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β-glucans, the class of biological response modifier has unceasing attention, not only for its immune stimulating but also for its role as prebiotics, modulator of physiological events etc. and is widely used in the treatment of cancer, diabetes, gastrointestinal disorders, cardiovascular diseases etc. However, β-glucan with different physiochemical properties is found to have discrete clinical functions and thus careful selection of the types of β-glucan plays pivotal role in providing significant and expected clinical outcome. Herein this review, we presented the factors responsible for diverse functional properties of β-glucan, their distinct mode of actions in regulating human health etc. Further, clinical aspects of different β-glucans toward the management of wound care, metabolic dysbiosis, fatty liver disorders and endurance training associated energy metabolism were compiled and exhibited in detail.

Ambient air pollution, in particular, particulate matter (PM) pollution imposes serious health concerns such as hospitalization and premature deaths, worldwide. While commercial breathing masks are in use for protection against this hazardous issue, yet their efficiency in filtering PM was not up to the par, besides several other discomforts such as poor breathability due to reduced air flow, sweat production etc. In this study, commercial face mask coated with β-glucan, a high molecular weight polymer is tested for its efficacy in filtering PM. Quantification of PM before and after filtration and microscopic observation (using scanning electron microscopy (SEM)) of the fabric used in filtering the dust pollutants (generated from wood chips and cigarette) showed that β-glucan coated fabric were significantly efficient in capturing PM (size of 10 and 2.5 μm in diameter) than that of the untreated control fabric, wherein the former had filtration efficacy with fold increase of 11.6 and 2.6 towards capturing PM2.5 and PM10 respectively than the latter. Thus, β- glucan coated fabric was found to be effective in filtering PM.

Myricetin is a commonly found dietary flavonoid. In the present study, we investigated the effects of myricetin on migration and invasion of radioresistant lung cancer cells (A549‐IR). Transcriptome analysis of A549‐IR cells identified several differentially expressed genes (DEGs) in A549‐IR cells compared to parental A549 cells. Functional enrichment analysis revealed that most of the DEGs were linked with PI3K‐AKT signaling, proteoglycans, focal adhesion, and ECM–receptor interactions. A549‐IR cells demonstrated enhanced migratory potential with increased expression of vimentin, snail and slug, and reduced expression of E‐cadherin. A549‐IR cells exposed to myricetin displayed reduced migration and suppressed MMP‐2 and MMP‐9 expression. Notably, myricetin inhibited the phosphorylation of focal adhesion kinase (FAK) and altered the F‐actin/G‐actin ratio in A549‐IR cells, without modulation of EMT markers. These findings suggest that myricetin can inhibit migration of A549‐IR cells by suppressing MMP‐2 and MMP‐9 expressions through inhibition of the FAK‐ERK signaling pathway., Transcriptome analysis of radioresistant lung cancer cells (A549‐IR) identified several differentially expressed genes compared to A549 parental cells. Myricetin inhibited invasion and migration of A549‐IR cells by reducing the expression of matrix metalloproteinases (MMP‐2 and MMP‐9) through inhibition of the FAK‐ERK signaling pathway.

It has been proven that β-glucan produced by Schizophyllum commune has beneficial effects on obesity, obesity-associated constipation, and colitis conditions; however, the protective effect of the compound on host at basal state is yet to be investigated. C57BL/6 J mice were fed with a normal diet (ND), normal diet supplemented with 3 g/kg (BG_low), and 5 g/kg (BG_high) of β-glucan (BG) for 12 weeks. Body weight, food and water intake and fecal status were monitored weekly. Intestine was stained by Periodic acid–Schiff (PAS) and Alcian Blue to evaluate the mucin layer thickness and goblet cell population. Morphological changes in internal organs and intestinal motility were also assessed, while serum biomarkers for liver injury and glucose level were analyzed. On the other hand, fecal microbiota and associated metabolic activities were also investigated. β-glucan bulked feces, decreased fecal moisture, and proliferated goblet cells resulted in a thickened lubricating mucin layer; however, the intestinal transit rate did not increase. The abundance of beneficial bacteria was increased while the harmful strains was decreased in a dose-dependent manner by the effect of β-glucan. Specific short chain fatty acid (SCFA)-producing strains, such as Roseburia, Ruminococcus, and Bifidobacteria, were selectively increased by β-glucan. In addition, consumption of β-glucan lowered level of obesity-associated biomarkers. Schizophyllum commune β-glucan showed an insignificant change in transit rate in healthy conditions when compared with obesity, despite similar effects on increasing mucus production and bulked feces. Nonetheless, the outcomes proposed protective effects against obesity, diabetes, inflammatory bowel diseases (IBD), and constipation, in which the modification of the gut microbiota by β-glucan is the largest contributor.

Objective: We propose that sirtuin (SIRT) may induce a pro-apoptotic effect by deacetylating transcription factors in A549 cells: depletion of sirtuin-1 (SIRT1) induced cell cycle arrest in cisplatin-resistant A549 (A549/CADD) cells. Methods: Protein and mRNA levels of SIRT1 were investigated using western blot and RT-PCR. In A549 and A549/CADD cells, the cytotoxicity of cisplatin administration was evaluated by MTT assay, proliferation was measured by ECIS, and the cell cycle distribution was analyzed using FACS. Cells were transfected with pcDNA3.1-Myc-SIRT1 or pcDNA3.1-Myc-Control vectors to analyze the impact of SIRT-1 on cisplatin induced drug resistance. SIRT1 localization was studied using immunofluorescence analysis. In addition, immunoprecipitation and 20S proteasome activity assay were performed to examine the relationship of SIRT1 with the proteasome complex. Results: A549/CADD cells exhibited a mesenchymal-like cell characteristic. SIRT1 expression was markedly decreased in A549/CADD cells. We observed that cisplatin regulates p53 stability through the depletion of ubiquitination following SIRT1 downregulation. Furthermore, cisplatin treatment increased proteasomal activity and significantly decreased cytoplasmic SIRT1 protein levels in A549/CADD cells. Conclusion: In this study, we found SIRT1 to be depleted in A549/CADD cells and also determined the underlying resistance mechanism which may act as novel therapeutic targets in overcoming drug resistance.

With poor prognosis and aberrant lung remodeling, pulmonary fibrosis exhibits worldwide prevalence accompanied by an increase in burden in terms of hospitalization and death. Apart from genetic and non-genetic factors, fibrosis occurs as a side effect of bleomycin antineoplastic activity. Elucidating the cellular and molecular mechanism could help in the development of effective anti-fibrotic treatment strategies. In the present study, we investigated the underlying mechanism behind bleomycin-induced fibrosis using human alveolar epithelial cells (A549 cells). On the basis of the experimental observation, it was demonstrated that with transforming growth factor-β (TGF-β) as a central mediator of fibrosis progression, a cross-talk between epithelial-mesenchymal transition (EMT) and senescence upon bleomycin treatment occurs. This results in the advancement of this serious fibrotic condition. Fibrosis was initiated through integrin activation and imbalance in the redox state (NOX expression) of the cell. It progressed along the TGF-β-mediated non-canonical pathway (via ERK phosphorylation) followed by the upregulation of α-smooth muscle actin and collagen synthesis. Additionally, in this process, the loss of the epithelial marker E-cadherin was observed. Furthermore, the expressions of senescence markers, such as p21 and p53, were upregulated upon bleomycin treatment, thereby intensifying the fibrotic condition. Accordingly, the molecular pathway mediating the bleomycin-induced fibrosis was explored in the current study.

Objective: Recognized as pathogen-associated molecular patterns (PAMPs), β-glucans, a naturally occurring heterogeneous group of polysaccharides, were investigated for their ability to accelerate wound healing in the form of high water-retaining hydrogel dressing. Approach: Full-thickness wounds on the dorsal side of mice created using a 5-mm biopsy punch were treated with β-glucan-based hydrogel for 2 weeks. Standardized photographs of the wound site were taken at regular time intervals to calculate the percentage of wound closure. Tissues isolated from the wound area were subjected to histological examination and immunoblot analysis. Results: β-Glucan-based hydrogel significantly accelerated the duration of wound healing and enhanced the development of skin appendages in the regenerated skin tissue. Increased expression of transforming growth factor-β3 in the skin tissue isolated from the healed wound site indicated that skin regeneration rather than skin repair occurred, thereby minimizing cutaneous scarring. The expression level of cytokeratin 10 and cytokeratin 14 in the isolated skin tissue revealed that the wounds treated with hydrogel showed proper differentiation and proliferation of keratinocytes in the epidermal layer. Innovation: Immunomodulating β-glucan (responsible for fighting infections at the wound site, and enhancing the migration and proliferation of keratinocytes and fibroblasts) in the form of a three-dimensional hydrogel membrane that retains a high water content (responsible for cooling and soothing effect around the wound site, thereby reducing pain) was prepared and analyzed for its effects on the cutaneous wound healing mechanism. Conclusion: β-Glucan-based hydrogels are promising as wet wound dressings in the health care industry.

The effects of enzymatic hydrolysates of Hippocampus abdominalis on skeletal muscle growth in C2C12 myoblasts and zebrafish were investigated. The hydrolysates were prepared by enzymatic extraction...

Probiotics and prebiotics are commonly used to improve the gut microbiota. Since prebiotics can support the growth of probiotics, co-administration of these is called synbiotics. It has been demonstrated that obesity-induced gut dysbiosis can worsen inflammatory bowel disease symptoms. This study evaluated how modulation of gut microbiota with Schizophyllum commune-derived β-glucan (BG), probiotics (PRO), and synbiotics containing both BG and PRO (SYN) could improve the symptoms of obesity-associated colitis and hepatic manifestation. Mice were fed a normal diet (ND), high-fat diet (HFD), and HFD with different additives (BG, PRO, and SYN) for 12 weeks, followed by 5 days of colitis induction. Mice were sacrificed before and after colitis induction. During the experiment, body weight, food and water consumption, and rectal bleeding were monitored. Proteins from the colon were subjected to western blotting, and serum biomarkers such as alanine transaminase, alkaline phosphatase, triglycerides, and total cholesterol were analyzed. Colon and liver samples were sectioned for histological analysis. The fecal microbiota was analyzed based on partial 16S rRNA gene sequences. Although BG and PRO secured intestinal tight junctions, these two treatments did not modulate inflammatory cell infiltration and inflammatory markers (i.e., IL-6 and TNF-α). In contrast, SYN demonstrated stronger and broader effects in reducing colonic inflammation. While BG treatment increased the abundance of indigenous Lactobacillus, PRO treatment decreased bacterial diversity by suppressing the growth of several species of bacteria. SYN treatment groups, however, supported the growth of both indigenous and supplemented bacteria while maintaining bacterial diversity. Obesity-associated colitis can be improved by modulating gut bacteria with β-glucan and probiotics. The co-administration of both outperformed β-glucan and probiotic treatment alone by fostering both indigenous and supplemented probiotic strains.

Synbiotics synergistically favors beneficial effects of prebiotics and probiotics towards host metabolic health by modulating gut ecosystem. In this study, we sought to examine the effects of prebiotics (Schizophyllum commune derived β-(1,3/1,6)-glucan), probiotics (concoction made of eight different bacterial strains) and synbiotics (prebiotics + probiotics) on gut microbiota and its associated metabolic functions through 16S rRNA gene sequences analysis. Results showed that probiotics strains used in this study were detected more in the synbiotic and probiotic treatments, while prebiotic dietary intervention increased the total bacterial abundance and metabolisms related to host immune strengthening. Probiotics and synbiotics dietary interventions enhanced similar metabolisms relating to butanediol and s-adenosyl-l-methionine biosynthesis. Probiotics treatment also showed depleted metabolic activities related to SCFA productions, that were not depleted in prebiotics treatment. With varying differential abundance patterns and metabolic activities across the treatments, our results suggest that synbiotic treatment provide more beneficial effects over probiotics and prebiotics.

Background:Interleukin-6 (IL-6) is involved in fibroblast-like synoviocyte (FLS) activation and promotes pannus formation and bone and cartilage destruction in rheumatoid arthritis (RA). Cysteine-rich 61 (Cyr61) protein regulates cell proliferation, migration, and differentiation. The aim of this study was to investigate the role of Cyr61 in RA-FLS migration and invasion after IL-6 stimulation. Methods:Western blotting and reverse transcription-polymerase chain reaction were used to examine protein and mRNA levels of Cyr61, matrix metalloproteinases (MMPs), and other signalling proteins. Knockdown of gene expression was performed with siRNA, and RNA sequencing was performed for differential gene analysis. Migration and invasion were assessed by wound healing and Boyden chamber assays. Results:Cyr61 levels were elevated in FLSs from RA patients compared to those in osteoarthritis patients. Control and IL-6-treated FLSs showed differential gene expression. IL-6 stimulated protein synthesis of Cyr61, which was attenuated by the extracellular signal-related kinase 1/2 (ERK 1/2) inhibitor, PD98059, and knockdown of early growth response 3 (EGR3), but not of JUN. IL-6-induced Cyr61 protein synthesis increased expression of MMP2. Cyr61 promoted FLS migration and invasion in an autocrine manner. Knockdown of CYR61 and a neutralizing antibody attenuated Cyr61 synthesis and IL-6 induced FLS migration. Conclusions:By modulating the ERK/EGR3 pathway, IL-6 stimulated Cyr61 production and in turn increasedinvasiveness of FLS. Our data suggest that Cyr61 might be a potential target to prevent the progression of joint damage in RA.

Background Interleukin-6 (IL-6) is involved in fibroblast-like synoviocyte (FLS) activation and promotes pannus formation and bone and cartilage destruction in rheumatoid arthritis (RA). Cysteine-rich 61 (Cyr61) protein regulates cell proliferation, migration, and differentiation. The aim of this study was to investigate the role of Cyr61 in RA-FLS migration and invasion after IL-6 stimulation. Methods Western blotting, immunohistochemistry, reverse transcription-polymerase chain reaction, and real time-polymerase chain reaction were used to examine protein and mRNA levels of Cyr61, matrix metalloproteinases (MMPs), and other signalling proteins. Knockdown of gene expression was performed with siRNA, and RNA sequencing was performed for differential gene analysis. Migration and invasion were assessed by wound healing and Boyden chamber assays. Results Cyr61 levels were elevated in FLSs from RA patients compared to those in osteoarthritis patients. Control and IL-6-treated FLSs showed differential gene expression. IL-6 stimulated protein synthesis of Cyr61, which was attenuated by the extracellular signal-related kinase 1/2 (ERK 1/2) inhibitor, PD98059, and knockdown of early growth response 3 (EGR3), but not of JUN. IL-6-induced Cyr61 protein synthesis increased expression of MMP2. Cyr61 promoted FLS migration and invasion in an autocrine manner. Knockdown of CYR61 and a neutralising antibody attenuated Cyr61 synthesis and IL-6-induced FLS migration. Conclusions By modulating the ERK/EGR3 pathway, IL-6 stimulated Cyr61 production and in turn increased invasiveness of FLS. Our data suggest that Cyr61 might be a potential target to prevent the progression of joint damage in RA.

Epithelial-to-mesenchymal transition (EMT) promulgates epithelial cell associated disease-defining characteristics in tumorigenesis and organ fibrosis. Growth factors such as epidermal growth factor and fibroblast growth factor in addition to cytokines such as transforming growth factor-&beta, 1 (TGF-&beta, 1) is said to play a prominent role in remodeling related pathological events of cancer progression such as invasion, metastasis, apoptosis, EMT, etc. through redox related cellular secondary messengers, in particular the reactive oxygen species (ROS). However, the signaling cascade underlying the redox mechanism and thereby the progression of EMT remains largely unknown. In this study, upon TGF-&beta, 1 treatment, we observed an induction in NOX isoforms&mdash, NOX2 and NOX4&mdash, that have time (early and late) and cellular localization (nucleus and autophagosome co-localized) dependent effects in mediating EMT associated cell proliferation and migration through activation of the focal adhesion kinase (FAK)/SRC pathway in HeLa, human cervical cancer cells. Upon silencing NOX2/4 gene expression and using the SRC inhibitor (AZD0530), progression of TGF-&beta, 1 induced EMT related cellular remodeling, extra cellular matrix (ECM) production, cell migration and invasion, got significantly reverted. Together, these results indicate that NOX2 and NOX4 play important, albeit distinct, roles in the activation of cytokine mediated EMT and its associated processes via tyrosine phosphorylation of the FAK/SRC pathway.

Treatment towards wound healing, a complex and dynamic process, has been given a great deal of efforts in the last few decades. Focus has been imposed on developing wound dressings that meet the requirements for proper wound healing. In this study, hydrogel made from blends of poly (vinyl alcohol) and β-1,6-branched-β-1,3-glucan (beta-glucan) were synthesized by modified solvent casting method for wound dressing application. Optimization of hydrogel composition and analysis of wound dressing parameters such as stability and fluid uptake capacity (in the presence of water, saline and different pH solutions) has been studied. The result indicated that the PVA/beta-glucan hydrogel hold its structural integrity even at alkaline pH (pH~9) and upholds fluids four times of its original weight. Thus, the developed hydrogel is expected to be a promising candidate as wound dressing.

SIRT1 is a NAD-dependent protein deacetylase that participates in cellular regulation. The increased migration of fibroblasts is an important phenotype in fibroblast activation. The role of SIRT1 in cell migration remains controversial as to whether SIRT1 acts as an activator or suppressor of cell migration. Therefore, we have established the role of SIRT1 in the migration of human dermal fibroblasts and explored targets of SIRT1 during dermal fibroblast migration. SIRT1 and Cyr61 were expressed in human dermal fibroblasts and the stimulation with TGF-β further induced their expression. Treatment with resveratrol (RSV), a SIRT1 agonist, or overexpression of SIRT1 also promoted the expression Cyr61 in human dermal fibroblasts, whereas the inhibition of SIRT1 activity by nicotinamide or knockdown of SIRT1 decreased the level of Cyr61, as well as TGF-β or RSV-induced Cyr61 expression. Blocking of ERK signaling by PD98509 reduced the expression of Cyr61 induced by TGF-β or RSV. TGF-β, RSV, or SIRT1 overexpression enhanced β-catenin as well as Cyr61 expression. This stimulation was reduced by the Wnt inhibitor XAV939. RSV increased migration and nicotinamide attenuated RSV-induced migration of human dermal fibroblasts. Furthermore, SIRT1 overexpression promoted cell migration, whereas blocking Cyr61 attenuated SIRT1-stimulated migration of human dermal fibroblasts. SIRT1 increased cell migration by stimulating Cyr61 expression and the ERK and Wnt/β-catenin signaling. SIRT1-induced Cyr61 activity is very important for human dermal fibroblasts migration.

Manila clam Ruditapes philippinarum is one of the most important benthic animals in the coastal north Pacific region, where clam populations have been mixed genetically through trade and aquaculture activities. Accordingly, identification of the genetically different clam populations has become one of the most important issues to manage interbreeding of the local and introduced clam populations. To identify genetically different populations of clam populations, we developed 11 expressed sequence tag (EST)-microsatellite loci (i.e., simple sequence repeat, SSR) from 1,128 clam hemocyte cDNA clones challenged by the protozoan parasite Perkinsus olseni. Genotype analysis using the markers developed in this study demonstrated that clams from a tidal flat on the west coast contained 6 to 19 alleles per locus, and a population from Jeju Island had 4 to 20 alleles per locus. The expected heterozygosity of the 2 clam populations ranged from 0.472 to 0.919 for clams from the west coast, and 0.494 to 0.919 for clams from Jeju Island, respectively. Among the 11 loci discovered in this study, 7 loci significantly deviated from the Hardy-Weinberg equilibrium after Bonferroni correction. The 5 loci developed in this study also successfully amplified the SSRs of R. variegatus, a clam species taxonomically very close to R. philippinarum, from Hong Kong and Jeju Island. We believe that the 11 novel polymorphic SSR developed in this study can be utilized successfully in Manila clam genetic diversity analysis, as well as in genetic discrimination of different clam populations.

Epithelial to mesenchymal transition (EMT) is a complex biological event, wherein polarized epithelial cells lose their integrity resulting in a mesenchymal phenotype with enhanced motility, a phenomenon known as metastasis. However, the underlying mechanisms of EMT are still poorly understood in cervical carcinomas. In this study, we investigated the molecular signalling events responsible for the effect of TGF-β, a potent inducer of EMT, on HeLa cervical cancer cells. We observed that TGF-β treatment (5 ng/mL) upregulates the expression of EMT-associated transcription factors such as Snail and Slug and downregulates the expression of epithelial markers such as ZO-1 and E-cadherin. Furthermore, treatment with TGF-β activates both Smad-dependent and Smad-independent signaling pathways, which subsides upon addition of Diphenyleneiodonium (DPI), a potent ROS inhibitor that inhibits NADPH oxidase (NOX). TGF-β treatment enhanced cellular migration and invasion ability was diminished in the presence of ROS inhibitors. In addition, we also observed that ROS-mediated, TGF-β-induced EMT progression was inhibited using therapeutic candidates that target the key signal transduction mediators, including PI3K/AKT, ERK, and P38/MAPK. Accordingly, we demonstrated the involvement of redox biology (NOX2 and NOX4 mediate migration and invasion) in TGF-β-mediated EMT advancement and explored suitable therapeutic interventions.

Additional file 1: Fig. S1. Transcription factors not involved in IL-6 induced Cyr61 protein synthesis. RA-FLSs transfected with either small interfering RNA (NR4A1 or ATF3) or siNC (control) (20 pmol/L) stimulated by IL-6 (20 ng/mL) for 2 h. Data are representative of at least three independent experiments. Fig. S2. Expression of IL-6 enhanced by Cyr61 secretion. The mRNA level of IL-6 stimulated by Cyr61 protein (100 ng/mL) for 2 h as determined via real time polymerase chain reaction. Values are means (± standard deviation) of at least three independent experiments. **p

Western diet, rich in carbohydrates and fat, is said to be a major factor underlying metabolic syndrome. Interventions with prebiotics, the key modulators of the gut microbiota, have paramount impact on host-associated metabolic disorders. Herein, we investigated the effect of fungus-derived (1,3)/(1,6)-β-glucan, a highly soluble dietary fiber, on high-fat diet (HFD)-induced metabolic distress.Male C57BL/6 J mice were fed with different diet groups (n = 11): control diet, HFD, 3 g/kg or 5 g/kg of β-glucan-incorporated HFD. At the end of experimental study period (12th week), body weight, feces weight and fecal moisture content were observed. Further, colonic motility was measured using activated charcoal meal study. Proteins extracted from liver and intestine tissues were subjected to western blot technique. Paraffin-embedded intestinal tissues were sectioned for histochemical [Periodic acid-Schiff (PAS) and Alcian blue (AB) staining] analysis. Fecal microbiota analysis was performed using MOTHUR bioinformatic software.β-glucan consumption exhibited anti-obesity property in mice groups fed with HFD. In addition, β-glucan ameliorated HFD-induced hepatic stress, colonic motility and intestinal atrophy (reduction in colon length, goblet cells, and mucosal layer thickness). Further, β-glucan incorporation shifted bacterial community by increasing butyrate-producing bacteria such as Anaerostipes, Coprobacillus, and Roseburia and decreasing reportedly obesity-associated bacteria such as Parabacteroides and Lactococcus.Altogether, the outcomes of this present pre-clinical animal study show β-glucan to be a promising therapeutic candidate in the treatment of HFD-induced metabolic distress. Further comprehensive research has to be conducted to brace its clinical relevance, reproducibility and efficacy for aiding human health.

Reactive oxygen species (ROS) regulate cell fate, although signaling molecules that regulate ROS hormesis remain unclear. Here we show that transmembrane 4 L six family member 5 (TM4SF5) in lung epithelial cells induced the alternatively spliced CD44v8-10 variant via an inverse ZEB2/epithelial splicing regulatory proteins (ESRPs) linkage. TM4SF5 formed complexes with the cystine/glutamate antiporter system via TM4SF5- and CD44v8-10-dependent CD98hc plasma-membrane enrichment. Dynamic TM4SF5 binding to CD98hc required CD44v8-10 under ROS-generating inflammatory conditions. TM4SF5 and CD44v8-10 upregulated cystine/glutamate antiporter activity and intracellular glutathione levels, leading to ROS modulation for cell survival. Tm4sf5-null mice exhibited attenuated bleomycin-induced pulmonary fibrosis with lower CD44v8-10 and ESRPs levels than wild-type mice. Primary mouse alveolar epithelial cells (AECs) revealed type II AECs (AECII), but not type I, to adapt the TM4SF5-mediated characteristics, suggesting TM4SF5-mediated AECII survival following AECI injury during idiopathic pulmonary fibrosis (IPF). Thus, the TM4SF5-mediated CD44v8-10 splice variant could be targeted against IPF.

Western diet, rich in carbohydrates and fat, is said to be a major factor underlying metabolic syndrome. Interventions with prebiotics, the key modulators of the gut microbiota, have paramount impact on host-associated metabolic disorders. Herein, we investigated the effect of fungus-derived (1,3)/(1,6)-β-glucan, a highly soluble dietary fiber, on high-fat diet (HFD)-induced metabolic distress. Male C57BL/6 J mice were fed with different diet groups (n = 11): control diet, HFD, 3 g/kg or 5 g/kg of β-glucan-incorporated HFD. At the end of experimental study period (12th week), body weight, feces weight and fecal moisture content were observed. Further, colonic motility was measured using activated charcoal meal study. Proteins extracted from liver and intestine tissues were subjected to western blot technique. Paraffin-embedded intestinal tissues were sectioned for histochemical [Periodic acid-Schiff (PAS) and Alcian blue (AB) staining] analysis. Fecal microbiota analysis was performed using MOTHUR bioinformatic software. β-glucan consumption exhibited anti-obesity property in mice groups fed with HFD. In addition, β-glucan ameliorated HFD-induced hepatic stress, colonic motility and intestinal atrophy (reduction in colon length, goblet cells, and mucosal layer thickness). Further, β-glucan incorporation shifted bacterial community by increasing butyrate-producing bacteria such as Anaerostipes, Coprobacillus, and Roseburia and decreasing reportedly obesity-associated bacteria such as Parabacteroides and Lactococcus. Altogether, the outcomes of this present pre-clinical animal study show β-glucan to be a promising therapeutic candidate in the treatment of HFD-induced metabolic distress. Further comprehensive research has to be conducted to brace its clinical relevance, reproducibility and efficacy for aiding human health.

Purpose One of the features in cancer development is the migration of cancer cells to form metastatic lesions. CYR61 protein promotes migration and the epithelial-mesenchymal transition in several cancer cell types. Evidence suggests that CYR61 and dexamethasone are relevant to colorectal cancer. However, relationships between them and colorectal cancer are still unclear. Understanding the molecular mechanism of colorectal cancer progression related with CYR61 and dexamethasone, which is widely used for combination chemotherapy, is necessary for improved therapy. Materials and methods We used colorectal cancer cells, HCT116, co-treated with transforming growth factor β1 (TGF-β1) and dexamethasone to examine the inhibitory migration effect of dexamethasone by migratory assay. Alternatively, both migratory pathways, expression of AKT and ERK, and the target factor CYR61 was also tested by co-treatment with TGF-β1 and dexamethasone. Results We report that dexamethasone significantly inhibited TGF-β1-induced cell migration, without affecting cell proliferation. Importantly, we observed that TGF-β1 promoted the epithelial-mesenchymal transition process and that dexamethasone co-treatment abolished this effect. ERK and AKT signaling pathways were found to mediate TGF-β1-induced migration, which was inhibited by dexamethasone. In addition, TGF-β1 treatment induced CYR61 expression whereas dexamethasone reduced it. These observations were compatible with the modulation of migration observed following treatment of HCT116 cells with human recombinant CYR61 and anti-CYR61 antibody. Our results also indicated that TGF-β1 enhanced collagen I and reduced matrix metalloproteinase 1 expression, which was reversed by dexamethasone treatment. Conclusion These findings suggested that dexamethasone inhibits AKT and ERK phosphorylation, leading to decreased CYR61 expression, which in turn blocks TGF-β1-induced migration.

Hepatitis B virus infection can cause hepatic fibrosis leading to cirrhosis and hepatocellular carcinoma. However the mechanism remains poorly understood. In this study, we found that Hepatitis B virus X-protein (HBx) increases vimentin, fibronectin, slug, snail and NOX4 expression. Because NOX4- mediated reactive oxygen species can increase slug and snail, which can induce fibrosis, HBx may be a key regulator of hepatic fibrosis development via NOX4 induction.

Cutaneous wound repair is an intricate process whereby the skin reprograms itself after injury. In the mid-phase of wound repair, the proliferation, migration, and differentiation of cells are the major mechanisms to lead remodeling. We investigated the effect of BMM ((1E,2E)-1,2-bis((6-bromo-2H-chromen-3-yl)methylene)hydrazine), a novel synthetic material, on the migration and viability of keratinocytes or fibroblasts using the in vitro scratch woundhealing, electric cell-substrate imedance sensing (ECIS), invasion, and MTT assays. Cell migration-related factors were analyzed using western blot, and we found that treatment with BMM stimulated the EMT pathway and focal adhesion kinase (FAK)/Src signaling. Differentiation of HaCaT keratinocyte and fibroblast cells was also stimulated by BMM and specifically, NOX2/4 contributed to the activation of fibroblasts for wound healing. Furthermore, BMM treated HaCaT keratinocyte and fibroblast-co-cultured cells increased migration and differentiation. TGF-β and Cyr61 were also secreted to a greater extent than in single cultured cells. In vivo experiments showed that treatment with BMM promotes wound closure by promoting re-epithelialization. In this study, we demonstrated that a novel synthetic material, BMM, is capable of promoting wound healing via the stimulation of re-epithelialization in the epidermis and the activation of fibroblasts in the dermis, in particular, via the acceleration of the interaction between the epidermis and dermis.

Camphor ((1R)-1,7,7-trimethylbicyclo[2.2.1]heptan-2-one), a bicyclic monoterpene, is one of the major constituents of essential oils from various herbs such as rosemary, lavender, and sage. In this study, we investigated the beneficial effects of camphor as a botanical ingredient in cosmetics. Camphor induced the proliferation of human primary dermal fibroblasts in a dose-dependent manner via the PI3K/AKT and ERK signaling pathways. Camphor attenuated the elevation of senescence associated with β-galactosidase (SA-β-gal) activity. Elastase activity decreased, while the total amount of collagen increased, in a dose- and time-dependent manner in human primary dermal fibroblasts treated with camphor. Camphor induced the expression of collagen IA, collagen IIIA, collagen IVA, and elastin in human primary dermal fibroblasts. In addition, posttreatment with 26 and 52 mM camphor for 2 weeks led to a significant reduction in the expression of MMP1 but increases in the expression of collagen IA, IIIA, and elastin in mouse skin exposed to UV for 4 weeks. These posttreatments also reduced the depths of the epidermis and subcutaneous fat layer in UV-exposed mouse skin. Taken together, these findings suggest camphor to be a potent wound healing and antiwrinkle agent with considerable potential for use in cosmeceuticals.

Glycitin is a soy isoflavone that exhibits antioxidant, antiallergic, and anti-osteoporosis activities. We investigated the effects of glycitin on dermal fibroblast proliferation and migration. Treatment of primary dermal fibroblasts with glycitin increased cell proliferation and migration. In addition, treatment with 20 μM glycitin for 24 h induced the synthesis of collagen type I and type III at both the mRNA and protein levels. Fibronectin was also increased by 20% after treatment. Matrix metalloproteinase-1 collagenase was decreased in the media after 24-h incubation with glycitin, and the synthesis of transforming growth factor-beta (TGF-β) mRNA increased approximately twofold in cells following glycitin treatment. Phosphorylation of Smad2 and Smad3 increased after 1 h of glycitin treatment, and phosphorylation continued for 24 h. Furthermore, the phosphorylated form of AKT was increased in glycitin-treated cells after 3 h and remained higher for 24 h. Thus, glycitin treatment produces anti-aging effects including increased total collagen in the culture media, decreased elastase, and decreased β-galactosidase. Together, these results indicate that glycitin stimulates TGF-β secretion, and the subsequent autocrine actions of TGF-β induce proliferation of fibroblasts, ultimately protecting skin cells from aging and wrinkling. Copyright © 2015 John Wiley & Sons, Ltd.

A polyclonal antibody specific to an egg protein of Suminoe oyster Crassostrea ariakensis was previously developed in our laboratory to assess the reproductive life cycle of the oyster. The present study was undertaken to investigate vitellin of C. ariakensis (CAVt). Vitellin is an essential component of egg proteins in marine invertebrates as it provides energy and nutrients to the embryo and larvae. CAVt was purified from eggs of the oyster using ammonium sulfate precipitation followed by affinity chromatography with Concanavalin A-agarose. Native polyacrylamide gel electrophoresis (PAGE) and sodium dodecyl sulfate PAGE showed that CAVt is a high molecular weight [532 kiloDaltons (kDa)] protein, with multiple subunits. Similar to other vitellin proteins, it is a phospholipoglycoprotein composed of phospholipids (12.06%), carbohydrates (mannose, 10.08% or glucose, 9.84%), and alkali-labile phosphates (4.16%). Affinity chromatography, enzyme-linked immunosorbent aasay (ELISA) and western blot analysis revealed that CAVt is only present in the ovary, and two subunits of CAVt (72 and 35 kDa) are believed to be incorporated from the hemolymph into the oocyte. The antibody specific to CAVt (anti-CAVt), raised in rabbit, strongly cross reacted with the egg proteins of oyster species and scallops, suggesting that the antigenic epitopes are highly conserved among species. Our results suggest that the anti-CAVt antibody can be used to develop a tool similar to ELISA or western blotting for investigation of the effect of microorganisms on reproduction as well as the effect of chemicals on the endocrine system in C. ariakensis.

Background Peroxisome proliferator-activated receptor gamma (PPARγ) agonist has anti-inflammatory properties, which has known to reduce inflammatory cytokine production in RA. Cysteine-rich angiogenic inducer 61 (Cyr61) is associated with diseases related to chronic inflammation. Cyr61, pro-inflammatory factor, has been shown to be increased in the synovial tissues of patients with RA. However the action mechanisms between Cyr61 and PPARγ are unknown. To determine the effects of peroxisome proliferator–activated receptorγ (PPARγ) agonists on tumour necrosis factor alpha (TNF-α)- induced of FLS invasiveness phenotype and Cystein -rich angiogenic inducer 61 (Cyr61) in Rheumatoid arthritis Fibroblast-like synoviocytes (RA-FLS). Materials and methods FLS were cultured with TNF-α and Cyr61 in the presence or absence of PPARγ agonists. MMPs and CYR61 expression levels in the RA-FLS and culture medium were measured reverse transcriptase–polymerase chain reaction (RT-PCR) and western blotting. The migration and invasive phenotype of RA-FLS were determined by a scratch wound healing assay and the Boyden chamber assay. Results Cyr61 protein was expressed on RA-FLS, and its expression was increased by TNFα. Moreover, Cyr61 directly promoted RA-FLS migration (p Conclusion Our result show that PPARγ agonist may have beneficial effects on migration and invasion of RA-FLS via down-regulation of Cyr61. Therefore, PPARγ agonist could be a potential treatment of RA.

Background SIRT1 is a NAD-dependent protein deacetylase that participate in cellular regulation. The increased migration of fibroblast is important phenotype in fibroblast activation. The role of SIRT1 in cell migration remains controversial whether SIRT1 act as an activator or suppressor for cell migration. We have therefore established the role of SIRT1 in human dermal fibroblast migration and explored target of SIRT1 for dermal fibroblast migration. Results SIRT1 and Cyr61 were expressed in human dermal fibroblasts and the stimulation of TGF-β further induced expression SIRT1 and Cyr61. Treatment of resveratol (RSV), SIRT1 agonist or overexpression of SIRT1 also promoted expression of SIRT1 and Cyr61 in human dermal fibroblasts, whereas inhibition of SIRT1 activity by nicotinamide or knock down of SIRT1 down-regulated Cyr61 basal level as well as TGF-β or RSV-induced Cyr61expression. Blocking of ERK signalling by PD98509 reduced TGF-β or RSV-induced Cyr61 expression. TGF-β, RSV or SIRT1 vector enhanced β-catenin as well as Cyr61 expression. This stimulation was reduced by Wnt inhibitor, XAV939. RSV increased migration and nicotinamide attenuated RSV-induced migration of human dermal fibroblasts. Furthermore, SIRT1 overexpression promoted cell migration whereas blocking Cyr61 attenuated SIRT1-stimulated migration of human dermal fibroblasts. Conclusion SIRT1 increased cell migration by stimulated Cyr61 expression for their target through the ERK and Wnt/β-catenin signalling. SIRT1-induced Cyr61 production is very important for human dermal fibroblasts migration.

Accumulating evidence has displayed that targeting cancer stem cells (CSCs) is a very promising way for anti-cancer therapies. 2',3'-Dimethoxyflavanone (2',3'-DMF) showed the most potent toxicity of a group of 42 flavonoids tested in MCF-7-SC breast cancer stem cells. 2',3'-DMF triggered intrinsic and extrinsic apoptosis by stimulating the cleavage of PARP and the activation of caspase-9, -8, and -3. Interestingly, 2',3'-DMF induces a dramatic increase in the conversion of LC3, a well-known marker for autophagy. However, acidic vesicular organelles (AVOs), one of the autophagic flux markers were not detected. Co-treatment with chloroquine, the lysosomal inhibitor that blocks autophagic degradation did not show any change in the degree of LC3 conversion, implying that LC3 could play a role in the non-autophagic cell death of MCF-7-SC. We found that 2',3'-DMF induces the ubiquitination of caspase-8, this resulted in an interaction between caspase-8 and LC3, which led to the aggregation and activation of caspase-8. Co-treating cells with 2',3'-DMF and 3-methyladenine, an inhibitor of LC3 lipidation, reduced the activation of caspase-8. These findings provide novel insights into the anti-cancer effects of 2',3'-DMF in breast cancer stem cells by revealing that it induced apoptosis in accompany with the activation of caspase-8 mediated by LC3 conversion.

Hepatic stellate cells (HSCs) are the main extra cellular matrix-producing cells in the liver. Several reports have indicated that activated HSCs are involved in hepatic carcinogenesis by way of transforming growth factor β (TGF-β) secretion. This study aimed to investigate the effects of TGF-β, derived from HSCs activated by the chronic hepatitis B virus x protein (HBx), on the transdifferentiation of hepatocytes into hepatocarcinoma cells. Normal hepatocytes (the Chang liver cell line) were treated with a low concentration of TGF-β for 2 weeks, after which cell cycle- and cell signaling- related protein expression was analyzed. Lon-term treatment of TGF-β clearly induced the proliferation and the expression of cancer signaling proteins in the Chang cell line. TGF-β treatment also increased the expression of c-Jun N-terminal kinase (JNK) and c-Myc, indicating that induction of the JNK/pSmad3/c-Myc oncogenic signaling pathway is involved in hepatocyte transformation. Similar results were observed after culturing Chang cells with conditioned media derived from the activated LX-2 hepatic stellate cell line, suggesting that TGF-β paracrine effects are involved in the transformation of hepatocyte cells into hepatocarcinoma cells. Immunohistochemical results showed that the livers from HBx transgenic mice were composed of more activated HSCs and produced more TGF-β compared with those from normal mice. The TGF-β secreted from HBx-infected HSCs might induce transdifferentiation of hepatocytes into hepatocarcinoma, which is the fact that suggested a potential knowledge on liver cancer inhibition.

Exposure of strong and repeated UV on the skin leads to skin aging, characterized with wrinkling, sagging, dyspigmentation, and laxity. Numerous studies revealed that Matrix metalloproteinases are related to skin aging and functions as degrading enzyme of various types of collagen. Here, we attempted to evaluate the effectiveness of glycitin (4′-hydroxy-6-methoxyisoflavone- 7-d-glucoside) on skin aging and mechanisms of action in UV-irradiated human dermal fibroblasts. Especially we focused on the expression of Matrix metalloproteinase-1 (MMP-1), which degrades procollagen type-I in dermis, by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, Western blot, and reverse transcription polymerase chain reaction in cell lysates or media. Our results showed that glycitin increased the viability of human dermal fibroblast and alleviated MMP-1 expression caused by UV irradiation. In addition, synthesis of type-I collagen was increased and UV-induced phosphorylation of ERK/JNK/p38 was decreased in dose-dependent manners. Taken together, we demonstrated that treatment with glycitein have a protective effect on skin aging by inhibiting of MMP-1 and increasing of collagen through ERK/JNK/P38 down-regulation, which may be mediated by the inhibition of ERK, JNK, and p38 mitogen-activated protein kinases. We suggest that glycitin is a potential agent for the treatment of skin ageing.

The epithelial–mesenchymal transition (EMT) is a critical biological process characterized by morphological and behavioral changes in cells. The regulatory and signaling mechanisms of both developmental and pathological EMT have been investigated. Reactive oxygen species (ROS) play a role in early EMT, but the exact mechanism by which ROS are involved is unclear. We investigated ROS-mediated EMT in human HeLa cells. Transforming growth factor beta (TGF-β) treatments lead to dramatic NADPH oxidase 2 (NOX2) inductions in HeLa cells; antioxidant treatment prevented TGF-β-driven EMT. Over-expression of the p40phox subunit (NCF4) led to activation of the NOX2 complex and ROS production. We showed that NOX2 and NOX5 mRNA was increased, along with increased expression of several matrix metalloproteinases (MMPs) in response to NCF4 expression. Moreover, these changes were reversible upon ROS scavenging. Down-regulation of E-cadherin and up-regulation of Snail, Slug and vimentin occurred at the transcriptional level. We also showed that new EMT regulator, YB-1 is a downstream target in ROS-induced EMT. Together, these data suggest that ROS switching is necessary for increased EMT but is not required for the morphological changes that accompany EMT.

Flavonoids are major active ingredients in plants and are considered components of food that provide medical or health benefits. They have diversified structures and have effects on human health, including wound healing induction. More than a hundred flavonoids were screened for HaCaT keratinocytes cellular migration measurements and the relationships between their structural properties and the effects promoting cellular migration were examined. Here, among flavonoids used in the previous structure-activity relationship calculations, 4′,6,7-trimethoxyisoflavone (TMF) was one of the compounds showing the best activity, so that its molecular mechanism of the wound healing effect on HaCaT keratinocytes was investigated in more detail. Our data revealed that TMF increased the wound healing rate, but not the proliferation rate, in a dose-dependent manner. Treatment of keratinocytes with TMF influenced signaling pathways, affecting the phosphorylation of AKT and ERK in a time-dependent manner. TMF also induced the cell–cell adhesion protein E-cadherin, which is essential for promoting collective cell migration. Furthermore, the TMF treatment group also showed higher ROS and NOX2 transcriptional and protein levels. Correlating with matrix metalloproteinase induction by TMF, levels of extracellular matrix proteins such as collagens I and III were significantly lower in the treatment group. To confirm that the effects of TMF occur through the NOX2 pathway, we co-treated cells with TMF plus an NADPH inhibitor (DPI) or a ROS scavenger (NAC). Western blotting revealed that DPI and NAC attenuated the effect of TMF, suggesting that TMF induces ROS through the NOX2 pathway and regulates keratinocyte migration. In summary, TMF promotes wound healing through NOX2 induction, which leads to collective migration and MMP activation.

The estimated prevalence of chronic granulomatous disease (CGD) is between 1 in 200,000 and 1 in 250,000 individuals,with variable occurrence in different countries. According to a collation by the Korean College of Pediatric Clinical Immunology,the prevalence of CGD in Korea was 0.9 in 1,000,000 individuals. Although most regions of Korea had similar prevalence ofCGD, the prevalence of CGD on Jeju Island was a surprising 20.7 in 1,000,000 individuals. Previously, we demonstrated that allCGD patients on Jeju Island had an identical mutation in the CYBA gene. We hypothesized that the high prevalence of CGD onJeju Island is associated with an identical mutation inherited from a common ancestor or proband. In this study, we developedhighly sensitive assay using mutation-specific primers to detect the unique mutation in the CYBA gene on Jeju Island. Weinvestigated the frequency of subjects carrying the unique mutated allele in CYBA gene. Circa 700 subjects from Seogwipo City(~0.5% of the population of Seogwipo City) were enrolled, 1.1% of whom had the CYBA mutated allele. A significant differencewas observed between the expected and calculated numbers in the population of Seogwipo City (~2% of the population) byHardy?Weinberg equilibrium. Further studies are necessary to elucidate the frequency at which this mutant allele occurs in thepopulation of Jeju Island.

In this study, we investigated the anti-inflammatory and anti-allergic effects of Stichopus japonicus extract. The anti-inflammatory effect of S. japonicus was evaluated using the Griess reaction to evaluate nitric oxide (NO) release and reverse transcriptase-PCR (RT-PCR) to determine interleukin (IL)-6 and tumor necrosis factor-α (TNF-α) mRNA expression levels in LPS-stimulated RAW 264.7 murine macrophages. The anti-allergic activity of S. japonicus was investigated by performing β-hexosaminidase assay on antigen-stimulated RBL-2H3 rat mast cell lines and IL-4 mRNA expression. Our data revealed that S. japonicas water fraction (SJW) inhibited NO release without cytotoxicity in LPS-stimulated Raw 264.7 cells. The levels of IL-6 and TNF-α mRNA reduced following SJW treatment. In addition, SJW inhibited antigen-induced degranulation (e.g., IC50 value of 658 μg/mL) as well as IL-4 mRNA expression. Our results suggest that SJW possesses anti-inflammatory and anti-allergic effects.