Clinical studies support the efficacy of programmed cell death 1 (PD-1) targeted therapy in a subset of patients with metastatic gastric cancer (mGC). With the goal of identifying determinants of response, we performed molecular characterization of tissues and circulating tumor DNA (ctDNA) from 61 patients with mGC who were treated with pembrolizumab as salvage treatment in a prospective phase 2 clinical trial. In patients with microsatellite instability-high and Epstein-Barr virus-positive tumors, which are mutually exclusive, dramatic responses to pembrolizumab were observed (overall response rate (ORR) 85.7% in microsatellite instability-high mGC and ORR 100% in Epstein-Barr virus-positive mGC). For the 55 patients for whom programmed death-ligand 1 (PD-L1) combined positive score positivity was available (combined positive score cut-off value ≥1%), ORR was significantly higher in PD-L1(+) gastric cancer when compared to PD-L1(-) tumors (50.0% versus 0.0%, P value <0.001). Changes in ctDNA levels at six weeks post-treatment predicted response and progression-free survival, and decreased ctDNA was associated with improved outcomes. Our findings provide insight into the molecular features associated with response to pembrolizumab in patients with mGC and provide biomarkers potentially relevant for the selection of patients who may derive greater benefit from PD-1 inhibition.

Salmonella gallinarum (SG) is a non-motile host-adapted salmonella that causes fowl typhoid, a severe systemic disease responsible for significant economic losses to the poultry industry worldwide. This study describes the application of a PCR-based signature-tagged mutagenesis system to identify in vivo-essential genes of SG. Ninety-six pools representing 1152 SG mutants were screened in a natural-host chicken infection model. Twenty presumptive attenuated mutants were identified and examined further. The identity of the disrupted gene in each mutant was determined by cloning of the DNA sequences adjacent to the transposon, followed by sequencing and comparison with the bacterial genome database. In vitro and in vivo competition indices were determined for each identified mutant and a total of 18 unique, attenuating gene disruptions were identified. These mutations represented six broad genomic classes: Salmonella pathogenicity island-1 (SPI-1), SPI-2, SPI-10, SPI-13, SPI-14 and non-SPI-encoded virulence genes. SPI-13 and SPI-14 are newly identified and designated in this study. Most of the genes identified in this study were not previously believed or known to play a role in the pathogenesis of SG infection in chickens. Each STM identified mutant showed competitiveness and/or virulence defects, confirmed by in vitro and in vivo assays, and challenge tests. This study should contribute to a better understanding of the pathogenic mechanisms involved in progression of disease caused by SG, and identification of novel live vaccine candidates and new potential antibiotic targets.

Preventing reactive gas species such as oxygen or water is important to ensure the stability and durability of organic electronics. Although inorganic materials have been predominantly employed as the protective layers, their poor mechanical property has hindered the practical application to flexible electronics. The densely packed hexagonal lattice of carbon atoms in graphene does not allow the transmission of small gas molecules. In addition, its outstanding mechanical flexibility and optical transmittance are expected to be useful to overcome the current mechanical limit of the inorganic materials. In this paper, we reported the measurement of the water vapor transmission rate (WVTR) through the 6-layer 10 × 10 cm(2) large-area graphene films synthesized by chemical vapor deposition (CVD). The WVTR was measured to be as low as 10(-4) g/m(2)·day initially, and stabilized at ∼0.48 g/m(2)·day, which corresponds to 7 times reduction in WVTR compared to bare polymer substrates. We also showed that the graphene-passivated organic field-effect transistors (OFETs) exhibited excellent environmental stability as well as a prolonged lifetime even after 500 bending cycles with strain of 2.3%. We expect that our results would be a good reference showing the graphene's potential as gas barriers for organic electronics.

In recent years, research on Brain-Computer Interface (BCI) technology for healthy users has attracted considerable interest, and BCI games are especially popular. This study reviews the current status of, and describes future directions, in the field of BCI games. To this end, we conducted a literature search and found that BCI control paradigms using electroencephalographic signals (motor imagery, P300, steady state visual evoked potential and passive approach reading mental state) have been the primary focus of research. We also conducted a survey of nearly three hundred participants that included researchers, game developers and users around the world. From this survey, we found that all three groups (researchers, developers and users) agreed on the significant influence and applicability of BCI and BCI games, and they all selected prostheses, rehabilitation and games as the most promising BCI applications. User and developer groups tended to give low priority to passive BCI and the whole head sensor array. Developers gave higher priorities to “the easiness of playing” and the “development platform” as important elements for BCI games and the market. Based on our assessment, we discuss the critical point at which BCI games will be able to progress from their current stage to widespread marketing to consumers. In conclusion, we propose three critical elements important for expansion of the BCI game market: standards, gameplay and appropriate integration.

Lipocalin-2 (Lcn2) is preferentially expressed in hepatocellular carcinoma (HCC). However, the functional role of Lcn2 in HCC progression is still poorly understood, particularly with respect to its involvement in invasion and metastasis. The purpose of this study was to investigate whether Lcn2 is associated with the epithelial-mesenchymal transition (EMT) in HCC and to elucidate the underlying signaling pathway(s). Lcn2 was preferentially expressed in well-differentiated HCC versus liver cirrhosis tissues, and its expression was positively correlated with the stage of HCC. The characteristics of EMT were reversed by adenoviral transduction of Lcn2 into SH-J1 cells, including the down-regulation of N-cadherin, vimentin, alpha-smooth muscle actin, and fibronectin, and the concomitant up-regulation of CK8, CK18, and desmoplakin I/II. Knockdown of Lcn2 by short hairpin RNA (shRNA) in HKK-2 cells expressing high levels of Lcn2 was associated with EMT. Epidermal growth factor (EGF) or transforming growth factor beta1 (TGF-b1) treatment resulted in down-regulation of Lcn2, accompanied by an increase in Twist1 expression and EMT in HCC cells. Stable Lcn2 expression in SH-J1 cells reduced Twist1 expression, inhibited cell proliferation and invasion in vitro, and suppressed tumor growth and metastasis in a mouse model. Furthermore, EGF or TGFb1 treatment barely changed EMT marker expression in SH-J1 cells ectopically expressing Lcn2. Ectopic expression of Twist1 induced EMT marker expression even in cells expressing Lcn2, indicating that Lcn2 functions downstream of growth factors and upstream of Twist1. Conclusion: Together, our findings indicate that Lcn2 can negatively modulate the EMT in HCC cells through an EGF (or TGF-b1)/Lcn2/Twist1 pathway. Thus, Lcn2 may be a candidate metastasis suppressor and a potential therapeutic target in HCC. (HEPATOLOGY 2013;58:1349-1361 L ipocalin-2 (Lcn2), also known as NGAL, belongs to the lipocalin protein family and was first purified from human neutrophils because of its association with gelatinase. 1 Lcn2 can exist as a 25-kDa monomer, 46-kDa disulfide-linked homodimer, and/or 135-kDa disulfide-linked heterodimer with neutrophil gelatinase. 2 Elevated Lcn2 expression has been observed in multiple human cancers including

Purpose: The poor survival rate of hepatocellular carcinoma (HCC) is in part due to the inability to diagnose patients at an early stage. Therefore, the aim of this study was to search for candidate serum marker for HCC and to test their ability to distinguish a HCC from benign liver disease. Experimental Design: Genome-wide analysis by a microarray in 40 HCC patients was done between HCC and paired nontumor liver tissues. Expression of cystatin B (CSTB) was examined by mRNA expression analysis and immunohistochemistry.The serum CSTB levels were measured using a sandwich ELISA method in four groups, including normal healthy subjects (group 1, n = 52) and patients with noncirrhotic chronic hepatitis (group 2, n = 53), cirrhosis (group 3, n = 43), and HCC (group 4, n = 62). Results: Microarray and statistical analyses identified 248 genes that were expressed differently between HCC and nontumor liver tissues. One of them, CSTB, was expressed preferentially in the HCCs compared with the nontumor tissues, 36 of 45 specimens (80%) by Northern blot and semiquantitative reverse transcription-PCR analyses. The serum CSTB level was much higher in HCC patients than in those with nonmalignant chronic liver disease (groups 2 and 3; P < 0.0001).The receiver operating characteristic curve indicated 5.34 ng/mL to be the optimal value for CSTB, and the sensitivity and specificity for this CSTB value were 85.5% (95% confidence interval, 74.2-93.1%) and 53.1% (95% confidence interval, 42.7-63.4%), respectively, in distinguishing between patients with HCC and those with nonmalignant chronic liver disease. Conclusion: CSTB is specifically overexpressed in most HCCs and is also elevated in the serum of a large proportion of HCC patients. CSTB or the combination of CSTB and a-fetoprotein may be a useful marker for diagnosing patients with HCC with a high sensitivity.

Novel biomarkers are needed for early detection and progression evaluation of hepatocellular carcinoma (HCC). The purpose of this study was to identify useful biomolecular markers for HCC. The 26 genes that encode membrane or secretory proteins were identified from cDNA microarray data. We further examined the expression of EFNA1 and its receptor EphA2 and determined their biological implications during the development and progression of HCC. The EFNA1 mRNA was overexpressed in most HCCs as compared with its expression in corresponding nontumor tissues (36 out of 40 cases, 90%), but EphA2 expression was noted in only half of the HCC tissues (20 of 40 cases, 50%). In most of the hepatoma cell lines, the EFNA1 protein expression was positively associated with alpha-fetoprotien (AFP) expression but inversely associated with EphA2 expression. Furthermore, EFNA1 levels were detectable in the supernatant of the cultured hepatoma cells and in the serum of patients with HCC. In contrast, EphA2 expression was prominent in highly invasive hepatoma cells, and its overexpression was significantly correlated with decreased differentiation (r 5 0.0248, p < 0.010) and poor survival (p 5 0.0453) for HCC patients. EFNA1 and EphA2 may be useful serum markers for the detection of HCC development and progression, respectively.Hepatocellular carcinoma (HCC) is the fifth most common cancer worldwide and it is known to be the third most common cause of cancer-related mortality. 1 The incidence of HCC is increasing in Western countries such as Europe and the Unites States. 2 One of the reasons for the high mortality is that HCC tumors are commonly detected at a stage when curative resection is no longer feasible because of intrahepatic and extrahepatic metastasis. The diagnosis of HCC currently relies on observation of a liver mass in radiology imaging studies such as ultrasonography, computed tomography scanning or magnetic resonance imaging. However, the diagnosis of small lesions is relatively inaccurate. 3 The only approach to screen for the presence of HCC in a high risk-population is the combination of serum alpha-fetoprotein (AFP) level determination and ultrasonography. 4,5 However, the AFP level test has low sensitivity and specificity, particularly in patients with smaller HCCs. Several biomarkers such as Desgamma carboxyprothrombin (DCP), lens cularis agglutininreactive AFP and glypican-3 (GPC3) have yet to be validated for their abilities to detect an early HCC. 6 Therefore, there is an urgent need to identify adequate biochemical markers for early detection and to evaluate the progression of HCC. Genome-wide microarray analysis offers a systemic approach for obtaining comprehensive information on the transcriptional profiles of HCC. Using microarray technology, we previously determined the molecular nature of multistep hepatocarcinogenesis and the specific genetic changes associated with the oncogenic differentiation of HCC. 7 In our previous study, we used a modified analytical approach to identify several molecular marke...

Oxidative stress is the main cause of cardiac injury during ischemia/reperfusion but the molecular mechanism for this process is unclear. In this study, it was found that hypoxia induces apoptosis in rat embryonic heart-derived H9c2 cells leading to the induction of GADD153, which is an apoptosis-related gene. Therefore, this study addressed the molecular role of GADD153 in hypoxia-induced apoptosis. The stable or inducible overexpression of GADD153 sensitized the H9c2 cells to apoptotic cell death. The results suggest that the transactivation domain of the GADD153 might be responsible for this cell execution and play a role in the nucleoplasmic localization of GADD153. The cells transiently transfected with the antisense GADD153 were more resistant to hypoxia-induced apoptosis than the vector control cells. Furthermore, GADD153 transcriptionally down-regulated the expression of the cardiac ankyrin repeat protein gene (CARP), which is a nuclear transcriptional co-factor that negatively regulates the expression of the cardiac gene. The ectopic expression of CARP in H9c2 cells increased the resistance to hypoxia-induced apoptosis. These results suggest that GADD153 overexpression and the concomitant down-regulation of CARP might have a causative role in the apoptotic cell injury of hypoxic H9c2 cells.