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The purpose of this research was to identify metabolite change during barley (Hordeum vulgare) germination and reveal active principles for the anti-wrinkle activity. Barley was germinated with deionized water (DW) and mineral-rich water (MRW) for the comparison of the effect of mineral contents on the metabolites changes during germination. The effects of germinated barley extracts (GBEs) on collagen production and collagenase inhibition were evaluated in vitro using human dermal fibroblasts (HDFs). A pronounced anti-wrinkle activity was observed in the test group treated with the MRW-GBEs. In order to find out the active components related to the anti-wrinkle activity, an orthogonal projection to latent structure-discriminant analysis (OPLS-DA) was performed, using the data from secondary metabolites profiling conducted by UPLC–PDA–ESI–MS. The anti-wrinkle activity of MRW-GBEs was revealed to be associated with the increase of oligomeric compounds of procyanidin and prodelphinidin, indicating that it can be used as an active ingredient for anti-wrinkle agents.

Objective: To investigate the anti-senescence effect of 3-bromo-4,5- dihydroxybenzaldehyde (BDB) from Polysiphonia morrowii Harvey in human dermal fibroblasts (HDF). Methods: HDF were subjected to treatment of BDB and then treated with hydrogen peroxide (H2O2) to induce premature senescence. Senescence-associated β-galactosidase (SA-β-gal) activity in HDF was determined using the SA-β-gal staining method. Intracellular reactive oxygen species (ROS) production was measured using the 2’,7’-dichlorodihydrofluorescein diacetate assay. Western blotting assay was performed to assess the level of antioxidant enzyme glutathione peroxidase 1 (GPX1). In addition, intracellular collagen and collagenase contents were analyzed using the respective ELISA kits. Elastase activity in HDF supernatants was measured from p-nitroaniline release and normalized using total protein content. Results: Treatment of HDF with H2O2 increased the activity of SAP-gal, but BDB pre-treatment resulted in the reduction of SA-β- gal activity. Moreover, BDB significantly reduced H2O2-induced intracellular ROS production. BDB also markedly increased the level of GPX1, which was inhibited by 400 µM of H2O2. Furthermore, in in vitro study, BDB significantly increased intracellular collagen content and decreased matrix metalloproteinase-1 and elastase activities in HDF. Conclusions: Our results demonstrate that BDB shows anti- senescence and anti-wrinkle activities in vitro.

We investigated the protective effects of 3-bromo-4,5-dihydroxybenzaldehyde (BDB) from Polysiphonia morrowii Harvey against hydrogen peroxide (H2O2)-induced apoptosis in Vero cells. BDB exhibited scavenging activity for DPPH, hydroxyl, and alkyl radicals. BDB also inhibited H2O2-induced lipid peroxidation, cell death, and apoptosis in Vero cells by inhibiting the production of ROS. To evaluate the molecular mechanisms of apoptosis inhibition, the expression of Bax/Bcl-xL and NF-κB was assessed by western blot assay. BDB significantly suppressed the cleavage of caspase-9 and PARP and reduced Bax levels in H2O2-induced Vero cells. Besides, BDB suppressed the phosphorylation of NF-κB and the translocation of p65 in H2O2-induced cells. Furthermore, we evaluated the effect of BDB on ROS production, cell death, and lipid peroxidation in an H2O2-stimulated zebrafish embryo model. Taken together, these results indicated that ROS generation and cell death were significantly inhibited by BDB in zebrafish embryos, thereby proving that BDB exerts excellent antioxidant activity in vitro and in vivo.

The anti-inflammatory effects of 3‑bromo‑5‑(ethoxymethyl)‑1,2‑benzenediol (BEMB) from Polysiphonia morrowii were evaluated in lipopolysaccharide (LPS)-induced RAW264.7 cells and zebrafish embryo. BEMB showed anti-inflammatory effects by inhibiting the production of nitric oxide (NO) and reactive oxygen species (ROS), and the expression of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) in the LPS-activated RAW264.7 cells and zebrafish embryo without cytotoxicity. Moreover, BEMB suppressed the protein and mRNA expression levels of nuclear factor (NF)-κB (p65 and inhibitor of NF-κB [IκB]-A) in RAW264.7 cells and zebrafish embryo, respectively. Collectively, the results of this study indicate that BEMB suppressed the production of pro-inflammatory mediators such as NO, iNOS, and COX-2 as well as their regulation in LPS-induced RAW264.7 cells and zebrafish embryos by inhibiting ROS production and NF-κB expression. Therefore, this study suggests that BEMB could be a potential anti-inflammatory agent for the treatment of inflammatory diseases.

In this study, trans-1,3-diphenyl-2,3-epoxypropane-1-one (DPEP) was evaluated for its anti-inflammatory activity in lipopolysaccharide (LPS)-stimulated zebrafish embryos. In the present study, DPEP exhibited potential protective effect in the zebrafish embryos as confirmed by survival rate. DPEP acts as an effective agent against reactive oxygen species (ROS) formation induced by LPS. Moreover, DPEP effectively inhibited LPS-induced nitric oxide (NO) production in zebrafish embryos. In addition, DPEP significantly reduced the expression of inducible NOS (iNOS) and cycloxygenase 2 (COX-2), which generate NO as a key mediator of inflammation in a concentration-dependent manner. According to these results, DPEP could be considered an effective anti-inflammatory agent, which might be further developed as a functional ingredient. This is the first report of the anti-inflammatory activity of DPEP in the LPS-stimulated zebrafish model.

Tumor-associated macrophages (TAMs) are major components of tumor microenvironment that promote invasion and metastasis of cancer cells. In this study, we investigated the effect of TAMs on phenotypic conversion of non-neoplastic MCF10A human breast epithelial cells using an indirect co-culture system. Co-culture with TAMs induced epithelial-to-mesenchymal transition, invasive phenotype, and MMP-9 upregulation in MCF10A cells. Comparative proteomic analysis revealed that endoplasmic reticulum oxidoreductase (ERO)1-α was increased in MCF10A cells co-cultured with TAMs compared to that in mono-cultured cells. ERO1-α was crucial for TAMs-induced invasive phenotype and MMP-9 upregulation involving transcription factors c-fos and c-Jun. Cytokine array analysis showed that levels of interleukin (IL)-6, C-X-C motif ligand (CXCL)1, C-C motif ligand (CCL)2, growth-regulated protein (GRO), IL-8, and granulocyte-macrophage colony-stimulating factor (GM-CSF) were increased in conditioned media of co-cultured cells. Among these cytokines increased in conditioned media of co-cultured cells, CCL2 was secreted from TAMs, leading to induction of ERO1-α, MMP-9 upregulation, and invasiveness in MCF10A cells. Our findings elucidated a molecular mechanism underlying the aggressive phenotypic change of non-neoplastic breast cells by co-culture with TAMs, providing useful information for prevention or treatment of recurrent breast cancer.

In this study, the roles of reactive oxygen species (ROS) and NF-κB on inflammation induction in lipopolysaccharide (LPS)-stimulated zebrafish embryos were evaluated using N-acetyl-l-cysteine (NAC) and pyrrolidine dithiocarbamate (PDTC), specific inhibitors of ROS and NF-κB, respectively. LPS-stimulated zebrafish embryos showed increasing production of NO and ROS and expression of iNOS and COX-2 protein, compared to a control group without LPS. However, NAC significantly inhibited production of NO and ROS and markedly suppressed expression of iNOS and COX-2 protein in LPS-stimulated zebrafish embryos. The mRNA expressions of NF-κB such as p65NF-κB and IκB-A were significantly increased after LPS stimulation, whereas PDTC attenuated mRNA expression of NF-κB. PDTC also inhibited production of NO and reduced expression of iNOS and COX-2 protein in LPS-stimulated zebrafish embryos. Taken together, these results indicated that LPS increases pro-inflammatory mediators in zebrafish embryos through ROS and NF-κB regulation.

An oscillating multi-mold simulator with embedded thermocouples was used to study the initial solidification of medium carbon steels and crystallization characteristics of the mold flux. Casting speed variations in the simulator from 0.7 m/min to 1.4 m/min at fixed oscillation frequency and stroke resulted in higher copper mold temperatures. Frequency modifications from 2.5 Hz to 5.0 Hz and stroke changes from 8.1 mm to 5.4 mm at fixed casting speeds also resulted in higher copper mold temperatures. Surface profile analysis of as-cast steel strips showed characteristic oscillation marks comparable to the narrow faces of the industrial cast slabs. The apparent effect of casting variables on the temperature and surface profiles during the solidification of the medium carbon steels could be correlated to the variations in the negative strip time and subsequent changes in the extent of mold flux infiltration. Back scattered scanning electron microscope analysis of the full length of the retrieved flux film after casting showed cuspidine crystallization ratio that increased from the upper to lower portion of the flux film. This dynamic crystallization and growth of the cuspidine phase increases as the flux is sustained at high temperatures for longer periods. Additional experiments with industrial fluxes designed for soft cooling of medium carbon steel grades showed comparable infiltration thickness of the flux, but the crystallization characteristics were significantly different, which could have a significant impact on the heat transfer rate and mechanism through the flux film.

An inverted water-cooled multi-mold continuous casting simulator was used to investigate initial solidification of low-carbon steels and crystallization of mold flux. Embedded mold thermocouples showed characteristic temperature profiles dependent on parameters including casting speed, oscillation frequency, and stroke. Higher maximum temperatures for thermocouples at higher casting speeds, higher frequencies, and lower stroke lengths were observed. The surface of the as-cast steel strips showed oscillation marks similar to those of industrially cast slabs and higher casting speeds resulted in shallower oscillation marks. The measured pitch agreed well with the theoretical pitch suggesting the multi-mold simulator to be a cost-effective alternative to pursue fundamental studies on initial solidification in the mold. Analysis of the mold flux taken between the copper mold and solidified steel shell showed highly dendritic uni-directional crystallization occurring within the flux film suggesting that the heat transfer direction is dominantly horizontal towards the water-cooled copper mold. In addition, the solidified flux located at the upper to lower part of the mold suggested morphological differences in the size and shape of the crystalline phases indicating that crystallization ratio can increase depending upon the retention in the mold and subsequently decrease radiative heat transfer as the flux traverses down the mold.

In the continuous casting process, molten flux film locates between mold and solidified shell. The film has two important roles. One is lubrication of two solids. The other role is controlling heat transfer from the shell to the mold. The shape of the liquid flux film is one of the most important factors for the film to execute both roles properly. However, the number of the studies about the shape of the film has been rare and results are quite different from each other. In the present study, a new approach is applied to determine the shape of the film. This approach is based on the force valence between static pressure of molten steel and internal pressure of liquid flux film and solving heat and momentum equations in the flux film. The results of the present study well explained the experimental results of laboratory and field operations.