Bioactivity-Guided Fraction from Viscera of Abalone, Haliotis discus hannai Suppresses Cellular Basophils Activation and Anaphylaxis in Mice.

Basophils and mast cells are specialized effector cells in allergic reactions. Haliotis discus hannai (abalone), is valuable seafood. Abalone male viscera, which has a brownish color and has not been previously reported to show anti-allergic activities, was extracted with acetone. Six different acetone/hexane fractions (0, 10, 20, 30, 40, and 100%) were obtained using a silica column via ß-hexosaminidase release inhibitory activity-guided selection in phorbol myristate acetate and a calcium ionophore, A23187 (PMACI)-induced human basophils, KU812F cells. The 40% acetone/hexane fraction (A40) exhibited the strongest inhibition of PMACI-induced-ß-hexosaminidase release. This fraction dose-dependently inhibited reactive oxygen species (ROS) production and calcium mobilization without cytotoxicity. Western blot analysis revealed that A40 down-regulated PMACI-induced MAPK (ERK 1/2, p-38, and JNK) phosphorylation, and the NF-κB translocation from the cytosol to membrane. Moreover, A40 inhibited PMACI-induced interleukin (IL)-1ß, IL-6, and IL-8 production. Anti-allergic activities of A40 were confirmed based on inhibitory effects on IL-4 and tumor necrosis factor alpha (TNF-α) production in compound (com) 48/80-induced rat basophilic leukemia (RBL)-2H3 cells. A40 inhibited ß-hexosaminidase release and cytokine production such as IL-4 and TNF-α produced by com 48/80-stimulated RBL-2H3 cells. Furthermore, it's fraction attenuated the IgE/DNP-induced passive cutaneous anaphylaxis (PCA) reaction in the ears of BALB/c mice. Our results suggest that abalone contains the active fraction, A40 is a potent therapeutic and functional material to treat allergic diseases.

Anti-Inflammatory Food in Asthma Prepared from Combination of Raphanus sativus L., Allium hookeri, Acanthopanax sessiliflorum, and Dendropanax morbiferus Extracts via Bioassay-Guided Selection.

Asthma is a highly prevalent inflammatory disease of the respiratory airways and an increasing health risk worldwide. Hence, finding new strategies to control or attenuate this condition is necessary. This study suggests nutraceuticals that are a combination of herbal plant extracts prepared from Acanthopanax sessiliflorum (AS), Codonopsis lanceolate (CL), Dendropanax morbiferus (DM), Allium hookeri (AH), and Raphanus sativus L. (RS) that can improve immunomodulatory ability through the detoxification and diuresis of air pollutants. Herbal parts (AH whole plant, RS and CL roots, AS and DM stems, and DM leaves) were selected, and four types of mixtures using plant extracts were prepared. Among these mixtures, M2 and M4 exhibited antioxidant activities in potent 2,2-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) and 1,1-diphenyl-ß-picrylhydrazine (DPPH) radical assays. Moreover, M4 exhibited a marked increase in glutathione S-transferase (GST) activity and significantly inhibited the inflammatory mediator, nitric oxide (NO) and proinflammatory cytokines, interleukin (IL)-1ß, IL-6 and tumor necrosis factor (TNF)-α generation. Furthermore, M4 exhibited the strongest antioxidant, hepatoprotective, and anti-inflammatory effects and was selected to prepare the product. Before manufacturing the product, we determined that the active mixture, M4, inhibited gene expression and generation of proinflammatory cytokines IL-1ß, IL-6 and TNF-α in ovalbumin (OVA)-, lipopolysaccharide (LPS)-, and particulate matter (PM)-induced asthmatic rat models. The granular product (GP) was manufactured using M4 along with additives, i.e., lactose, oligosaccharide, stevioside extract, and nutmeg seed essential oils (flavor masking), in a ratio of 1:4 using a granulation machine, dried and ultimately packaged. The GP inhibited the generation of proinflammatory cytokines IL-1ß, IL-6 and TNF-α in OVA-, LPS- and PM-induced asthmatic rat models. These results suggest that GP prepared from a combination of herbal plants (AS, CL, DM, AH and RS) is a potent functional food with anti-inflammatory activity that can be used to treat asthma caused by ambient air pollutants.

A Transcriptomic Response to Lactiplantibacillus plantarum-KCC48 against High-Fat Diet-Induced Fatty Liver Diseases in Mice.

The most prevalent chronic liver disorder in the world is fatty liver disease caused by a high-fat diet. We examined the effects of Lactiplantibacillus plantarum-KCC48 on high-fat diet-induced (HFD) fatty liver disease in mice. We used the transcriptome tool to perform a systematic evaluation of hepatic mRNA transcripts changes in high-fat diet (HFD)-fed animals and high-fat diet with L. plantarum (HFLPD)-fed animals. HFD causes fatty liver diseases in animals, as evidenced by an increase in TG content in liver tissues compared to control animals. Based on transcriptome data, 145 differentially expressed genes (DEGs) were identified in the liver of HFD-fed mice compared to control mice. Moreover, 61 genes were differentially expressed in the liver of mice fed the HFLPD compared to mice fed the HFD. Additionally, 43 common DEGs were identified between HFD and HFLPD. These genes were enriched in metabolic processes, retinol metabolism, the PPAR signaling pathway, fatty acid degradation, arachidonic metabolism, and steroid hormone synthesis. Taking these data into consideration, it can be concluded that L. plantarum-KCC48 treatment significantly regulates the expression of genes involved in hepatosteatosis caused by HFD, which may prevent fatty liver disease.

Novel Bacillus ginsengihumi CMRO6 Inhibits Adipogenesis via p38MAPK/Erk44/42 and Stimulates Glucose Uptake in 3T3-L1 Pre-Adipocytes through Akt/AS160 Signaling.

The health benefits of probiotics have been known for decades, but there has only been limited use of probiotics in the treatment of obesity. In this study, we describe, for the first time, the role of cell-free metabolites (CM) from Bacillus ginsengihumi-RO6 (CMRO6) in adipogenesis and lipogenesis in 3T3-L1 pre-adipocytes. The experimental results show that CMRO6 treatment effectively reduced lipid droplet accumulation and the expression of CCAAT/enhancer-binding protein α and ß (C/EBPα and C/EBPß), peroxisome proliferator-activated receptor γ (PPAR-γ), serum regulatory binding protein 1c (SREBP-1c), fatty acid-binding protein 4 (FABP4), fatty acid synthase (FAS), acetyl CoA carboxylase (ACC), phosphorylated p38MAPK, and Erk44/42. Additionally, CMRO6 treatment significantly increased glucose uptake and phosphorylated Akt (S473), AS160, and TBC1D1 protein expressions. Considering the results of this study, B. ginsengihumi may be a novel probiotic used for the treatment of obesity and its associated metabolic disorders.

Direct observation of kink evolution due to Hund's coupling on approach to metal-insulator transition in NiS2-xSex.

Understanding characteristic energy scales is a fundamentally important issue in the study of strongly correlated systems. In multiband systems, an energy scale is affected not only by the effective Coulomb interaction but also by the Hund's coupling. Direct observation of such energy scale has been elusive so far in spite of extensive studies. Here, we report the observation of a kink structure in the low energy dispersion of NiS2-xSex and its characteristic evolution with x, by using angle resolved photoemission spectroscopy. Dynamical mean field theory calculation combined with density functional theory confirms that this kink originates from Hund's coupling. We find that the abrupt deviation from the Fermi liquid behavior in the electron self-energy results in the kink feature at low energy scale and that the kink is directly related to the coherence-incoherence crossover temperature scale. Our results mark the direct observation of the evolution of the characteristic temperature scale via kink features in the spectral function, which is the hallmark of Hund's physics in the multiorbital system.

Microencapsulation of endophytic LAB (KCC-41) and its probiotic and fermentative potential for cabbage kimchi

The aim of the present study was to isolate novel lactic acid bacteria (LAB) from hairy vetch forage crop and characterize their probiotic and fermentative potential for preparing Korean cabbage kimchi. First, functional characterization of isolated strains such as antagonistic property, auto-aggregation, antibiotic susceptibility, and extracellular enzyme production was performed. The isolated Lactobacillus plantarum KCC-41 strain was able to inhibit pathogenic fungal spore formation. It showed susceptibility to common commercial antibiotics drugs. The selected LAB strain was then subjected to microencapsulation with alginate biopolymer. Its ability to survive in in vitro simulated gastro-intestinal fluid was evaluated. It was also used in the fermentation of cabbage kimchi samples. The encapsulated KCC-41 strain could effectively lead to kimchi fermentation in terms of reducing its pH and dominating bacterial count. It also significantly increased organic acid production than non-encapsulated LAB (KCC-41) for cabbage kimchi samples

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Microencapsulation of endophytic LAB (KCC-41) and its probiotic and fermentative potential for cabbage kimchi.

The aim of the present study was to isolate novel lactic acid bacteria (LAB) from hairy vetch forage crop and characterize their probiotic and fermentative potential for preparing Korean cabbage kimchi. First, functional characterization of isolated strains such as antagonistic property, auto-aggregation, antibiotic susceptibility, and extracellular enzyme production was performed. The isolated Lactobacillus plantarum KCC-41 strain was able to inhibit pathogenic fungal spore formation. It showed susceptibility to common commercial antibiotics drugs. The selected LAB strain was then subjected to microencapsulation with alginate biopolymer. Its ability to survive in in vitro simulated gastro-intestinal fluid was evaluated. It was also used in the fermentation of cabbage kimchi samples. The encapsulated KCC-41 strain could effectively lead to kimchi fermentation in terms of reducing its pH and dominating bacterial count. It also significantly increased organic acid production than non-encapsulated LAB (KCC-41) for cabbage kimchi samples.

Evaluation of probiotic Lactobacillus plantarum against foodborne pathogens and its fermentation potential in improving Lolium multiflorum silage quality.

The objective of this study was to isolate the lactic acid bacteria from fermented silage sample and analyze their antibacterial activities, probiotic properties, and fermentation potential in silage. Eleven lactic acid bacteria (LAB) were selected based on distinct morphologies and preliminary studies. Cell-free supernatant (CFS) was then prepared from the selected strains for antibacterial analysis. L-30 strain and its CFS showed highest inhibition (> 10 mm) against tested foodborne pathogens as compared to other strains. Hereafter, the strain L-30 was named as KCC-30 and used for further studies. KCC-30 can survive in the harsh conditions of GIT such as low pH ( 2) and bile salt environment (oxgal) than standard L. plantarum KACC-91016 (pH 2: 27.2% vs 20.5%; oxgal: 72.3% vs 57.7%, both p < 0.05). In addition, KCC-30 exhibited strong auto-aggregation (68.3% vs 51.5%) and co-aggregation (33% vs 23.9%) properties. For silage experiment, KCC-30 treatment did not alter the nutrient profiles of silage. At the same time, KCC-30 treatment increased the lactic acid content of silage as compared to untreated silage (5.55 DM% vs 3.11 DM%). An increase of lactic acid content in the silage is due to higher lactic acid bacteria population in KCC-30 treated silage (15.33 × 107 CFU/g vs 7.66 × 107 CFU/g) than untreated silage (p < 0.05). Overall data suggested that KCC-30 exhibited strong probiotic potential and improved the quality of Lolium multiflorum silage by increasing the lactic acid level. Therefore, KCC-30 could be considered as potential strain to improve the fermentation quality of L. multiflorum silage.

Forage Crop Lolium multiflorum Assisted Synthesis of AgNPs and Their Bioactivities Against Poultry Pathogenic Bacteria in In Vitro.

Italian ryegrass is one of main feed for livestock animals/birds. It has potential antioxidant metabolites that can improve their health and protect them against various infectious diseases. In this work, we studied synthesis of silver nanoparticles assisted by forage crop Lolium multiflorum as a green synthesis way. Potential antibacterial efficacy of these synthesized nanosized silver nanoparticles against poultry pathogenic bacteria was then studied. Aqueous extract of IRG was used as reducing agent for bio-reduction of silver salt to convert Ag+ to Ag0 metallic nano-silver. Size, shape, metallic composition, functional group, and crystalline nature of these synthesized silver nanoparticles were then characterized using UV-Vis spectrophotometer, FESEM, EDX, FT-IT, and XRD, respectively. In addition, antibacterial effects of these synthesized AgNPs against poultry pathogenic bacteria were evaluated by agar well diffusion method. UV-Vis spectra showed strong absorption peak of 440-450 nm with differ reaction time ranging from 30 min to 24 h. FESEM measurements revealed particles sizes of around 20-100 nm, majority of which were spherical in shape while a few were irregular. These biosynthesized silver nanoparticles using IRG extract exhibited strong antibacterial activities against poultry pathogenic microorganisms, including Pseudomonas aeruginosa, Salmonella typhi, Escherichia coli, and Bacillus subtilis. Overall results confirmed that IRG plant extract possessed potential bioactive compounds for converting silver ions into nanosized silver at room temperature without needing any external chemical for redox reaction. In addition, such synthesized AgNPs showed strong antibacterial activities against pathogenic bacteria responsible for infectious diseases in poultry.

Potential Sustainable Properties of Microencapsulated Endophytic Lactic Acid Bacteria (KCC-42) in In-Vitro Simulated Gastrointestinal Juices and Their Fermentation Quality of Radish Kimchi.

The objective of this study was to investigate alginate microencapsulated lactic acid bacteria (LAB) fermentation quality of radish kimchi sample and its potential survivability in different acidic and alkaline environments. Initially, we isolated 45 LAB strains. One of them showed fast growth pattern with potential probiotic and antifungal activities against Aspergillus flavus with a zone of inhibition calculated with 10, 8, 4mm for the 4th, 5th, and 6th day, respectively. Therefore, this strain (KCC-42) was chosen for microencapsulation with alginate biopolymer. It showed potential survivability in in-vitro simulated gastrointestinal fluid and radish kimchi fermentation medium. The survival rate of this free and encapsulated LAB KCC-42 was 6.85 × 105 and 7.48× 105 CFU/ml, respectively; the viability count was significantly higher than nonencapsulated LAB in simulated gastrointestinal juices (acid, bile, and pancreatin) and under radish kimchi fermentation environment. Kimchi sample added with this encapsulated LAB showed increased production of organic acids compared to nonencapsulated LAB sample. Also, the organic acids such as lactic acid, acetic acid, propionic acid, and succinic acid production in fermented kimchi were measured 59mM, 26mM, 14mM, and 0.6mM of g/DW, respectively. The production of metabolites such as lactic acid, acetic acid, and succinic acid and the bacteria population was high in microencapsulated LAB samples compared with free bacteria added kimchi sample. Results of this study indicate that microencapsulated LAB KCC-42 might be a useful strategy to develop products for food and healthcare industries.

Quantification of major phenolic and flavonoid markers in forage crop Lolium multiflorum using HPLC-DAD

ABSTRACT The objective of this study was to perform preliminary screening of phytochemical compounds and quantification of major phenolics and flavonoid markers in Italian ryegrass extract using HPLC-DAD. Previously, LC-MS analysis has identified different phenolic acids, including caffeic acid, ferulic acid, p-coumaric acid, chlorogenic acid, dihydroxy benzoic acid, propyl gallate, catechin, and six flavonoids including rutin hydroxide, luteolin, kaemferol, vitexin, narcissoside, and myricetin from Italian ryegrass extract. In the present study, Italian ryegrass silage powder was extracted with ethanol: water for 20 min at 90 °C. The extract targeted optimum yield of phenolic acids and flavonoids. Crude phenolic acid and flavonoids were then purified by solid phase extraction method. Purified fractions were then injected into HPLC with a diode-array detector. Quantified concentrations of isolated phenolic acids and flavonoids ranged from 125 to 220 µg/g dry weight. Limits of detection and limits of quantification for all standards (unknown compounds) ranged from 0.38 to 1.71 and 0.48 to 5.19 µg/g dry weight, respectively. Obtained values were compared with previous literatures, indicating that our HPLC-DAD quantification method showed more sensitivity. This method showed better speed, accuracy, and effectiveness compared to previous reports. Furthermore, this study could be very useful for developing phenolic acids and flavonoids from compositions in Italian ryegrass silage feed for pharmaceutical applications and ruminant animals in livestock industries.

Anti-neuroinflammatory effects of tryptanthrin from Polygonum tinctorium Lour. in lipopolysaccharide-stimulated BV2 microglial cells.

This study was conducted to isolate the anti-neuroinflammatory component(s) in the 80% EtOH extract of P. tinctoria, and to investigate underlying molecular mechanism of the anti-neuroinflammatory component(s) in LPS-induced BV2 microglial cells. To isolate the active component(s) in the extract, various chromatographic methods were employed, and the structures of the isolated secondary metabolites were determined mainly by analysis of spectroscopic data such as NMR and MS data. Tryptanthrin (1), isolated from P. tinctoria extract, significantly inhibited the protein expression of iNOS and COX-2, and reduced the levels of their products (NO and PGE2) in LPS-stimulated BV2 microglial cells. Tryptanthrin (1) also downregulated the production of pro-inflammatory cytokines such as TNF-α, IL-6, and IL-1ß. These anti-neuroinflammatory effects of tryptanthrin (1) was elucidated to be correlated with inactivating NF-κB pathway by interrupting the phosphorylation and degradation of the inhibitor of κB-α protein, and inhibiting the DNA binding activity of NF-κB. In addition, tryptanthrin (1) suppressed the activation of p38 MAPK pathway. Furthermore, tryptanthrin (1) inhibited the TLR4 and MyD88 protein expression in LPS-stimulated BV2 microglial cells. Taken together, it was suggested that tryptanthrin (1) have anti-neuroinflammatory effect by regulating TLR4-MyD88-mediated several inflammatory pathways including p38 and NF-κB pathways in LPS-induced BV2 microglial cells.

Roles of Carbohydrate-Binding Module (CBM) of an Endo-ß-1,4-Glucanase (Cel5L) from Bacillus sp. KD1014 in Thermostability and Small-Substrate Hydrolyzing Activity.

An endo-ß-1,4-glucanase gene, cel5L, was cloned using the shot-gun method from Bacillus sp.. The gene, which contained a predicted signal peptide, encoded a protein of 496 amino acid residues, and the molecular mass of the mature Cel5L was estimated to be 51.8 kDa. Cel5L contained a catalytic domain of glycoside hydrolase (GH) family 5 and a carbohydrate-binding module family 3 (CBM_3). Chromatography using HiTrap Q and CHT-II resulted in the isolation of two truncated forms corresponding to 50 (Cel5L-p50) and 35 kDa (Cel5L-p35, CBM_3-deleted form). Both enzymes were optimally active at pH 4.5 and 55°C, but had different half-lives of 4.0 and 22.8 min, respectively, at 70°C. The relative activities of Cel5L-p50 and Cel5L-p35 for barley ß-glucan were 377.0 and 246.7%, respectively, compared to those for carboxymethyl-cellulose. The affinity and hydrolysis rate of pNPC by Cel5L-p35 were 1.7 and 3.3 times higher, respectively, than those by Cel5L-p50. Additions of each to a commercial enzyme set increased saccharification of pretreated rice straw powder by 17.5 and 21.0%, respectively. These results suggest CBM_3 is significantly contributing to thermostability, and to affinity and substrate specificity for small substrates, and that these two enzymes could be used as additives to enhance enzymatic saccharification.

Element-Specific Orbital Character in a Nearly-Free-Electron Superconductor Ag5Pb2O6 Revealed by Core-Level Photoemission.

Ag5Pb2O6 has attracted attentions due to its novel nearly-free-electron superconductivity, but its electronic structure and orbital character of the Cooper-pair electrons remain controversial. Here, we present a method utilizing core-level photoemission to show that Pb 6s electrons dominate near the Fermi level. We observe a strongly asymmetric Pb 4 f 7/2 core-level spectrum, while a Ag 3d 5/2 spectrum is well explained by two symmetric peaks. The asymmetry in the Pb 4 f 7/2 spectrum originates from the local attractive interaction between conducting Pb 6s electrons and a Pb 4 f 7/2 core hole, which implies a dominant Pb 6s contribution to the metallic conduction. In addition, the observed Pb 4 f 7/2 spectrum is not explained by the well-known Doniach-Sunjic lineshape for a simple metal. The spectrum is successfully generated by employing a Pb 6s partial density of states from local density approximation calculations, thus confirming the Pb 6s dominant character and free-electron-like density of states of Ag5Pb2O6.

Vaccinium angustifolium Root Extract Suppresses FcɛRI Expression in Human Basophilic KU812F Cells.

Vaccinium angustifolium, commonly known as the lowbush blueberry, is a rich source of flavonoids, with which various human physiological activities have been associated. The present study focuses on the investigation of the effect of the methanolic extract of V. angustifolium root extract (VAE) on high affinity immunoglobulin E receptor (FcɛRI) α chain antibody (CRA-1)-induced allergic reaction in human basophilic KU812F cells. The total phenolic content of VAE was found to be 170±1.9 mg gallic acid equivalents/g. Flow cytometry analysis revealed that the cell surface expression of FcɛRI was suppressed in a concentration-dependent manner upon culture with VAE. Reverse-transcriptase polymerase chain reaction analysis showed that the mRNA level of the FcɛRI α chain was reduced in a concentration-dependent manner as a result of VAE treatment. Western blot analysis revealed that the protein expression of FcɛRI and the phosphorylation of extracellular signal-regulated kinases (ERK) 1/2 were concentration-dependently inhibited by VAE. We determined that VAE inhibited anti-CRA-1-induced histamine release, in addition to the elevation of intracellular calcium concentration ([Ca2+]i), in a concentration-dependent manner. These results indicate that VAE may exert an anti-allergic effect via the inhibition of calcium influx and histamine release, which occurs as a result of the down-regulation of FcɛRI expression through inhibition of ERK 1/2 activation.

Relationship between ion migration and interfacial degradation of CH3NH3PbI3 perovskite solar cells under thermal conditions.

Organic-inorganic hybrid perovskite solar cells (PSCs) have been extensively studied because of their outstanding performance: a power conversion efficiency exceeding 22% has been achieved. The most commonly used PSCs consist of CH3NH3PbI3 (MAPbI3) with a hole-selective contact, such as 2,2',7,7'-tetrakis(N,N-di-p-methoxyphenylamine)-9,9-spiro-bifluorene (spiro-OMeTAD), for collecting holes. From the perspective of long-term operation of solar cells, the cell performance and constituent layers (MAPbI3, spiro-OMeTAD, etc.) may be influenced by external conditions like temperature, light, etc. Herein, we report the effects of temperature on spiro-OMeTAD and the interface between MAPbI3 and spiro-OMeTAD in a solar cell. It was confirmed that, at high temperatures (85 °C), I- and CH3NH3+ (MA+) diffused into the spiro-OMeTAD layer in the form of CH3NH3I (MAI). The diffused I- ions prevented oxidation of spiro-OMeTAD, thereby degrading the electrical properties of spiro-OMeTAD. Since ion diffusion can occur during outdoor operation, the structural design of PSCs must be considered to achieve long-term stability.

Electronic Structure of the Kitaev Material α-RuCl3 Probed by Photoemission and Inverse Photoemission Spectroscopies.

Recently, α-RuCl3 has attracted much attention as a possible material to realize the honeycomb Kitaev model of a quantum-spin-liquid state. Although the magnetic properties of α-RuCl3 have been extensively studied, its electronic structure, which is strongly related to its Kitaev physics, is poorly understood. Here, the electronic structure of α-RuCl3 was investigated by photoemission (PE) and inverse-photoemission (IPE) spectroscopies. The band gap was directly measured from the PE and IPE spectra and was found to be 1.9 eV, much larger than previously estimated values. Local density approximation (LDA) calculations showed that the on-site Coulomb interaction U could open the band gap without spin-orbit coupling (SOC). However, the SOC should also be incorporated to reproduce the proper gap size, indicating that the interplay between U and SOC plays an essential role. Several features of the PE and IPE spectra could not be explained by the results of LDA calculations. To explain such discrepancies, we performed configuration-interaction calculations for a RuCl63- cluster. The experimental data and calculations demonstrated that the 4d compound α-RuCl3 is a Jeff = 1/2 Mott insulator rather than a quasimolecular-orbital insulator. Our study also provides important physical parameters required for verifying the proposed Kitaev physics in α-RuCl3.

Potential Application of p-Coumaric Acid on Differentiation of C2C12 Skeletal Muscle and 3T3-L1 Preadipocytes-An in Vitro and in Silico Approach.

Coumaric acid (CA) is a phenolic acid of the hydroxycinnamic acid family, and it has many biological functions such as anti-oxidant, anti-inflammatory, antidiabetic, anti-ulcer, anti-platelet, anti-cancer activities, etc. In the present study, we planned to analyse the potential molecular function of CA on skeletal muscle and preadipocytes differentiation using PCR and Western blot techniques. First, we analysed the impact of CA on C2C12 skeletal muscle differentiation. It revealed that CA treatment inhibited horse serum-induced skeletal muscle differentiation as evidenced by the decreased expression of early myogenic differentiation markers such as Myogenin and myoD via the AMP activated protein kinase- alpha AMPK-α mediated pathway. Furthermore, the level of lipid accumulation and changes in genes and protein expressions that are associated with lipogenesis and lipolysis were analyzed in 3T3-L1 cells. The Oil Red O staining evidenced that CA treatment inhibited lipid accumulation at the concentration of 0.1 and 0.2 mM. Furthermore, coumaric acid treatment decreased the expression of main transcriptional factors such as CCAAT/enhancer binding protein-alpha (C/EBP-α) and peroxisome proliferator-activated receptor gamma-2 (PPAR-γ2). Subsequently, CA treatment decreased the expression of sterol regulatory element binding protein-1 (SREBP-1), fatty acid synthase (FAS), acetyl CoA carboxylase (ACC) and adiponectin. Finally, we identified conformational changes induced by CA in PPAR-γ2 using computational biology tools. It revealed that CA might downregulate the PPAR-γ2 expression by directly binding with amino acids of PPAR-γ2 by hydrogen at 3.26 distance and hydrophobic interactions at 3.90 contact distances. These data indicated that CA suppressed skeletal muscle and preadipocytes differentiation through downregulation of the main transcriptional factors and their downstream targets.

Antitumor Effect of the Mannich Base(1,3-bis-((3-Hydroxynaphthalen-2-yl)phenylmethyl)urea) on Hepatocellular Carcinoma.

The present study was designed to evaluate the antitumor effects of the synthetic Mannich base 1,3-bis-((3-hydroxynaphthalen-2-yl)phenylmethyl)urea (1,3-BPMU) against HEP-G2 hepatoma cells and diethylnitrosamine (DEN)-induced hepatocarcinoma (HCC) in albino rats. In vitro analysis results revealed that 1,3-BPMU showed significant cytotoxicity and cell growth inhibition in HEP-G2 hepatoma cells in a concentration-dependent manner. Furthermore, flow cytometry results indicated that 1,3-BPMU enhanced early and late apoptosis. The maximum apoptosis was exhibited at a concentration of 100 µg/mL of 1,3-BPMU. In in vivo analysis, DEN treatment increased the content of nucleic acids, LPO and the activities of AST, ALT, ALP, LDH, γGT and 5'NT with decreased antioxidant activity as compared to control rats. However, 1,3-BPMU treatment to DEN-induced rats decreased the content of nucleic acids, LPO and the activities of AST, ALT, ALP, LDH, γGT and 5'NT and increased the activities of SOD, CAT, GPx, GST and GR (p < 0.05). Furthermore, 1,3-BPMU enhanced the apoptosis via upregulation of caspase-3 and caspase-9 and the downregulation of Bcl-2 and Bcl-XL mRNA expression as compared to DEN-induced rats. Histological and ultrastructural investigation showed that 1,3-BPMU treatment renovated the internal architecture of the liver in DEN-induced rats. In this study, the molecular and pre-clinical results obtained by treatment of DEN-induced rats with 1,3-BPMU suggested that 1,3-BPMU might be considered as an antitumor compound in the future.

Achyranthes japonica Nakai Water Extract Suppresses Binding of IgE Antibody to Cell Surface FcɛRI.

Achyranthes japonica Nakai (AJN) water extract has a variety of physiological properties, including anti-diabetic, anti-cancer, anti-inflammatory, anti-microbial, and anti-oxidative activities. In the present study, the inhibitory effects of AJN extract were investigated in high affinity immunoglobulin E receptor (FcɛRI)-mediated KU812F cells activation. AJN extract showed suppressive effects on histamine release and intracellular calcium [Ca2+]i elevation from anti-FcɛRI antibody (CRA-1)-stimulated cells in a dose-dependent manner. Flow cytometric analysis showed that AJN extract treatment caused a dose-dependent decrease in the cell surface FcɛRI expression and the binding between the cell surface FcɛRI and the IgE antibody. Moreover, reverse transcription-polymerase chain reaction analysis showed that levels of the mRNA for the FcɛRI α chain was decreased by treatment with AJN extract. These results indicate that AJN extract may exert anti-allergic effects via the inhibition of calcium influx and histamine release, which occurs as a result from the down-regulation of the binding of IgE antibody to cell surface FcɛRI. This mechanism may occur through FcɛRI expression inhibition.