Viability Of Epithelial Cells Research Articles

Curcumin induced oxidative stress attenuation by N-acetylcysteine co-treatment: a fibroblast and epithelial cell in-vitro study in idiopathic pulmonary fibrosis

BackgroundIdiopathic Pulmonary Fibrosis (IPF) is a fatal lung disease of unknown etiology with only two federally approved drug options. Given the complex molecular pathogenesis of IPF involving multiple cell types and multiple pathways, we explore the effects of a potential antifibrotic and antioxidant drug combination. Curcumin is a polyphenolic compound derived from turmeric with significant biological activity including a potential antifibrotic capacity. N-acetylcysteine (NAC) is a precursor to the antioxidant glutathione. To advance our understanding of these molecules, and to identify a clinical application, we present a small number of focused experiments that interrogates the effect of curcumin and NAC on pathways relevant to IPF in both fibroblasts and epithelial cells.MethodsPrimary epithelial cell and fibroblasts isolated from patients with IPF were challenged with a combination treatment of NAC and curcumin. Evaluation of the antifibrotic potential and effect on oxidative stress was performed through QPCR gene expression analysis and functional assays including scratch tests, viability assays, and measurement of induced reactive oxygen species.ResultsWe demonstrate that curcumin alone does have antifibrotic potential, but that effect is accompanied by proapoptotic increases in oxidative stress. Coupled with this, we find that NAC alone can reduce oxidative stress, but that epithelial cell viability is decreased through this treatment. However, co-administration of these two molecules decreases oxidative stress and maintains high cell viability in both cell types. In addition, this co-treatment maintains an antifibrotic potential.ConclusionsThese findings suggest a novel application for these molecules in IPF and encourage further exploration of this potential therapeutic approach.

Analysis of lacrimal gland derived mesenchymal stem cell secretome and its impact on epithelial cell survival

In situ regeneration of lacrimal gland (LG) tissue would be a promising approach to curatively treat dry eye disease (DED). Mesenchymal stem cells (MSC) exhibit therapeutic effects in a variety of pathological conditions and our group recently reported that their number increases in regenerating mouse LG. Since the therapeutic effects are suggested to arise from secreted trophic factors, the application of MSC-secreted proteins seems to be a promising approach to induce/enhance LG regeneration. Therefore, this study aims to optimize the isolation of murine LG-MSC and analyze their secretome to investigate its potential for LG epithelial cell survival in vitro. For optimization, LG-MSC were isolated by an explant technique or cell sorting and their secretome was investigated under normal and inflammatory conditions. Results showed that the secretome of MSC had beneficial effects on the viability of ethanol-damaged LG epithelial cells. Additional, Lipocalin-2, prosaposin, ras GTPase-activating protein-binding protein 1 (Rac1) and signal transducer and activator of transcription 1 (STAT1), proteins that were up-regulated under inflammatory conditions, further improved the cell survival of ethanol-damaged LG epithelial cells. Interestingly, recovery of cell viability was highest, when the cells were incubated with STAT1. Summarizing, this study identified promising proteins for further studies on LG regeneration.

Terpinen-4-ol and carvacrol affect multi-species biofilm composition

The aim of this study was to investigate the cytotoxic activity and inhibitory effect of terpinen-4-ol (T4ol) and carvacrol against single- and multi-species biofilms. The toxicity of each compound was tested on oral keratinocytes and evaluated by XTT assay. Inhibition and eradication of single-species biofilms were analyzed by crystal violet assay and the effect on multi-species biofilm composition was evaluated by qPCR. T4ol and carvacrol did not affect the epithelial cell viability, in contrast to chlorhexidine, which showed a high cytotoxic effect. Inhibition and eradication of single-species biofilms treated with T4ol and carvacrol were observed. The same inhibitory effect was observed for multi-species biofilms, especially on periodontal pathogens. In conclusion, specific concentrations of T4ol and carvacrol without toxicity towards the epithelial cells reduced the numbers of periodontal pathogens in single- and multi-species biofilms.

Influence of polymeric precursors on the viability of human cells of yttrium aluminum borates nanoparticles doped with ytterbium ions

Abstract In the technological process of manufacturing, distribution and recycling of nanoparticles, is easy to spread them in the air. The respiratory system is the primary target for inhaled NPs. The authors discuss the influence of the complexing compounds used in the “button up” synthesis of inorganic nanoparticles, on the cells activity during in vitro studies. We decided to examine the impact of yttrium aluminum borates (YAB) nanoparticles doped with ytterbium ions synthesized from various polymeric precursors on the viability of human bronchial epithelial cells using the WST-8 assay, depending on the concentrations of these nanoparticles. The optically active YAB:Yb3+ nanoparticles were obtained by the sol–gel method. In our experiments three various hydrophilic polysaccharides and two different acids were used as polymeric complexing agents. Luminescent materials may be utilised in various fields, including medical diagnostic, however YAB:Yb3+ NPs are considered to be harmful or inert. We have tested those particles to determine the way their shape influences biological response. We show the effect of NPs suspension in biological media and their behavior under physiological conditions. In our work we demonstrate the influence of polymeric agents on the viability of bronchial cells, and consequently, on the probably inflammatory response. In the present study we report, that mannitol with citric acid favours the formation of fibrous particles while others polymeric agents, as salicylic acid with sorbitol favour spherical shape formation. Cellular responses to fibrous particles are different from these to spherical ones because bio-persistent fibers induce multiplication and possibility to obtain inflammasome. Particles in fiber form trigger the inflammation process, and thus can cause cell multiplication effect. We determined whether the polymeric precursors can have impact on the activation of the surface of NPs, and if they trigger interactions of NPs with cells. It turns out, that the issue of the toxicity of nanoparticles is very complex and should be approached comprehensively. Our results confirmed that stability, agglomeration, cytotoxicity of NPs and the multiplication of cells, depend on polymeric agents used in sol–gel synthesis. It had been shown that the method of nanoparticles synthesis has a significant impact on cellular viability and multiplication of cells. Preliminary studies on human bronchial cell demonstrated that the high viability and proliferation depends on the polymer precursors used in synthesis process. To describe the structure of the obtained NPs the following methods were used: Scanning Electron Microscopy, Transmission Electron Microscopy, Atomic Forces Microscopy, X-ray diffraction and Raman Spectroscopy.

Use of ceragenins as a potential treatment for urinary tract infections

BackgroundUrinary tract infections (UTIs) are one of the most common bacterial infections. High recurrence rates and the increasing antibiotic resistance among uropathogens constitute a large social and economic problem in current public health. We assumed that combination of treatment that includes the administration ceragenins (CSAs), will reinforce the effect of antimicrobial LL-37 peptide continuously produced by urinary tract epithelial cells. Such treatment might be an innovative approach to enhance innate antibacterial activity against multidrug-resistant E. coli.MethodsAntibacterial activity measured using killing assays. Biofilm formation was assessed using crystal violet staining. Viability of bacteria and bladder epithelial cells subjected to incubation with tested agents was determined using MTT assays. We investigated the effects of chosen molecules, both alone and in combinations against four clinical strains of E. coli, obtained from patients diagnosed with recurrent UTI.ResultsWe observed that the LL-37 peptide, whose concentration increases at sites of urinary infection, exerts increased bactericidal effect against E. coli when combined with ceragenins CSA-13 and CSA-131.ConclusionWe suggest that the employment of combination of natural peptide LL-37 with synthetic analogs might be a potential solution to treat urinary tract infections caused by drug-resistant bacteria.

Caffeic Acid Prevented LPS-Induced Injury of Primary Bovine Mammary Epithelial Cells through Inhibiting NF-κB and MAPK Activation.

In our previous study, lipopolysaccharide (LPS) significantly reduced the cell viability of primary bovine mammary epithelial cells (bMEC) leading to cell apoptosis, which were prevented by caffeic acid (CA) through inhibiting NF-κB activation and reducing proinflammatory cytokine expression. While the underlying mechanism remains unclear, here, we determined that LPS induced the extensive microstructural damage of bMEC, especially the mitochondria and endoplasmic reticulum. Then, the obvious reduction of mitochondrial membrane potential and expression changes of apoptosis-associated proteins (Bcl-2, Bax, and casepase-3) indicated that apoptosis signaling through the mitochondria should be responsible for the cell viability decrease. Next, the high-throughput cDNA sequencing (RNA-Seq) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were employed to verify that the MAPK and JAK-STAT signaling pathways also were the principal targets of LPS. Following, the critical proteins (ERK, JNK, p38, and c-jun) of the MAPK signaling pathways were activated, and the release of proinflammatory cytokines (TNF-α, IL-1β, IL-6, and IL-8) regulated by NF-κB and MAPKs was significantly increased, which can promote a cascade of inflammation that induces cell injury and apoptosis. Meanwhile, CA significantly inhibited the activation of MAPKs and the release of proinflammatory cytokines in a dose-dependent manner, which were similar to its effects on the NF-κB activation that we previously published. So we concluded that CA regulates the proteins located in the upstream of multiple cell signal pathways which can reduce the LPS-induced activation of NF-κB and MAPKs, thus weakening the inflammatory response and maintaining cell structure and function, which accordingly inhibit apoptosis.

Effects of benzalkonium chloride on cell viability, inflammatory response, and oxidative stress of human alveolar epithelial cells cultured in a dynamic culture condition

Recently, the importance of inhalation toxicity assessment increased due to recent humidifier disinfectant-associated deaths in children. Benzalkonium chloride (BAC) is currently used as a cationic surfactant and germicide in food industry processing lines and as a hand sanitizer. Animal models are mainly used as a method of evaluating the inhalation toxicity of a hazardous substance, but that approach requires considerable amounts of time and cost. As a replacement for animal experiments, in vitro cell culture can be used to assess toxicity. However, such culture does not reflect the natural microenvironment of the lung, particularly its dynamic nature. In this study, we simulated normal breathing levels (tidal volume 10%, 0.2 Hz) through surface elongation of an elastic membrane in a dynamic culture system. The low-cost dynamic system provided easy control of breathing rate during lung cell culture. We assessed the toxicity using different concentrations of BAC (0, 2, 5, 10, 20, and 40 μg/mL) under static and dynamic culture conditions. Following 24 h of exposure to BAC, cellular metabolic activity, cell membrane integrity, interleukin-8 (IL-8) and reactive oxygen species (ROS) levels, and the total amount of protein in cells were analyzed. Our results showed that significant differences in cellular metabolic activity, as well as IL-8 and ROS profiles, between static and dynamic cell growth conditions, following BAC exposure.

Frontline Science: Aggregated neutrophil extracellular traps prevent inflammation on the neutrophil-rich ocular surface.

Eye rheum is a physiological discharge, which accumulates at the medial angle of the healthy eye soon after opening in the morning. Microscopic evaluation of eye rheum revealed the presence of viable neutrophils, bacteria, epithelial cells, and particles, aggregated by neutrophil extracellular traps. We observed that in the evening, during eye closure, high C5a recruited neutrophils to the tear film and activated them. In this hypoxic area rich in CO2 , neutrophils fight microbial aggressors by degranulation. Immediately after eye opening, the microenvironment of the ocular surface changes, the milieu gets normoxic, and loss of CO2 induces subtle alkalinization of tear film. These conditions favored the formation of neutrophil extracellular traps (NETs) that initially covers the ocular surface and tend to aggregate by eyelid blinking. These aggregated neutrophil extracellular traps (aggNETs) are known as eye rheum and contain several viable neutrophils, epithelial cells, dust particles, and crystals packed together by NETs. Similar to aggNETs induced by monosodium urate crystals, the eye rheum shows a robust proteolytic activity that degraded inflammatory mediators before clinically overt inflammation occur. Finally, the eye rheum passively floats with the tear flow to the medial angle of the eye for disposal. We conclude that the aggNETs-based eye rheum promotes cleaning of the ocular surface and ameliorates the inflammation on the neutrophil-rich ocular surfaces.

Fluorometholone modulates gene expression of ocular surface mucins.

Mucins are vital to keep the ocular surface hydrated. Genes encoding for mucins contain a glucocorticoid response element. The purpose of this study was to evaluate the effect of fluorometholone, a glucocorticoid receptor agonist used in the management of dry eye, on the gene expression of conjunctival and corneal epithelial cell mucins. Stratified cultures of human conjunctival and corneal epithelial cells were exposed to 25, 50 and 100nM of fluorometholone alone or in presence of mifepristone, a glucocorticoid receptor antagonist. The mRNA was isolated from the cells and reverse transcribed to cDNA. The cDNA was used for quantification of gene expression of mucin (MUC) 1, 4, 16 and 19 using real-time PCR. Fluorometholone caused a dose- and time-dependent increase in the gene expression of MUC1, MUC4, MUC16 and MUC19 in the conjunctival as well as corneal epithelial cells. Mifepristone, a glucocorticoid receptor antagonist, inhibited fluorometholone-mediated increase in the gene expression of conjunctival and corneal mucins. At the tested concentration, neither fluorometholone nor mifepristone caused any notable changes in the cellular phenotype or viability of conjunctival and corneal epithelial cells. Fluorometholone increases the gene expression of MUC1, MUC4, MUC16 and MUC19 in the conjunctival and corneal epithelial cells through activation of glucocorticoid receptors. The increased expression of mucins can be an additional possible mechanism contributing to the beneficial effects of fluorometholone in dry eye in addition to its well-known anti-inflammatory effects.

Antibacterial activity against porcine respiratory bacterial pathogens and in vitro biocompatibility of essential oils.

Bacterial respiratory infections affecting pigs such as pneumonia, pleuropneumonia, and pleurisy, are a major health concern in the swine industry and are associated with important economic losses. This study aimed to investigate the antibacterial activities of essential oils against major swine respiratory pathogens with a view to developing a potential alternative to antibiotics. Their synergistic interactions with the bacteriocin nisin was also examined. Lastly, we assessed the in vitro biocompatibility of the most efficient essential oils using a pig tracheal epithelial cell line. Of the nine essential oils tested, those from cinnamon, thyme, and winter savory were the most active against Streptococcus suis, Actinobacillus pleuropneumoniae, Actinobacillus suis, Bordetella bronchiseptica, Haemophilus parasuis, and Pasteurella multocida, with minimum inhibitory concentrations and minimum bactericidal concentrations ranging from 0.01 to 0.156% (v/v). The main component found in cinnamon, thyme, and winter savory oils were cinnamaldehyde, thymol, and carvacrol, respectively. Treating pre-formed S. suis and A. pleuropneumoniae biofilms with thyme or winter savory oils significantly decreased biofilm viability. We also observed a synergistic growth inhibition of S. suis with mixtures of nisin and essential oils from thyme and winter savory. Concentrations of nisin and cinnamon, thyme and winter savory essential oils that were effective against bacterial pathogens had no effect on the viability of pig tracheal epithelial cells. The present study brought evidence that essential oils are potential antimicrobial agents against bacteria associated with porcine respiratory infections.

Determination of trypan blue efficacy in the mitigation of ex vivo canine PCO formation.

To determine whether trypan blue (TB) reduces canine lens epithelial cell (LEC) or corneal endothelial cell (CEC) viability in vitro; if cell death is noted, to subsequently evaluate the molecular mechanism. Cellular viability was determined using a lactate dehydrogenase (LDH) assay. In TB-treated LECs, caspase 3/7 activity was assessed to evaluate apoptosis; autophagy was evaluated using immunoblotting against LC3 and p62. To evaluate the effects of TB on ex vivo posterior capsule opacification (PCO), following mock cataract surgery, lens capsules were treated with TB and subsequently maintained in culture to determine LEC migration and proliferation. Following acute exposure, TB did not significantly reduce LEC or CEC viability at any of the concentrations tested. Increased caspase 3/7 activity was found in LEC cultures treated with TB for an extended period of time; no change in LC3 or p62 expression was noted. Ex vivo PCO formation was not significantly altered by TB treatment. Acute exposure to TB did not reduce LEC or CEC viability, and only longer exposure to TB was able to initiate apoptosis. Treatment with intraocular TB at the time of cataract surgery is likely safe to the CECs but will not prevent PCO formation.

Wood Smoke Disrupts Alveolar Epithelial Barrier Function through a p44/42 MAPK Signaling Pathway

BackgroundWood smoke (WS) contains many of the same toxic compounds as those found in cigarette smoke (CS). Although CS is a leading cause of respiratory disease, limited studies investigating the mechanisms involved in WS mediated respiratory disease have been undertaken. Exposure to WS particulate reduces epithelial cell viability and barrier function. Both CS and WS induce MAPK signaling and contribute to airway remodeling. With an increase in the size, duration, and number of forest fires annually, there is a need for a better understanding with regards to the acute and chronic effects of WS on respiratory health. Therefore, we undertook studies to evaluate the role of WS exposure on the function and remodeling of the alveolar epithelium.MethodsBirch wood was combusted and the smoke bubbled into PBS. The pH pf the resulting solution was adjusted to 7.1 and was considered 100% WS solution. Monolayers of A549 alveolar epithelial cells were exposed to increasing concentrations of WS for up to 48 hours. Cell viability was determined using MTT and trypan blue exclusion. Epithelial barrier function was monitored via electrical cell‐substrate impedance sensing (ECIS). Adhesion junctional structure was observed by immunofluorescent staining for E‐cadherin. Western blot was used to assess activation of MAPK signaling pathways.ResultsConcentrations of WS up to 10% had no effect on cell viability. WS promoted a concentration dependent reduction in epithelial barrier function after 24 hours which was maintained out to 48 hours. Cells not exposed to WS demonstrated distinct cobblestone E‐cadherin staining. Exposure to 2.5% WS diminished the cobblestone patterning after 24 hours and was completely lost after exposure to 5.0% WS. The wound healing capacity of cells after WS exposure was assessed using a scratch assay. After 48 hours, cells not exposed to WS had closed the scratched area by >50% whereas those cells exposed to 10% WS had closed the area by <20%. Exposure to WS induced phosphorylation of the p44/42, but not p38, MAPK signaling pathway. Pre‐treatment with the MAPK inhibitor U0126 prevented WS mediated epithelial barrier dysfunction. Additionally, U0126 preserved E‐cadherin cobblestone patterning even after WS exposure and had no effect on cell viability.ConclusionsExposure to WS disrupts alveolar epithelial adhesion that is partially regulated through the p44/42 MAPK signaling pathway. Pharmacological inhibition of p44/42 phosphorylation preserved epithelial barrier function, which may be as a result of maintained E‐cadherin‐mediated cell adhesion.Support or Funding InformationGrant in aide funding was provided by provided CIHR (DJG). MRZ and CTT are recipients of CIHR Fellowships. MRZ also received a MSFHR Trainee Award. CC is a Canada Research Chair and holds the Astra‐Zeneca Chair in Occupational and Environmental Lung Disease at the University of British Columbia.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Intracellular Degradation of Helicobacter pylori VacA Toxin as a Determinant of Gastric Epithelial Cell Viability.

Helicobacter pylori VacA is a secreted pore-forming toxin that induces cell vacuolation in vitro and contributes to the pathogenesis of gastric cancer and peptic ulcer disease. We observed that purified VacA has relatively little effect on the viability of AGS gastric epithelial cells, but the presence of exogenous weak bases such as ammonium chloride (NH4Cl) enhances the susceptibility of these cells to VacA-induced vacuolation and cell death. Therefore, we tested the hypothesis that NH4Cl augments VacA toxicity by altering the intracellular trafficking of VacA or inhibiting intracellular VacA degradation. We observed VacA colocalization with LAMP1- and LC3-positive vesicles in both the presence and absence of NH4Cl, indicating that NH4Cl does not alter VacA trafficking to lysosomes or autophagosomes. Conversely, we found that supplemental NH4Cl significantly increases the intracellular stability of VacA. By conducting experiments using chemical inhibitors, stable ATG5 knockdown cell lines, and ATG16L1 knockout cells (generated using CRISPR/Cas9), we show that VacA degradation is independent of autophagy and proteasome activity but dependent on lysosomal acidification. We conclude that weak bases like ammonia, potentially generated during H. pylori infection by urease and other enzymes, enhance VacA toxicity by inhibiting toxin degradation.

Arecoline suppresses epithelial cell viability through the Akt/mTOR signaling pathway via upregulation of PHLPP2

Arecoline, the major active ingredient of the betel nut, is involved in the pathogenesis of oral submucous fibrosis. However, the underlying mechanism of this pathogenesis remains unclear. In this study, we found that arecoline suppresses the cell proliferation of the HaCaT epithelial cell and induces cell cycle arrest at the G1/S phase with an IC50 of 50 μg/mL. Furthermore, we found that arecoline reduces the protein level of cyclin D1, but it has no effect on its mRNA level and protein stability, implying that arecoline may modulate the translation of cyclin D1. We also observed the downregulation of the Akt/mTOR signaling pathway after treatment with arecoline, which may be related to the translation of cyclin D1. RNA-seq analysis identified that PHLPP2, the direct upstream target of Akt, is significantly upregulated after arecoline treatment. siRNA-mediated knockdown of PHLPP2 recovered the phosphorylation state of Akt, as well as attenuated the effect of arecoline on cell viability. Thus, our study revealed the crucial role of PHLPP2 in arecoline-induced cell viability suppression.

Acids ‘generally recognized as safe’ affect morphology and biocompatibility of electrospun chitosan/polyethylene oxide nanofibers

Electrospinning of neat chitosan is currently achieved by using strong acids or organic solvents, which limits the use of chitosan nanofibers as biocompatible scaffolds for drug delivery and tissue engineering. The aim was to elucidate the effect of specific acids generally recognized as safe (GRAS) on the properties of electrospun chitosan-based nanofibers. Electrospinning chitosan in dilute acetic acid or succinic acid with polyethylene oxide resulted in white and separated nanofibers, whereas nanofibers electrospun in dilute citric acid were transparent and interconnected. Including succinic or citric acid in the spinning process induced disintegration of the fiber mat after four hours in water, and a concentration-dependent effect on epithelial cell viability. Chitosan nanofibers electrospun in acetic acid maintained their shape and fibrous structure after four hours in water, and showed no effect on cell viability. This study demonstrates that the choice of GRAS acid highly determines the properties of electrospun chitosan nanofibers.

A Simple Method to Determine Cytotoxicity of Water-Soluble Organic Compounds and Solid Particles from Biomass Combustion in Lung Cells in Vitro.

Adverse health effects of condensable organic compounds (COC) and potential secondary organic aerosols from wood combustion emissions are difficult to determine. Hence, available information is usually limited to a small number of specific applications. Therefore, we introduced a simple, fast, and economic method where water-soluble COC (WSCOC) and WSCOC together with water-soluble primary solid particles (WSpSP) from wood combustion were sampled and subsequently exposed to cultured human lung cells. Comparing the cell viability of H187 human epithelial lung cells from five combustion devices, operated at different combustion conditions, no, or only a minor, cytotoxicity of WSCOC is found for stationary conditions in a grate boiler, a log wood boiler, and a pellet boiler. All combustion conditions in a log wood stove and unfavorable conditions in the other devices induce, however, significant cytotoxicity (median lethal concentration LC50 5-17 mg/L). Furthermore, a significant correlation between CO and cytotoxicity was found ( R2 ∼ 0.8) suggesting that the simply measurable gas phase compound CO can be used as a first indicator for the potential harmfulness of wood combustion emissions. Samples containing WSCOC plus WSpSP show no additional cytotoxicity compared to samples with COC only, indicating that WSCOC exhibit much higher cytotoxicity than WSpSP.

Evaluation the Effect of Royal Jelly on the Growth of Two Members of Gut Microbiota; Bacteroides fragillis and Bacteroides thetaiotaomicron

Objective: In this study the effect of Royal jelly on the growth of two important members of Bacteroides spp.; Bacteroides fragilis and Bacteroides thetaiotaomicron, was evaluated. Also the physicochemical properties and cytotoxicity effects of Royal jelly on Caco-2 cell line as gastrointestinal epithelial cell model, assessed.
 Methods : Bacteria, Bacteroides fragilis and Bacteroides thetaiotaomicron were grown on brain heart infusion (BHI) broth medium supplemented with Royal jelly in 3 different concentrations (2.5, 5 and 10% v/v), both of the bacteria (1.5×108 cfu/mL) were inoculated to BHI broth contained Royal jelly in anaerobic condition. To calculate the bacterial optical density (OD), the absorbance was measured at 600 nm after an overnight. Also Caco-2 cells, was used to study the effects of Royal jelly on epithelial cell viability, and the Physicochemical properties consist of total proteins, polysaccharides, phenolic compounds, total lipids, ash and moisture by UV-VIS spectrophotometric and gravimetric methods were evaluated .
 Results: The growth of B. fragillis and B. thetaiotaomicron were increased by Royal jelly (2.5, 5 and 10% v/v concentrations) and the results indicated that Royal jelly increased the growth of bacteria in a dose dependent manner (p<0.001). In addition MTT assay showed more than 95% viability of Caco-2 cells treated with Royal jelly. The Iranian Royal jelly sample contains 59.01% water, 11.57% proteins, 12% lipids, 12.63 % polysaccharide and 5% mineral.
 Conclusion: The present study showed that Royal jelly has a potential effect in the preserving gut microbiota and it is suggested that Royal jelly as a complementary and alternative medicine can be used to treatment diseases are associated with gut microbiota- host interactions and immune regulating. Although we need to expand our knowledge by designing clinical trials to confirm the therapeutic effects of Royal jelly on gut microbiota modulation as a barrier function.

Chemopreventive properties of young green barley extracts in in vitro model of colon cancer

Young green barley is the most valuable source of nutrients and bioactive substances. It has a broad spectrum of health-promoting properties such as antioxidant, anti-inflammatory, hypoglycaemic, anti-depressant, anti-atherosclerotic and anticancer. The presented study is an attempt to extend this knowledge with particular emphasis on the possibility of using green barley in colon cancer prevention. Extracts were prepared on the basis of two commercial products: ground dried barley grass (YGB INT) and powder of young green barley juice (YGB GW). Their influence on colon epithelial cells (CCD841 CoN) viability and proliferation were analyzed by LDH and MTT assays. Anticancer properties of extracts were screened on colon cancer cell lines (LS180, HT-29) by MTT and BrdU assays. Changes in cells morphology induced by extracts were investigated after May-Grünwald-Giemsa staining. Tested extracts were not toxic against CCD841 CoN and did not affected their proliferation or morphology (LDH test, MTT test, microscopy observation). The MTT revealed that extracts significantly inhibited proliferation of colon cancer cells in a dose-dependent manner. Results of BrdU test confirmed antiproliferative properties of extracts, but opposite to MTT test, indicated YGB GW as a better anticancer agent. Light microscopy observation proved the data obtained from both MTT and BrdU tests and additionally suggested the ability of the extracts to induce necrosis in LS180 and HT-29 cells. The study demonstrated that YGB extracts specifically inhibit proliferation of colon cancer cells without any undesirable effect on colon epithelial cells. Obtained results will provide a rationele for the future development of dietary supplements which could be beneficial in colon cancer chemoprevention.

Limonin attenuates the stemness of cervical carcinoma cells by promoting YAP nuclear-cytoplasmic translocation

This work aimed to investigate the role and related mechanism of limonin in regulating the stemness of cervical carcinoma (CC) cells. In the present study, we constructed adriamycin-resistant CC cells and found that they exhibited greater stemness than parental cells. Additionally, limonin attenuated the stemness of CC cells that were resistant or sensitive to adriamycin, as evidenced by the decreases in spheroid formation capacity, stemness markers expression and ALDH1 activity, whereas limonin did not affect the viability of normal cervical epithelial cells. Furthermore, limonin enhanced adriamycin sensitivity and attenuated adriamycin resistance in CC cells. Mechanistically, the nuclear-cytoplasmic translocation of YAP, not TAZ, was promoted by limonin in CC cells. Additionally, YAP overexpression attenuated the inhibitory effects of limonin on CC cell stemness. Therefore, limonin can attenuate the stemness, and thus the chemoresistance, of CC cells by promoting the nuclear-cytoplasmic translocation of YAP.

RETRACTED: ANRIL acts as onco-lncRNA by regulation of microRNA-24/c-Myc, MEK/ERK and Wnt/β-catenin pathway in retinoblastoma.

This article has been retracted: please see Elsevier Policy on Article Withdrawal (http://www.elsevier.com/locate/withdrawalpolicy). This article has been retracted at the request of the Authors. Current research findings in the authors' laboratory are in conflict with previous experimental results published in this paper. It was found that the data reported in figures 4 and 5 were not able to be replicated and so therefore the authors have decided that it is best for the scientific record to retract this paper. All the authors agree with the retraction. The authors apologise for any confusion this paper might have caused readers.