysaccharide CCE-I was purified to homogeneity by gel filtration. CCE-I highly enhanced the production of IL-6, TNF-α, and NO in RAW 264.7 cell lines. It augmented the mRNA expression of IL-6, TNF-α, and iNOS in a dose-dependent manner. Moreover, CCE-I dose-dependently induced phosphorylation of MAPKs and NF-κB related proteins and led to the nuclear translocation of p65. The effect of CCE-I on NO and IL-6 production was suppressed by treatment with specific antibodies for TLR2, TLR4, and scavenger receptors. Conversely, the primary structure of CCE-I that exhibited potent immunostimulatory activity was characterized by sugar composition, linkage analysis, and oligosaccharide analysis after β-elimination. The results suggested that CCE-I may be a rhamnogalacturonan-I type, highly branched polysaccharide with short arabinan and galactan side chains.

A chitin-based biosorbent that can be used to effectively remove dyes, typical color inducers in industrial wastewater, was developed. The crosslinking agent glutaraldehyde was bonded to the hydroxyl and/or amine groups of chitin, and then polyethylenimine (PEI), a cationic polymer, was cross-linked to produce PEI-chitin. The successful preparation of PEI-chitin was confirmed using FE-SEM, BET and FTIR analysis and simple adsorption evaluation. The adsorption performance of PEI-chitin was assessed by Acid Blue 25, in terms of the pH effect, isotherm adsorption, adsorption kinetic and thermodynamic parameters. Based on the Langmuir model the maximum dye uptake at pH 2 was 177.32 mg/g, and the adsorption equilibrium was attained within 120 and 500 min for two different dye concentrations (50 and 100 mg/L). Desorption experiments were evaluated using 0.001–1 M NaOH, NaHCO3, and Na2CO3 as eluents. The desorption rate was highest (85.2%) when 0.01 M NaOH was employed as the eluent. It was found that the developed biosorbent could be repeatedly used three times through adsorption-desorption of Acid Blue 25.

GS-E3D is an enzymatically modified ginseng extract by pectin lyase. In this study, we evaluated the preventive effects of GS-E3D on blood-retinal barrier (BRB) leakage in a rat model of diabetes. To produce diabetes, rats were injected with streptozotocin. GS-E3D was orally gavaged at 25, 50, and 100 mg/kg body weight for 6 weeks. We then compared the effect of GS-E3D with that of an unmodified ginseng extract (UGE) on retinal vascular leakage. The administration of GS-E3D significantly blocked diabetes-induced BRB breakdown. Immunofluorescence staining showed that GS-E3D reduced the loss of occludin in diabetic rats. In TUNEL staining, the number of apoptotic retinal microvascular cells was dose dependently decreased by GS-E3D treatment. GS-E3D decreased the accumulations of advanced glycation end products in the retinal vessels. In addition, the inhibition potential of GS-E3D on BRB breakage was stronger compared with UGE. These results indicate that GS-E3D could be a beneficial treatment option for preventing diabetes-induced retinal vascular injury.

Development of a thin and dry electrode for electrocardiography (ECG) is essential in order to prevent skin irritation, allergic reactions from electrolytic gel, and motion artifacts caused by relative motion between the electrodes and the skin. In this study, we have developed a composite electrode made from Ag nanowires (AgNWs) and polyvinyl butyral (PVB), prepared by inverted layer processing (ILP). The initial composite electrodes were mechanically stable, flexible, and transparent; however, most of the NWs were located beneath the surface of the PVB such that few conductive pathways were exposed and available to contact the skin. In order to resolve this issue, prior to transferring the AgNWs from the temporary glass substrate to the PVB, we irradiated the NWs with intensive pulsed light. This irradiation induced plasmonic heating of the AgNWs, which caused the NWs to sink towards the glass and form a dense layer on the temporary substrate. Subsequent ILP resulted in the fabrication of an AgNWs/PVB composite electrode that demonstrated significant surface coverage of conductive pathways available for stable electrical contact with skin. The resultant composite electrode is an improved ECG electrode that exhibits fewer motion artifacts compared to conventional Ag/AgCl-based wet electrodes since it is both dry and conformable.

For next generation Li-ion batteries, advanced electrode materials with high energy densities are mightily important issue. Since the partial oxidation of Sn with Li2O to form SnOx was demonstrated upon delithiation in SnO2 anode, the additional conversion reaction has contributed that the theoretical capacity of SnO2 can be extended from 783 to 1494 mAh g−1. Herein, with the design of additional conversion reaction, we discuss key factors for high electrochemical performances of the SnO2 anodes through comparative analysis between nano-structured mesoporous SnO2 and conventional bulk SnO2, based on synchrotron radiation-based techniques, quantitative analysis of extended X-ray absorption fine structure spectra, and bond strength calculation. In this way, we demonstrate that the mesoporous SnO2 has a nano-engineered structure with the weakened Sn-O bond strength for inducing the enhanced reversible conversion reaction as well as the facilitated electrochemical reaction, and a void structure for relieving the severe volume changes during lithiation/delithiation. Consequently, excellent electrochemical performance through the additional reversible conversion reaction is obtained in the mesoporous SnO2. Insight from this research enables important advance in the development of metal oxide-based anode materials by making irreversible reaction reversible, providing a realizable strategy for the design and creation of high-energy storage devices.

Background Sageretia thea (S. thea) has been used as the medicinal plant for treating hepatitis and fevers in Korea and China. Recently, anticancer activity of S. thea has been reported, but the potential mechanism for the anti-cancer property of S. thea is still insufficient. Thus, we evaluated whether extracts from the leaves (STL) and branches (STB) of S. thea exert anticancer activity and elucidated its potential mechanism in SW480 cells. Methods MTT assay was performed for measuring cell viability. Western blot and RT-PCR were used for analyzing the level of protein and mRNA, respectively. Results Treatment of STL or STB decreased the cell viability and induced apoptosis in SW480 cells. Decreased level of cyclin D1 protein was observed in SW480 cells treated with STL or STB, but no change in cyclin D1 mRNA level was observed with the treatment of STL or STB. MG132 blocked downregulation of cyclin D1 protein by STL or STB. Thr286 phosphorylation of cyclin D1 by STL or STB occurred faster than downregulation of cyclin D1 protein in SW480 cells. When SW480 cells were transfected with T286A-cyclin D1, cyclin D1 degradation by STL or STB did not occur. Inhibition of GSK3β and cyclin D1 nuclear export attenuated STL or STB-mediated cyclin D1 degradation. In addition, STL or STB increased HO-1 expression, and the inhibition of HO-1 attenuated the induction of apoptosis by STL or STB. HO-1 expression by STL or STB resulted from Nrf2 activation through ROS-dependent p38 activation. Conclusions These results indicate that STL or STB may induce GSK3β-dependent cyclin D1 degradation, and increase HO-1 expression through activating Nrf2 via ROS-dependent p38 activation, which resulted in the decrease of the viability in SW480 cells. These findings suggest that STL or STB may have great potential for the development of anti-cancer drug.

Sageretia thea (S. thea) commonly known as Chinese sweet plum or Chinese bird plum has been used for treating hepatitis and fevers in Korea and China. S. thea has been reported to exert anti-oxidant, anticancer and anti-human immunodeficiency virus activity. However, there is little study on the anti-inflammatory activity of S. thea. Thus, we evaluated the anti-inflammatory effect of extracts of leaves (ST-L) and branches (ST-B) from Sageretia thea in LPS-stimulated RAW264.7 cells. ST-L and ST-B significantly inhibited the production of the pro-inflammatory mediators such as NO, iNOS, COX-2, IL-1[Formula: see text] and IL-6 in LPS-stimulated RAW264.7 cells. ST-L and ST-B blocked LPS-induced degradation of I[Formula: see text]B-[Formula: see text] and nuclear accumulation of p65, which resulted in the inhibition of NF-[Formula: see text]B activation in RAW264.7 cells. ST-L and ST-B also attenuated the phosphorylation of ERK1/2, p38 and JNK in LPS-stimulated RAW264.7 cells. In addition, ST-L and ST-B increased HO-1 expression in RAW264.7 cells, and the inhibition of HO-1 by ZnPP reduced the inhibitory effect of ST-L and ST-B against LPS-induced NO production in RAW264.7 cells. Inhibition of p38 activation and ROS elimination attenuated HO-1 expression by ST-L and ST-B, and ROS elimination inhibited p38 activation induced by ST-L and ST-B. ST-L and ST-B dramatically induced nuclear accumulation of Nrf2, but this was significantly reversed by the inhibition of p38 activation and ROS elimination. Collectively, our results suggest that ST-L and ST-B exerts potential anti-inflammatory activity by suppressing NF-[Formula: see text]B and MAPK signaling activation, and activating HO-1 expression through the nuclear accumulation of Nrf2 via ROS-dependent p38 activation. These findings suggest that ST-L and ST-B may have great potential for the development of anti-inflammatory drug to treat acute and chronic inflammatory disorders.

Fisetin (3,3-,4-,7-tetrahydroxyflavone), a naturally occurring flavonoid, has antioxidant, anti-inflammatory, and anticancer effects. Oral squamous cell carcinoma (OSCC) has a 5-year survival rate lower than that of most other carcinomas, and can create functional and aesthetic problems for the patient. New therapies for OSCC are necessary, and treatment using plant-derived natural substances has recently become a trend. It has been suggested that autophagy may play an important role in cancer therapy. Several studies demonstrated that autophagy inhibition enhances apoptotic cell death. Therefore, autophagy inhibition might be a promising therapeutic method against OSCC. Our results showed that fisetin induced apoptotic cell death in human tongue squamous cell line Ca9-22 could be enhanced by inhibition of autophagy. Thus, autophagy process in fisetin treated OSCC might presumed to play a role of pro-survival. The combination of fisetin and an effective autophagy inhibitor could be a potentially adjuvant and useful treatment for oral cancer.

A new electron-rich central core (BTT) based on benzo[1,2-b:4,5-b′]dithiophene with a thio[3,2-b]thiophene side chain was coupled with methyl and fluorine-modified 1,1-dicyanomethylene-3-indanone (IC) electron-withdrawing end groups to furnish two novel non-fullerene acceptors (NFAs), namely BTT-MIC and BTT-FIC. Compared to BTT-MIC, BTT-FIC exhibits a stronger near-infrared (NIR) absorption with a smaller optical bandgap (1.47 eV). The BTT-FIC/PM6 based device achieved a champion PCE of 12.65% for benzo[1,2-b:4,5-b′]di(cyclopenta[2,1-b:3,4-b′]dithiophene)-containing NFA based organic solar cells (OSCs), while the BTT-FIC/PM6 based device delivered a PCE of 10.04% with a high Voc of over 1 V. Fluorinated IC is more suitable for this new core BTT than methyl substituted IC, because fluorine atoms enhance the charge transport (CT) state (absorption) and are also beneficial to exciton dissociation, charge mobility and morphology. This work revealed that BTT with a thieno[3,2-b]thiophene side chain is an effective electron-donating core for creating effective NFAs and fluorinated IC is a suitable end group for BTT.

This study reports the recovery of Pd(II) from acid solution by a polyethylenimine (PEI)-crosslinked chitin (PEI-chitin) biosorbent. FE-SEM analysis demonstrated that there are many slot-like pores on PEI-chitin. The N2 adsorption–desorption experiment revealed that the average pore size was 47.12 nm. Elemental analysis verified the successful crosslinking of PEI with raw chitin. The Langmuir model better explained the isotherm experimental data and the theoretical maximum Pd(II) uptake was 57.1 mg/g. The adsorption kinetic data were better described by the pseudo-second-order model and the adsorption equilibrium was achieved within 30 min for all initial Pd(II) concentrations of 50–200 mg/L. In the fixed-bed column, the adsorption of Pd(II) on PEI-chitin showed a slow breakthrough and a fast saturation performance. The desorption experiments achieved a concentration factor of 8.4 ± 0.4, in addition, the adsorption–desorption cycles in the fixed-bed column were performed up to three times, consequently confirming the good reusability of PEI-chitin for Pd(II) recovery. Therefore, the PEI-chitin can be used as a promising biosorbent for the recovery of Pd(II) in practical applications.

Mixed metal oxide (MMO) is one of the widely utilized ceramic materials in various industries. In order to obtain high performance, the specific surface area of MMO should be controlled. Calcination of layered double hydroxide (LDH) is a versatile way to prepare MMO with homogeneous metal distribution and well-developed porosity. Although researchers found that the specific surface area of LDH-originated MMO was relatively high, it had not been systematically investigated how the surface area is controlled under a certain parameter. In this review, we summarized LDH-originated MMO with various starting composition, calcination temperature, and pore developing agent in terms of specific surface area and porosity. Briefly, it was represented that MMOs with Mg-Al components generally had higher specific surface area than Mg-Fe or Zn-Al components. Calcination temperature in the range 300–600 °C resulted in the high specific surface area, while upper or lower temperature reduced the values. Pore developing agent did not result in dramatic increase in MMO; however, the pore size distribution became narrower in the presence of pore developing agents.

Background Nannochloropsis is a marine microalga that has been extensively studied. The major carotenoid produced by this group of microalgae is violaxanthin, which exhibits anti-inflammatory, anti-photoaging, and antiproliferative activities. Therefore, it has a wide range of potential applications. However, large-scale production of this pigment has not been much studied, thereby limiting its industrial application. Results To develop a novel strain producing high amount of violaxanthin, various Nannochloropsis species were isolated from seawater samples and their violaxanthin production potential were compared. Of the strains tested, N. oceanica WS-1 exhibited the highest violaxanthin productivity; to further enhance the violaxanthin yield of WS-1, we performed gamma-ray-mediated random mutagenesis followed by colorimetric screening. As a result, Mutant M1 was selected because of its significant higher violaxanthin content and biomass productivity than WS-1 (5.21 ± 0.33 mg g− 1 and 0.2101 g L− 1 d− 1, respectively). Subsequently, we employed a 10 L-scale bioreactor to confirm the large-scale production potential of M1, and the results indicated a 43.54 % increase in violaxanthin production compared with WS-1. In addition, comparative transcriptomic analysis performed under normal light condition identified possible mechanisms associated with remediating photo-inhibitory damage and other key responses in M1, which seemed to at least partially explain enhanced violaxanthin content and delayed growth. Conclusions Nannochloropsis oceanica mutant (M1) with enhanced violaxanthin content was developed and its physiological characteristics were investigated. In addition, enhanced production of violaxanthin was demonstrated in the large-scale cultivation. Key transcriptomic responses that are seemingly associated with different physiological responses of M1 were elucidated under normal light condition, the details of which would guide ongoing efforts to further maximize the industrial potential of violaxanthin producing strains.

In this study, we investigated the microbial community and its correlation with anti-inflammatory effects during the fermentation procedure of shindari. Since shindari is traditionally manufactured through a fermentation process of 3–4 days in summer or 5–6 days in winter, the shindari materials applied in this study were fermented for 1, 3, and 5 days, respectively. Microbial community structure in the shindari manufacturing process was analyzed by Illumina MiSeq sequencing. Results revealed different fermentation time had different influences on the community structure and microbial diversity in the shindari process. Ultimately, we found that pathogenic bacteria such as Enterococcus durans (9.3%) and Enterobacter asburiae (4.5%) dominate the shindari fermented for 1 day, while samples with a fermentation period of more than 3 days occupied lactic acid bacteria such as Pediococcus sp. (97.1%). In addition, to study the relationship between fermentation period and anti-inflammatory effect in the shindari manufacturing process, we applied RAW264.7 cells, a classic cell model for inflammation studies. First, we prepared an ethyl acetate extract of shindari fermented for 1 (S1), 3 (S3), or 5 days (S5), and then it was confirmed that all of these extracts inhibited the nitric oxide (NO) production in a concentration-dependent manner. In addition, these inhibitory effects were correlated with the suppressive effect of shindari extracts against overexpression of inducible nitric oxide synthase (iNOS). Furthermore, S3 and S5 also inhibited the production of pro-inflammatory cytokines such as tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and IL-6. Overall, the anti-inflammatory effect of S3 was suggested to be mediated through the negative regulation of mitogen-activated protein kinase signaling (MAPKs) and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) signaling pathways.

Cryptococcus neoformans is a human-pathogenic fungal pathogen that causes life-threatening meningoencephalitis in immunocompromised individuals. To investigate the roles of N-glycan core structure in cryptococcal pathogenicity, we constructed mutant strains of C. neoformans with defects in the assembly of lipid-linked N-glycans in the luminal side of the endoplasmic reticulum (ER). Deletion of ALG3 (alg3Δ), which encodes dolichyl-phosphate-mannose (Dol-P-Man)-dependent α-1,3-mannosyltransferase, resulted in the production of truncated neutral N-glycans carrying five mannose residues as a major species. Despite moderate or nondetectable defects in virulence-associated phenotypes in vitro, the alg3Δ mutant was avirulent in a mouse model of systemic cryptococcosis. Notably, the mutant did not show defects in early stages of host cell interaction during infection, including attachment to lung epithelial cells, opsonic/nonopsonic phagocytosis, and manipulation of phagosome acidification. However, the ability to drive macrophage cell death was greatly decreased in this mutant, without loss of cell wall remodeling capacity. Furthermore, deletion of ALG9 and ALG12, encoding Dol-P-Man-dependent α-1,2-mannosyltransferases and α-1,6-mannosyltransferases, generating truncated core N-glycans with six and seven mannose residues, respectively, also displayed remarkably reduced macrophage cell death and in vivo virulence. However, secretion levels of interleukin-1β (IL-1β) were not reduced in the bone marrow-derived dendritic cells obtained from Asc- and Gsdmd-deficient mice infected with the alg3Δ mutant strain, excluding the possibility that pyroptosis is a main host cell death pathway dependent on intact core N-glycans. Our results demonstrated N-glycan structures as a critical feature in modulating death of host cells, which is exploited by as a strategy for host cell escape for dissemination of C. neoformans. IMPORTANCE We previously reported that the outer mannose chains of N-glycans are dispensable for the virulence of C. neoformans, which is in stark contrast to findings for the other human-pathogenic yeast, Candida albicans. Here, we present evidence that an intact core N-glycan structure is required for C. neoformans pathogenicity by systematically analyzing alg3Δ, alg9Δ, and alg12Δ strains that have defects in lipid-linked N-glycan assembly and in in vivo virulence. The alg null mutants producing truncated core N-glycans were defective in inducing host cell death after phagocytosis, which is triggered as a mechanism of pulmonary escape and dissemination of C. neoformans, thus becoming inactive in causing fatal infection. The results clearly demonstrated the critical features of the N-glycan structure in mediating the interaction with host cells during fungal infection. The delineation of the roles of protein glycosylation in fungal pathogenesis not only provides insight into the glycan-based fungal infection mechanism but also will aid in the development of novel antifungal agents.

This study analyzed the antioxidant activity, cell viability, and human skin primary irritation test using the hot-water extracts of the Syzygium samarangense. As a result of the recent warmer climate, tropical plants have flourished on Jeju Island, and S. samarangense is one of these plants known to have biological activities. In this study, the hot-water extract of S. samarangense leaf and branch was analyzed. Antioxidant activity was measured by DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS (2,2&rsquo, azino-bis(3-ethyl-benzthiazoline-6-sulfonic acid)) assays, and the DMPD (dimethyl-4-phenylenediamine) radical scavenging activity, nitrite scavenging activity, ferrous-ion chelating activity, cupric reducing antioxidant capacity, reducing power assay, ferric reducing antioxidant power, total phenol content, and total flavonoid content were also measured. In addition, cell viability was measured by MTT assay in human keratinocyte cells (HaCaT), and the safety of the extract for use on the skin was evaluated in the human skin primary irritation test. The antioxidant activities, except DMPD radical scavenging activity and ferrous-ion chelating activity, were stronger in the branch extract than in leaf extract, and the total phenol and flavonoid contents were also higher in the branch extract. Slight irritation was observed in the human skin primary irritation test. However, it was possible to observe sufficient antioxidant capacity at a concentration lower than the concentration used in the irritation test, therefore, if the concentration of the extract is appropriately adjusted, this suggests that it is a possible natural material suitable for use in cosmetics.

PF-543, the most potent sphingosine kinase (SK) inhibitor, does not demonstrate effective anticancer activity in some cancer cells, unlike other known SK1 inhibitors. PF-543 has a non-lipid structure with a unique toluene backbone, however, the importance of this structure remains unclear. Therefore, the purpose of this study was to investigate changes in SK inhibitory and anticancer activities and to explore the role of the tolyl group structure of PF-543 through various modifications. We transformed the methyl group of PF-543 into hydrogen, fluorine, and hydroxy. PF-543 derivatives in which the methyl group was substituted by hydrogen and fluorine (compound 5) demonstrated SK1 inhibitory and anticancer activities similar to PF-543. Moreover, we performed molecular modeling studies of PF-543 and compound 5. To assess the metabolic stability of PF-543 and compound 5, we determined their degree of degradation using the liver microsomes of four different animal species (human, dog, rat, and mouse). However, both PF-543 and compound 5 showed poor microsomal stability. Therefore, for the medical applications of PF-543, the structural modifications of its other parts may be necessary. Our results provide important information for the design of additional PF-543 analogs.

Background For decades, plastic has been a valuable global product due to its convenience and low price. For example, polyethylene terephthalate (PET) was one of the most popular materials for disposable bottles due to its beneficial properties, namely impact resistance, high clarity, and light weight. Increasing demand of plastic resulted in indiscriminate disposal by consumers, causing severe accumulation of plastic wastes. Because of this, scientists have made great efforts to find a way to biologically treat plastic wastes. As a result, a novel plastic degradation enzyme, PETase, which can hydrolyze PET, was discovered in Ideonella sakaiensis 201-F6 in 2016. Results A green algae, Chlamydomonas reinhardtii, which produces PETase, was developed for this study. Two representative strains (C. reinhardtii CC-124 and CC-503) were examined, and we found that CC-124 could express PETase well. To verify the catalytic activity of PETase produced by C. reinhardtii, cell lysate of the transformant and PET samples were co-incubated at 30 °C for up to 4 weeks. After incubation, terephthalic acid (TPA), i.e. the fully-degraded form of PET, was detected by high performance liquid chromatography analysis. Additionally, morphological changes, such as holes and dents on the surface of PET film, were observed using scanning electron microscopy. Conclusions A PET hydrolyzing enzyme, PETase, was successfully expressed in C. reinhardtii, and its catalytic activity was demonstrated. To the best of our knowledge, this is the first case of PETase expression in green algae.

Background The initial step of influenza infection is binding of the virus to specific sialic acid receptors expressed by host cells. This is followed by cell entry via endocytosis. Cleavage of the influenza virus hemagglutinin (HA) protein is critical for infection; this is performed by host cell proteases during viral replication. In cell culture systems, HA is cleaved by trypsin added to the culture medium. The vast majority of established cell lines are mammalian. Results In the present study, we generated genetically engineered chicken DF-1 cell lines overexpressing transmembrane protease, serine 2 (TMPRSS2, which cleaves HA), ST3 beta-galactoside alpha-2,3-sialyltransferase 1 (ST3GAL1, which plays a role in synthesis of α-2,3 linked sialic acids to which avian-adapted viruses bind preferentially), or both. We found that overexpression of TMPRSS2 supports the virus life cycle by cleaving HA. Furthermore, we found that overexpression of ST3GAL1 increased the viral titer. Finally, we showed that overexpression of both TMPRSS2 and ST3GAL1 increased the final viral titer due to enhanced support of viral replication and prolonged viability of the cells. In addition, overexpression of these genes of interest had no effect on cell proliferation and viability. Conclusions Taken together, the results indicate that these engineered cells could be used as a cell-based system to propagate influenza virus efficiently in the absence of trypsin. Further studies on influenza virus interactions with chicken cell host factors could be studied without the effect of trypsin on cells.

Phosphoprotein enriched in astrocytes 15 (PEA15) plays a multi-functional role in neuronal cell survival, however the effects of PEA15 against inflammation have not been investigated yet. To examine the effects of PEP-1-PEA15 protein against lipopolysaccharide (LPS)-induced inflammatory responses in Raw 264.7 cells and in a 12-O-tetradecanoylphobol 13-acetate (TPA)-induced mouse model, we constructed and purified PEP-1-PEA15 protein, which can transduce into cells or tissues. PEP-1-PEA15 inhibited LPS-induced damage in cells including that caused by reactive oxygen species (ROS) production and DNA fragmentation. PEP-1-PEA15 also significantly suppressed activation of mitogen activated protein kinases (MAPKs), pro-inflammatory mediator proteins and various cytokines. In a TPA-induced mouse ear edema model, PEP-1-PEA15 significantly reduced ear weight and thickness as well as MAPK activation as well as the expression levels of COX-2, iNOS, IL-6, IL-1β, and TNF-α. These results demonstrated that PEP-1-PEA15 showed anti-inflammatory effect in cells and animal model suggesting that this fusion protein protects cells or skin tissues from inflammatory response.

This study aimed to investigate in vitro and in vivo the probiotic characteristics of lactic acid bacteria (LAB) isolated from Korean traditional fermented foods. Caenorhabditis elegans (C. elegans) was used for analytical assays of fertility, chemotaxis, life-span, worm-killing and bacterial colonization in the intestinal lumen of the worm. All 35 strains of LAB reduced fertility and slowed development in the worms. The worm-killing assay showed that LAB significantly increased the lifespan (P

The aim of this study was to examine the ameliorating effect of Spirulina platensis (SP) on cognitive dysfunction in high-fat-diet (HFD) mice. Glucose tolerance was evaluated using the intraperitoneal glucose tolerance test (IPGTT). The results showed that the area under the curve (AUC) decreased 20% in the SP group compared to that in the HFD group. The ameliorating effect on HFD-induced learning and memory impairment was investigated using Y-maze and Morris water maze tests. The administration of SP effectively restored learning and memory function compared to that in the HFD group. Using blood serum analysis, liver and kidney function and lipid metabolism-related biomarkers were measured. Generally, administration of SP effectively restored lipid-metabolism by increasing HDLC (HTR) and decreasing LDLC. The antioxidant effects in tissues were also investigated by measuring the superoxide dismutase (SOD) content, oxidized glutathione (GSH)/total GSH ratio, and malondialdehyde (MDA) content, and the administration of SP effectively improved the antioxidant system in the brain and liver tissues. The improvement of cognitive function was confirmed by the inhibitory effect of acetylcholinesterase (AChE) as a biomarker of the cholinergic system, which plays an important role in neurotransmission, and the SP group showed significant inhibition of AChE. Based on these results, SP could be used to improve brain function by ameliorating the HFD condition.

A novel series of blended polymer electrolyte membranes (PEMs) from crosslinked sulfonated poly(arylene ether) (cSPAE100) and triazole-containing poly(arylene ether sulfone) (PAES-DTM) were prepared for fuel cell applications. cSPAE100 with a high degree of sulfonation was synthesized via aromatic condensation. PAES-DTM densely functionalized with di-triazole moieties was blended with cSPAE100. The blend ratios of the fabricated PEMs varied from 2 wt% to 10 wt%, and the blend PEMs (cSPAE-DTMs) displayed sufficiently high ion exchange capacities (IEC = 2.07–1.86 mequiv/g). Introducing the triazole moieties, the proton conductivities of cSPAE-DTM2 and cSPAE-DTM5 increased at high relative humidity (95% RH), whereas their IEC values decreased. Furthermore, evaluation of the cell performance at 80 °C revealed that the performance of the cSPAE-DTMs was superior to that of the pristine membrane under 50% RH. For practical application in fuel cells, the thermal, chemical and mechanical properties of all membranes were investigated.

Oxidative desulfurization (ODS) of organic compounds containing sulfur element from a model oil was performed using tungsten oxide catalysts supported on mesoporous silica with cubic Ia3d mesostructure, well-defined mesopores (7.2 nm), high surface area (719 m2/g), and three-dimensional pore network (WO x /KIT-6). The prepared WO x /KIT-6 catalysts (5–20 wt% WO x ) were characterized by X-ray diffraction analysis, N2 sorption measurements, electron microscopy, H2-temperature programmed reduction, Raman spectroscopy, and thermogravimetric analysis. Among the mesoporous catalysts, 10 wt% WO x /KIT-6 exhibited the best catalytic performance. Sulfur-containing organic compounds, such as dibenzothiophene, 4,6-dimethyldibenzothiophene, and benzothiophene, were completely (100 %) removed from the model oil over 10 wt% WO x /KIT-6 catalyst in 2 h. In addition, the catalyst could be reused several times with only slight decrease in catalytic activity.

Delphinidin is major anthocyanidin that is extracted from many pigmented fruits and vegetables. This substance has anti‐oxidant, anti‐inflammatory, anti‐angiogenic, and anti‐cancer properties. In addition, delphinidin strongly suppresses the migration and invasion of various cancer cells during tumorigenesis. Although delphinidin has anti‐cancer effects, little is known about its functional roles in osteosarcoma (OS). For these reasons, we have demonstrated the effects of delphinidin on OS cell lines. The effects of delphinidin on cell viability and growth of OS cells were assessed using the MTT assay and colony formation assays. Hoechst staining indicated that the delphinidin‐treated OS cells were undergoing apoptosis. Flow cytometry, confocal microscopy, and a western blot analysis also indicated evidence of apoptosis. Inhibition of cell migration and invasion was found to be associated with epithelial‐to‐mesenchymal transition (EMT), observed by using a wound healing assay, an invasion assay, and a western blot analysis. Furthermore, delphinidin treatment resulted in a profound reduction of phosphorylated forms of ERK and p38. These findings demonstrate that delphinidin treatment suppressed EMT through the mitogen‐activated protein kinase (MAPK) signaling pathway in OS cell lines. Taken together, our results suggest that delphinidin strongly inhibits cell proliferation and induces apoptosis. Delphinidin treatment also suppresses cell migration and prevents EMT via the MAPK‐signaling pathway in OS cell lines. For these reasons, delphinidin has anti‐cancer effects and can suppress metastasis in OS cell lines, and it might be worth using as an OS therapeutic agent.

Ionic liquids are effective additives in terms of improving the key electrolyte properties including the ionic conductivity and the oxidative solubility of Grignard reagent-based electrolytes for rechargeable magnesium batteries. However, a precise understanding of their working mechanisms remains elusive to date. Here, we unravel for the first time the mechanism behind the drastic improvement of key electrolyte properties of Grignard reagents upon the addition of allyl-functionalized pyrrolidinium-based ionic liquids. We show that the Grignard reagents selectively abstract acidic protons in the allyl functional group to create a series of Mg-complexes that are remarkably stabilized by the formation of resonance structures. Moreover, the properties of the resulting electrolytes are tuned by adjusting the molar concentration of the ionic liquids which determines the chemical reaction pathway for the formation of new Mg-complexes. Overall, this study demonstrates a novel strategic approach for developing highly efficient new electrolyte systems for rechargeable magnesium batteries.

Aster koraiensis extract (AKE) is a standard dietary herbal supplement. Chlorogenic acid (CA) is the major compound present in AKE. Retinal neovascularization is a common pathophysiology of retinopathy of prematurity, diabetic retinopathy, and wet form age-related macular degeneration. In this study, we aimed to evaluate the effects of AKE and CA on retinal neovascularization in a mouse model of oxygen-induced retinopathy (OIR). Vascular endothelial growth factor- (VEGF-) induced tube formation was assayed in human vascular endothelial cells. Experimental retinal neovascularization was induced by exposing C57BL/6 mice to 75% oxygen on postnatal day 7 (P7) and then returning them to normal oxygen pressure on P12. AKE (25 and 50 mg/kg/day) and CA (25 and 50 mg/kg/day) were administered intraperitoneally for 5 days (P12–P16). Retinal flat mounts were prepared to measure the extent of retinal neovascularization at P17. The incubation of human vascular endothelial cells with AKE and CA (1–10 μg/mL) resulted in the inhibition of VEGF-mediated tube formation in a dose-dependent manner. The neovascular area was significantly smaller in AKE or CA-treated mice than in the vehicle-treated mice. These results suggest that AKE is a potent antiangiogenic agent and that its antiangiogenic activity may, in part, be attributable to the bioactive component CA.

Artemisia princeps var. orientalis (Asteraceae, A. princeps) is a well-known traditional medicinal herb used for treating various inflammatory disorders in Korea, Japan, China, and other Asian countries. In the present study, we investigated the effects of A. princeps extract (APO) on interleukin- (IL-) 1β regulation and inflammasome activation in bone marrow-derived macrophages (BMDMs) and monosodium urate- (MSU-) induced peritonitis mouse model in vivo. The APO treatment to BMDMs primed with lipopolysaccharide (LPS) attenuated the NLRP3 and AIM2 inflammasome activation induced by danger signals, such as ATP, nigericin, silica crystals, and poly (dA:dT), respectively. Mechanistic study revealed that APO suppressed the ASC oligomerization and speck formation, which are required for inflammasome activation. APO treatment also reduced the ASC phosphorylation induced by the combination of LPS and a tyrosine phosphatase inhibitor. In vivo evaluation revealed that intraperitoneal administration of APO reduced IL-1β levels, significantly (p<0.05) and dose dependently, in the MSU-induced peritonitis mouse model. In conclusion, our study is the first to report that the extract of A. princeps inhibits inflammasome activation through the modulation of ASC phosphorylation. Therefore, APO might be developed as therapeutic potential in the treatment of inflammasome-mediated inflammatory disorders, such as gouty arthritis.