Sleep duration and inflammatory mediator levels associated with long-term prognosis in temporomandibular disorders.

BACKGROUND: Sleep disturbance is a systemic symptom and at the same time a major modulating factor of temporomandibular disorders (TMD). Inflammation is known as a underlying mechanism involved in both poor sleep and increased pain. OBJECTIVE: The relationship between long-term clinical characteristics and hematologic biomarkers of hypothalamic-pituitary-adrenal axis activity and inflammation in TMD patients according to sleep duration was investigated to verify the possible role of sleep disturbance and systemic inflammation in TMD. MATERIALS AND METHODS: Inflammatory and stress mediator levels of venous blood samples were investigated in 63 female TMD patients along with comorbidity levels including stress, somatization, autonomic symptoms and sleep quality based on structured questionnaires. Differences in long-term clinical characteristics and hematologic variables following conservative treatment were analysed according to total sleep time as normal, short and long sleep groups. Also, clinical and hematologic indices related to favourable treatment response were sought out. RESULTS: Significantly less patients in the long sleep group reported pain on voluntary mandibular movement (p = .042) while depression (p = .043) and somatization levels (p = .002) were significantly higher in the short sleep group. Norepinephrine levels of the long sleep group were significantly lower than other groups. Decrease in pain intensity with treatment was smallest in the short sleep group. Erythrocyte sedimentation rate was associated with significant pain improvement at 3 months post-treatment and interleukin-1ß, -4, and -8 levels could predict favourable treatment response. CONCLUSION: Short sleep is associated with more comorbidities and unfavourable long-term treatment response in TMD which may be mediated by systemic inflammation. Effective management of sleep is necessary for successful TMD management.

On-site applicable diagnostic fluorescent probe for fire blight bacteria.

Fire blight is a representative plant infection that contaminates edible plants and causes socio-economic problems in agricultural and livestock industries globally. It is caused by the pathogen Erwinia amylovora (E. amylovora) creates lethal plant necrosis and spreads rapidly across plant organs. We newly disclose the fluorogenic probe B-1 for real-time on-site detection of fire blight bacteria for the first time. B-1 exhibited no emission signals but manifested bright emission properties in the presence of fire blight bacteria. Based on these features, fluorescence imaging of the fire blight bacteria and its real-time detection from the infected host plant tissues were conducted. The detection limit against E. amylovora was 102 CFU/mL, which had excellent sensitivity. The fluorogenic probe-based on-site diagnostic technology was supplemented by introducing a new portable UV device. This work holds enormous potential to be a new advanced tool for detecting fire blight in agricultural and livestock industries.

Porous silicon surface modification via a microwave-induced in situ cyclic disulfide (S-S) cleavage and Si-S bond formation.

Porous silicon (pSi) materials have gained a great deal of attention from various research fields, and their surface-functionalization is one of the critical points for their applications. In this study, a new surface modification method of Si-H-terminated pSi materials via microwave-induced Si-S bond formation is disclosed. The silicon hydride (Si-H) functionality on the pSi surface could react with the 5-membered cyclic disulfide (S-S) compound (DL-α-lipoic acid in this study) by microwave-induced in situ S-S bond cleavage and Si-S bond formation. This surface chemistry is fast responsive (<10 min) and more efficient than other methods such as vortexing, heating stirring, or ultrasonication. The reaction maintains the primary porous structure of pSi materials including pSi wafer, pSi rugate filer, and pSi nanoparticles. An additional functional group such as carboxylic acid is demonstrated to be readily introducible on the pSi surface for further applications. Overall, this study has successfully demonstrated the porous silicon surface modification via a microwave-induced in situ cyclic disulfide (S-S) cleavage and Si-S bond formation.

SIWV tetrapeptide and ROS-responsive prodrug conjugate for advanced glioblastoma therapy.

A new conjugate formulation, SIWV-PB-SN, based on glioblastoma (GBM)-homing SIWV tetrapeptide and an ROS-responsive prodrug is reported. SIWV-PB-SN selectively penetrates the GBM cells and releases anti-GBM drug (SN-38) via ROS-induced linker cleavage. This study presents a new insight for a more advanced therapeutic approach to overcoming GBM.

Development of a fluorescent nanoprobe based on an amphiphilic single-benzene-based fluorophore for lipid droplet detection and its practical applications.

Lipid droplets (LDs) are crucial biological organelles connected with metabolic pathways in biological systems and diseases. To monitor the locations and accumulation of LDs in lipid-related diseases, the development of a visualization tool for LDs has gained importance. In particular, LD visualization using fluorescent probes has gained attention. Herein, a new fluorescent nanoprobe, BMeS-Ali, is developed that can sense LDs based on an amphiphilic single benzene-based fluorophore (SBBF). BMeS-Ali consists of hydrophilic (-NH2) and hydrophobic (-C12H25) moieties and exists as a micelle nanostructure in aqueous media. BMeS-Ali has a weak fluorescence, but its emission was dramatically enhanced upon exposure to the LD components such as oleic acids (OA) by reassembling its nano-formulation. BMeS-Ali showed a selective LD staining ability and great biocompatibility in cells (cancer cells and stem cells). It also showed a practical sensing ability towards biologically derived lipids and can be applied to the visualization of human fingerprints. We found that the nanoprobe BMeS-Ali has significant potential to serve as a practical dye and sensor for lipids, especially for LD imaging in the biomedical research area and broader industrial applications.

Long-term evaluation of temporomandibular disorders in association with cytokine and autoantibody status in young women.

Temporomandibular disorders (TMD) is a chronic pain disease affecting 4-60% of general population. Its suggested etiology includes mechanical overloading to related structures, psychosocial factors, and genetic vulnerability. However, its pathogenesis is yet to be fully understood, especially in cases with a higher level of pain and more associated comorbidities. Recently chronic systemic inflammation and possible autoimmunity has been indicated in several pain conditions as the underlying mechanism of chronicity but this aspect has not been rigorously investigated in TMD. This article focuses on analyzing the levels of cytokines, chemokines, autoantibodies and nonspecific inflammatory markers and comparing their levels according to pain severity and duration in 66 female TMD patients in their 20 s and investigating their association with clinical indices of TMD and comorbidities. The high pain disability group showed decreased range of jaw function and more pain on palpation of capsule areas compared to the low pain disability group. Comorbidities such as anxiety and sleep disturbance were also significantly more prevalent. The level of IL-8 and IgG were significantly higher in the high pain disability group. IL-2, -8, -13, IFN- γ, RANTES, PGE2, and thrombopoietin levels showed a significant effect on indices reflecting jaw function, generalized pain intensity, and health related quality of life. Such results imply that longer pain duration and higher pain intensity is associated with higher levels of systemic inflammation suggesting the possible role of immunologic disturbance as an underlying factor of chronic TMD pain and warranting further investigation for its consideration in diagnosis and treatment.

Modelling and manufacturing of 3D-printed, patient-specific, and anthropomorphic gastric phantoms: a pilot study.

Interventional devices including intragastric balloons are widely used to treat obesity. This study aims to develop 3D-printed, patient-specific, and anthropomorphic gastric phantoms with mechanical properties similar to those of human stomach. Using computed tomography gastrography (CTG) images of three patients, gastric phantoms were modelled through shape registration to align the stomach shapes of three different phases. Shape accuracies of the original gastric models versus the 3D-printed phantoms were compared using landmark distances. The mechanical properties (elongation and tensile strength), number of silicone coatings (0, 2, and 8 times), and specimen hardness (50, 60, and 70 Shore A) of three materials (Agilus, Elastic, and Flexa) were evaluated. Registration accuracy was significantly lower between the arterial and portal phases (3.16 ± 0.80 mm) than that between the portal and delayed phases (8.92 ± 0.96 mm). The mean shape accuracy difference was less than 10 mm. The mean elongations and tensile strengths of the Agilus, Elastic, and Flexa were 264%, 145%, and 146% and 1.14, 1.59, and 2.15 MPa, respectively, and their mechanical properties differed significantly (all p < 0.05). Elongation and tensile strength assessments, CTG image registration and 3D printing resulted in highly realistic and patient-specific gastric phantoms with reasonable shape accuracies.

Safety and efficacy of a metal stent covered with a silicone membrane containing integrated silver particles in preventing biofilm and sludge formation in endoscopic drainage of malignant biliary obstruction: a phase 2 pilot study.

BACKGROUND AND AIMS: Membrane-covered self-expandable metal stents (SEMSs) have been developed to prolong the patency of stents by reducing tissue hyperplasia or tumor ingrowth. However, their effectiveness is attenuated by stent clogging as a result of biofilm formation on the inner surface of the membrane. The aim of this pilot study was to evaluate the efficacy and safety of SEMSs covered with a silicone membrane containing integrated silver particles (Ag-P) in malignant distal biliary obstruction. METHODS: Twenty-four patients who underwent SEMS placement because of malignant distal biliary obstruction were enrolled in this single-center pilot study. The main outcomes were technical success, clinical success, adverse events, stent patency, and survival. RESULTS: The technical and clinical success rates were 100% and 91.7% (22 of 24), respectively. The rates of early and late adverse events were 22.7% and 36.4%, respectively. The primary reintervention rate was 27.3% (6 of 22). Only 1 case involving stent malfunction was associated with sludge impaction. Median stent patency was 179 days. During follow-up, there were no serious adverse events or mortality related to the stents or Ag-P. Serum and urine silver concentrations before and after stent placement and at 32 weeks after placement did not differ. All serum and urine silver concentrations were <3 µg/L (3 ppb) and 5 µg/L (5 ppb), respectively. CONCLUSIONS: SEMSs covered with a silicone membrane containing integrated Ag-Ps may be effective and safe in malignant distal biliary obstruction. Stent dysfunction related to sludge impaction may be less frequent using this new stent. (Clinical Research Information Service identifier: KCT 0002310.).

Characterization and Differentiation of Circulating Blood Mesenchymal Stem Cells and the Role of Phosphatidylinositol 3-Kinase in Modulating the Adhesion.

Bone marrow mesenchymal stem cells (BM MSCs) can differentiate into multi-lineage tissues. However, obtaining BM MSCs by aspiration is difficult and can be painful; therefore peripheral blood (PB) MSCs might provide an easier alternative for clinical applications. Here, we show that circulating PB MSCs proliferate as efficiently as BM MSCs in the presence of extracellular matrix (ECM) and that differentiation potential into osteoblast in vitro and in vivo. Both BM MSCs and PB MSCs developed into new bone when subcutaneously transplanted into immune-compromised mice using hydroxyapatite/tricalcium phosphate as a carrier. Furthermore, LY294002 and Wortmannin blocked mesenchymal stem cell attachment in a dose-dependent manner, suggesting a role of phosphatidylinositol 3-kinase in MSC attachment. Our data showed that the growth of PB MSCs could be regulated by interaction with the ECM and that these cells could differentiate into osteoblasts, suggesting their potential for clinical applications.

Synergistic Effect of Adipose-Derived Stem Cells and Fat Graft on Wrinkles in Aged Mice.

BACKGROUND: The authors investigated the synergistic effects of adipose-derived stem cells and fat graft on skin wrinkles in a nude mouse model of chronologic aging. METHODS: After 50 weeks of chronologic aging, 44 female BALB/c nude mice were classified into four groups: (1) negative control, (2) mice injected subcutaneously with fat on the back skin (0.5 cm), (3) mice injected with adipose-derived stem cells (1 × 10 cells in 0.5 cm Hanks balanced salt solution), and (4) mice injected with both fat (0.5 cm) and adipose-derived stem cells (1 × 10 cells in 0.5 cm Hanks balanced salt solution). The degree of wrinkling was evaluated using replica analysis, and skin biopsies were performed after 4 weeks. The dermal thickness and density of collagen were determined. Type I procollagen and matrix metalloproteinase levels were determined using real-time polymerase chain reaction and Western blot analysis. Tropoelastin, fibrillin-1, and CD31 levels were evaluated using immunohistochemistry. RESULTS: Based on the total wrinkle area, there was significant wrinkle reduction in the fat-treated and adipose-derived stem cell with fat-treated groups. Type I procollagen mRNA and collagen levels were significantly higher in the adipose-derived stem cell with fat-treated group than in the adipose-derived stem cell-treated and the fat-treated groups. In addition, the adipose-derived stem cells with fat graft group exhibited significantly higher CD31 expression level than the adipose-derived stem cell-treated and the fat-treated groups. CONCLUSION: Both adipose-derived stem cells and fat graft have a wrinkle-reducing effect and synergistically affect collagen synthesis and neovascularization.

Patient-derived multicellular tumor spheroids towards optimized treatment for patients with hepatocellular carcinoma.

BACKGROUND: Hepatocellular carcinoma (HCC) is one of the most common malignant tumors worldwide and has poor prognosis. Specially, patients with HCC usually have poor tolerance of systemic chemotherapy, because HCCs develop from chronically damaged tissue that contains considerable inflammation, fibrosis, and cirrhosis. Since HCC exhibits highly heterogeneous molecular characteristics, a proper in vitro system is required for the study of HCC pathogenesis. To this end, we have established two new hepatitis B virus (HBV) DNA-secreting HCC cell lines from infected patients. METHODS: Based on these two new HCC cell lines, we have developed chemosensitivity assays for patient-derived multicellular tumor spheroids (MCTSs) in order to select optimized anti-cancer drugs to provide more informative data for clinical drug application. To monitor the effect of the interaction of cancer cells and stromal cells in MCTS, we used a 3D co-culture model with patient-derived HCC cells and stromal cells from human hepatic stellate cells, human fibroblasts, and human umbilical vein endothelial cells to facilitate screening for optimized cancer therapy. RESULTS: To validate our system, we performed a comparison of chemosensitivity of the three culture systems, which are monolayer culture system, tumor spheroids, and MCTSs of patient-derived cells, to sorafenib, 5-fluorouracil, and cisplatin, as these compounds are typically standard therapy for advanced HCC in South Korea. CONCLUSION: In summary, these findings suggest that the MCTS culture system is the best methodology for screening for optimized treatment for each patients with HCC, because tumor spheroids not only mirror the 3D cellular context of the tumors but also exhibit therapeutically relevant pathophysiological gradients and heterogeneity of in vivo tumors.

Surface Characterization and Human Stem Cell Behaviors of Zirconia Implant Disks Biomimetic-Treated in Simulated Body Fluid.

PURPOSE: This study investigated the effects of biomimetic deposition on a zirconia surface in simulated body fluid (SBF) and assessed the proliferation and differentiation of human bone marrow mesenchymal stem cells on the SBF-treated zirconia disks. MATERIALS AND METHODS: Corrected SBF was prepared according to Kokubo's recipe. Eighty yttrium oxide-stabilized tetragonal zirconia polycrystalline disks were prepared and divided into two groups: (1) the test group with SBF-treated disks and (2) the control group with nontreated disks. Zirconia disks were soaked in SBF for 1, 4, 7, and 14 days at 36.5°C, and the hydroxyapatite (HA) precipitation was verified by analyzing the surface morphology. For more in-depth validation of HA formation, the surface roughness, composition, and crystallization of the 7-day treated disks were analyzed. Human bone marrow mesenchymal stem cells were used to further evaluate cell proliferation, alkaline phosphatase activity, and osteoblast gene expression on the 7-day treated zirconia disks. RESULTS: Disks showed different surface morphologies after soaking for different time periods. As the SBF soaking time increased, the amount of HA coverage increased gradually, uniformly covering the disks by day 7. There was no difference in surface roughness between the two groups (P > .05). Cell proliferation was higher on the SBF-treated disks (P < .05). At 9 days, alkaline phosphatase activity was higher on the SBF-treated disks (P < .05). Alkaline phosphatase staining was significant on the SBF-treated disks (P < .05). A gene study revealed that alkaline phosphatase and osteocalcin showed higher expression in SBF-treated disks (P < .05); however, collagen type I and runt-related transcription factor 2 did not show significant differences between the two groups (P > .05). CONCLUSION: This study demonstrated that biomimetic deposition has an effect on the formation of HA on zirconia disks. The cell attachment, proliferation, and differentiation of SBF-treated zirconia disks was superior to that of nontreated disks, which indicates that SBF-treated zirconia implants have long-term clinical value.

Induction of tolerance against the arthritogenic antigen with type-II collagen peptide-linked soluble MHC class II molecules.

In murine collagen-induced arthritis (CIA), self-reactive T cells can recognize peptide antigens derived from type-II collagen (CII). Activation of T cells is an important mediator of autoimmune diseases. Thus, T cells have become a focal point of study to treat autoimmune diseases. In this study, we evaluated the efficacy of recombinant MHC class II molecules in the regulation of antigen-specific T cells by using a self peptide derived from CII (CII260-274; IAGFKGEQGPKGEPG) linked to mouse I-A(q) in a murine CIA model. We found that recombinant I-A(q)/CII260-274 molecules could be recognized by CII-specific T cells and inhibit the same T cells in vitro. Furthermore, the development of CIA in mice was successfully prevented by in vivo injection of recombinant I-A(q)/CII260-274 molecules. Thus, treatment with recombinant soluble MHC class II molecules in complex with an immunodominant self-peptide might offer a potential therapeutic for chronic inflammation in autoimmune disease such as rheumatoid arthritis. [BMB Reports 2016; 49(6): 331-336].

Eligibility and preference of new oral anticoagulants in patients with atrial fibrillation: comparison between patients with versus without stroke.

BACKGROUND AND PURPOSE: Recent randomized clinical trials (RCTs) have evaluated the benefit of new oral anticoagulants in reducing the risk of vascular events and bleeding complications in patients with atrial fibrillation (AF). However, abundant and strict enrollment criteria may limit the validity and applicability of results of RCTs to clinical practice. We estimated the eligibility for participation in RCTs of an unselected group of patients with AF. In addition, we compared features favoring new oral anticoagulant use between patients with versus without stroke. Randomized Evaluation of Long-Term Anticoagulation Therapy METHODS: We applied enrollment criteria of 4 RCTs (RE-LY, ROCKET-AF, ARISTOTLE, and ENGAGE-AF-TIMI 48) to 695 patients with AF taking warfarin, prospectively and consecutively collected at a university medical center; 500 patients with and 195 patients without stroke. Time in therapeutic range and bleeding risk scheme (anticoagulation and risk factors in atrial fibrillation) were also measured. RESULTS: The proportions of patients fulfilling the trial enrollment criteria varied, ranging from 39% to 72.8%, depending on the differences in indications/contraindications among studies and presence/absence of stroke. The main reasons for ineligibility for RCTs were hemorrhagic risk (anticoagulation and risk factors in atrial fibrillation [ATRIA] score) (10.8%-40.5%) and planned cardioversion (5.1%-7.7%) for nonstroke patients, and a low creatinine clearance (5.6%-9.2%) and higher risk of bleeding (15.2%-20.8%) for patients with stroke. When compared with nonstroke patients, patients with stroke showed a lower time in therapeutic range (54.4±42.8% versus 65.4±34.9%, especially with severe disability) and a high hemorrhagic risk (ATRIA score) (3.06±2.30 versus 2.18±2.16) (P<0.05 in both cases). CONCLUSIONS: Patients enrolled in RCTs are partly representative of patients with AF in clinical practice. When time in therapeutic range and bleeding tendency with warfarin use were considered, the use of new oral anticoagulants was preferred in patients with stroke than in nonstroke patients, but they were more likely to be excluded in RCTs.

Lack of association between hepatitis B virus pre-S mutations and recurrence after surgical resection in hepatocellular carcinoma.

Pre-S mutation of hepatitis B virus (HBV) is known to be a risk factor for hepatocarcinogenesis. A previous study suggested that pre-S mutation(s) may associate with increased recurrence after surgical resection. In the present study, 64 patients with HBV-related hepatocellular carcinoma (HCC) were categorized into two groups according to the presence or absence of pre-S mutation(s). The clinicopathological variables of the two groups were analyzed to assess the relationship between pre-S mutations and postoperative recurrence. Nineteen patients (29.7%) had pre-S mutations;13 had a pre-S deletion, three had a pre-S2 start codon mutation, two patients had both a pre-S deletion, and a pre-S2 start codon mutation, and one patient had a pre-S2 insertion. The two groups did not differ in terms of baseline clinicopathological parameters. Cirrhosis and satellite lesion(s) were predictive factors for postoperative recurrence and poor overall survival. Recurrence-free survival (P = 0.320) and overall survival (P = 0.238) did not differ significantly when pre-S mutations were present. In conclusion, this study did not find evidence supporting the notion that pre-S mutation(s) are associated with postoperative recurrence after surgical resection.

Effects of dietary supplementation with antimicrobial peptide-P5 on growth performance, apparent total tract digestibility, faecal and intestinal microflora and intestinal morphology of weanling pigs.

BACKGROUND: The increase in drug-resistant bacteria and the ban on antibiotic growth promoters worldwide make the search for novel means of preventing bacterial infection and promoting growth performance imperative. In this sense, antimicrobial peptides are thought to be ideal candidates owing to their antimicrobial properties, broad spectrum of activity and low propensity for development of bacterial resistance. The aim of the present study was to investigate the effect of dietary supplementation with antimicrobial peptide-P5 (AMP-P5) on weanling pig nutrition. RESULTS: A total of 240 weanling pigs were allotted to four treatments on the basis of initial body weight. There were four replicates in each treatment, with 15 pigs per replicate. Dietary treatments were negative control (NC, basal diet without antimicrobial), positive control (PC, basal diet + 1.5 g kg(-1) apramycin), basal diet with 40 mg kg(-1) AMP-P5 (P5-40) and basal diet with 60 mg kg(-1) AMP-P5 (P5-60). Pigs fed the PC or P5-60 diet showed improved (P < 0.05) overall growth performance, apparent total tract digestibility of dry matter, crude protein and gross energy and reduced (P < 0.05) faecal and intestinal coliforms compared with pigs fed the NC diet. CONCLUSION: The results obtained in this study indicate that dietary supplementation with 60 mg kg(-1) AMP-P5 has the potential to improve the growth performance and apparent total tract digestibility of nutrients and reduce coliforms in weanling pigs.

Mutation of a positively charged cytoplasmic motif within CD1d results in multiple defects in antigen presentation to NKT cells.

CD1d is an MHC class I-like molecule that presents glycolipid Ags to types I and II NKT cells. The YxxI motif in the cytoplasmic tail of CD1d contributes to its intracellular localization to the endolysosomal compartment and is important for Ag presentation to type I NKT cells. In this study, we identified the (327-329)RRR motif in CD1d and showed that it is critical for the control of CD1d intracellular trafficking and Ag presentation. The replacement of the arginines in this motif with alanines resulted in the extensive accumulation of CD1d in lysosomes but did not affect the cell surface expression. The defect in its cellular localization was accompanied by defects in Ag presentation to both type I and type II NKT cells. These results demonstrated that the (327-329)RRR motif of CD1d is required for proper cellular distribution of CD1d and optimal Ag presentation to both type I and type II NKT cells.

Protective potential of resveratrol against oxidative stress and apoptosis in Batten disease lymphoblast cells.

Batten disease (BD) is the most common form of a group of disorders called neuronal ceroid lipofuscinosis, which are caused by a CLN3 gene mutation. A variety of pathogenic lysosomal storage disorder mechanisms have been suggested such as oxidative stress, endoplasmic reticulum (ER) stress, and altered protein trafficking. Resveratrol, a stilbenoid found in red grape skin, is a potent antioxidant chemical. Recent studies have suggested that resveratrol may have a curative effect in many neurodegenerative diseases. Therefore, we investigated the activities of resveratrol at the levels of oxidative and ER stress and apoptosis factors using normal and BD lymphoblast cells. We report that the BD lymphoblast cells contained low-levels of superoxide dismutase-1 (SOD-1) due to the long-term stress of reactive oxygen species. However, when we treated the cells with resveratrol, SOD-1 increased to levels observed in normal cells. Furthermore, we investigated the expression of glucose-regulated protein 78 as an ER stress marker. BD cells underwent ER stress, but resveratrol treatment resolved the ER stress in a dose-dependent manner. We further demonstrated that the levels of apoptosis markers such as apoptosis induce factor, cytochrome c, and cleavage of poly (ADP)-ribose polymerase decreased following resveratrol treatment. Thus, we propose that resveratrol may have beneficial effects in patients with BD.

Could HBx protein expression affect signal pathway inhibition by gefitinib or selumetinib, a MEK inhibitor, in hepatocellular carcinoma cell lines?

Hepatitis B virus X (HBx) protein has been known to play an important role in development of hepatocellular carcinoma (HCC). The aim of this study is to find out whether HBx protein expression affects antiproliferative effect of an epidermal growth factor receptor-tyrosine kinase (EGFR-TK) inhibitor and a MEK inhibitor in HepG2 and Huh-7 cell lines. We established HepG2 and Huh-7 cells transfected stably with HBx gene. HBx protein expression increased pERK and pAkt expression as well as ß-catenin activity in both cells. Gefitinib (EGFR-TK inhibitor) inhibited pERK and pAkt expression and ß-catenin activity in both cells. Selumetinib (MEK inhibitor) reduced pERK level and ß-catenin activity but pAkt expression was rather elevated by selumetinib in these cells. Reduction of pERK levels was much stronger with selumetinib than gefitinib in both cells. The antiproliferative efficacy of selumetinib was more potent than that of gefitinib. However, the antiproliferative effect of gefitinib, as well as selumetinib, was not different between cell lines with or without HBx expression. Signal pathway activation by HBx might not be strong enough to attenuate the antiproliferative effect of EGFR-TK inhibitor. Future experiments are needed to understand the role of HBx protein expression in HCC treatment using molecular targeting agent.

Lactobacillus casei prevents impaired barrier function in intestinal epithelial cells.

The exact effect of probiotics on epithelial barrier function is not well understood. The aims of this study were to evaluate cytokine-induced epithelial barrier dysfunction in intestinal epithelial cells (IECs) and to study the role of probiotics in the prevention of epithelial barrier dysfunction. Caco-2 cells grown on transwell chambers were stimulated with tumor necrosis factor (TNF)-α or interferon (IFN)-γ basolaterally. Probiotic, Lactobacillus casei, was added 1 h before cytokine stimulation. MAPK inhibitors were added 15 min before L. casei stimulation. The electrical resistance and paracellular permeability of Caco-2 monolayers were measured. Distribution of zonula occludens (ZO)-1 protein was assessed by immunofluorescence, and Western blot analyses for ZO-1, p-Akt, and toll-like receptor (TLR) 2 were performed. Both TNF-α and IFN-γ stimulation on Caco-2 cells decreased transepithelial resistance (TER), increased epithelial permeability, and decreased ZO-1 expression of Caco-2 cells. In contrast, pretreatment of L. casei reversed the cytokine-induced dysfunction of TER, epithelial permeability, and ZO-1 expression. Reversal of cytokine-induced dysfunction of TER and intestinal permeability by L. casei was abrogated with MAPK inhibitor treatment. Lactobacillus casei stimulation on Caco-2 cells increased TLR2 and p-Akt expression. Probiotic, L. casei, prevents cytokine-induced epithelial barrier dysfunctions in IECs.