pylori as a class I human carcinogen, it now is well accepted that gastric infection see more by H. pylori is a risk factor for development of gastric cancer [8]. Although the precise pathogenetic role of H. pylori in gastric carcinogenesis remains unclear, it has been clarified that this organism contributes to modifications in epithelial cell proliferation, which may be the initiating event in a cascade culminating in the development of gastric cancer [9], but

it is not known whether the increased risk is due to the presence of mutant p53 generated by chronic gastritis or to a direct action of the bacteria on the p53 oncogene [10, 11]. The p53 gene mutation is associated with approximately 70% of tumors of different orignis [12, 13]. p53 gene serves as a “”gatekeeper of the cell”", for its role in preventing the accumulation of genetic alterations Osimertinib cost through the regulation of critical checkpoints between the end of G1 and the beginning of S to redirect cells with a mutation in the genome toward apoptosis or programmed cell death. This key oncogene thus prevents the perpetuation of a defective genome Mdivi1 datasheet and the development of a cancer [14]. Several recent studies have been published on the presence of p53 in patients with H. pylori infection, stomach cancer, or both. However, the conclusions are contradictory, and it has

been difficult to develop a single coherent hypothesis that can account for all findings communicated to date [15]. Palli et al [16] found p53 mutants in 33 of Thalidomide 105 cases of gastric cancer and Domek et al [17] worked with the hypothesis that tumorigenesis involves

deregulation of cell proliferation and apoptosis. These researchers investigated cell proliferation and apoptosis in the gastric epithelium of children infected with H. pylori, and found that the apoptotic index was significantly higher in patients with H. pylori gastritis than in patients with secondary gastritis or healthy control subjects. They also reported that apoptosis decreased when the bacterial infection was eradicated. Wu et al, reported the existence of different pathways of gastric carcinogenesis in different histological subtypes of gastric cancer and its progression, in which H. pylori infection can play an important role [18]. Hibi et al, concluded that persistent H. pylori infection caused gastritis, with degeneration and regeneration of the epithelium that increased cell proliferation and the accumulation of p53 [19]. This in turn increased instability of the gene, as reflected by the development of carcinoma, using immunohistochemical methods to investigate p53 expression, and concluded that expression is associated with histopathological phenotypes, and that genetic alterations may not be affected by H. pylori infection [20]. Chang et al, noted the possibility that H.

In order to maintain sustainable growth, a clean and renewable energy source is urgently required. Among all new types of energy sources, solar energy is the most promising one for it is safe, cheap, inexhaustible, and environment-friendly. In 1976, Carlson and Wronski [1] invented a new type of thin film solar cell that utilized amorphous silicon find more (a-Si) deposited from a glow discharge in silane (SiH4) and achieved a power conversion efficiency of 2.4% in AM-1 sunlight. After that, silicon thin film solar cells have been widely investigated in different ways and methods [2]. Compared with conventional solar cell, amorphous silicon thin film solar cell is low cost and could be deposited on various substrates

such as glass, stainless steel, ceramic plate, and plastic [3]. Studies focused on textured surface showed that it can buy Everolimus improve absorption

by reducing reflection. Textured surface can be conventionally obtained by either dry or wet ion etching [4–7]. In 2011, Wong and Yu [8] simulated a nanopillar-array-textured surface and came to a conclusion that it may enhance light absorption and increase the efficiency of the silicon-based solar cell. The effects of low-energy heavy ion irradiation on silicon thin film have been systematically studied during the past 50 years. During the irradiation, some traditional defects were generated; however, latent tracks, amorphous transition, or other special effects were not observed [9, 10]. Enhanced light absorption was obtained in works on n-type crystal silicon irradiated by high-energy Xe ion [11], which provided a promising method for the modification of amorphous silicon thin film. In this research, we coated a polystyrene (PS) sphere monolayer on glass substrate and fabricated silicon thin film via magnetic sputtering with glancing angle deposition (GLAD) in order to achieve periodically aligned C1GALT1 silicon nanopillar (PASiNP) arrays. The influences of silicon nanopillar diameter and Xe ion irradiation on the light absorption of thin film were studied. The mechanism of ion irradiation was also discussed. We replicate this nanostructure

by magnetic sputtering deposition with its advantage of controllable fabrication, and an expected selleck chemical enhancement in light absorption was observed. Methods Glasses were first cut into squares of about 3 × 3 cm2 in size and then thoroughly cleaned with acetone in an ultrasonic bath for 20 min. After washing off the residual acetone by deionized water, they were cleaned with ethanol in an ultrasonic bath for another 20 min. The glasses were immersed in H2SO4-H2O2 solution (3:1, v/v) for 8 h and then cleaned with deionized water in an ultrasonic bath for 30 min and with NH3-H2O2-H2O solution (1:1:3, v/v) for another 30 min. After that, glasses with hydrophilic surfaces were obtained [12]. PS nanospheres with different diameters of 200, 500, and 1,000 nm were selected here.

Angiogenesis is essential for progression,

invasion and metastasis of SCLC[11]. As a specific target of most tumors VEGF is a target gene of HIF-1 alpha and plays a main role in control of angiogenesis both in physiological and pathological situations, including tumor development and progression. It is mitogenic and angiogenic for endothelial cells, and it can also increase vascular permeability [12]. Identical with previous study [13] our study also found that VEGF-A was upregulated by HIF-1 alpha more than 6-fold in SCLC. But besides VEGF-A, there are several other genes associated with angiogenesis such as PDGFC, PLA2G4A, HMOX1, HMGA2 were upregulated by HIF-1 alpha. These genes were not reported in others literatures and therefore we think the upregulation of these genes may be specific to the angiogenesis Ralimetinib solubility dmso of SCLC when responding to HIF-1 alpha or hypoxia. Some genes had been reported to be found with differential expression in SCLC through microarray analysis. Amplification and overexpression of the MYC family of oncogenes such as MYC (c-Myc), H 89 clinical trial MYCN (N-Myc) and MYCL1 (L-Myc) occured in SCLCs [14] and was common in chemo-refractory disease[15]. In our study not MYC family but SLC family such as SLC6A2 and SLC9A2 were upregulated by HIF-1 alpha. Some genes as TAF5L, TFCP2L4, PHF20, LMO4, TCF20 and RFX2 that were

known to have transcription factor activities express highly in SCLC[16] but the genes that were upregulated by HIF-1 alpha are TRIM22, IRF9, MYOCD, ZNF277 and CREM from our study. Previous study also reported that the high expression of BAI3, D4S234E, DCX, DPYSL5 and GKAP1 which were PLX3397 related

to signal transduction were found in SCLC [16, 17]. In our study signal transduction factor IRS4 and GPER1 were upregulated by HIF-1 alpha more than 6.0-fold. As for IRS4 some researchers Oxymatrine have found that it plays an important role in proliferation/differentiation of tumors and exerts its actions through ERK and p70S6K activation in a ras/raf/MEK1/2 and PI3K/Akt independent manner and in a PKC-dependent way [18]. The GPER1 gene (also known as GPR30) represents an alternative estrogen-responsive receptor, which is highly expressed in tumors where estrogen and progesterone receptors are downregulated and in high-risk tumor patients with lower survival rates[19]. GPER1 is also an important mediator of some single transduction pathways contributing to promote proliferation, metastasis and aggressive behaviors of tumors that are induced by endogenous estrogens, including drugs like hydroxytamoxifen and atrazine or the environmental pollutant cadmium [20–22]. A novel finding different from previous study is that some genes encoding inflammatory response cytokines were upregulated. This maybe provides a broad molecular-biological basis for the inflammatory effect of SCLC.

Outside air temperature, humidity, and weather were recorded every 15 min during the time-trials using a WS9623 Wireless 868 MHz Weather Station (La

Crosse Technology, France). Data analyses Performance was assessed via overall time to selleckchem complete the time-trial. The cyclists’ uphill time splits were also used as a measure of performance to account for any variation in skill in descending the hills. Plasma [Na+] (mmol.L-1), haematocrit, and blood glucose values (mmol.L-1) were analysed via the i-STAT point of care analyser (Abbott Point of Care Inc, Illinois, USA) and recorded in the field. Sweat sodium and chloride concentration (sweat [Na+], sweat [Cl-]) was analysed in small batches through a Cobas C311 module (Roche Diagnostics, Basel, Switzerland) using the Ion Selective Electrode (ISE) DMXAA solubility dmso technique (mean CV = 2.01 ± 1.59%). Sweat sodium concentrations were then extrapolated to whole body sweat sodium SRT1720 price losses using the calculations of Patterson et al. [17]. To ensure contamination of the patches nor leaching from the skin had not occurred sweat potassium was measured and all samples were within the

normal range [18]. Urine osmolality was measured via freezing point depression (Osomat 030, Genotec GmbH, Baden-Wurttemberg, Germany), to indicate hydration status. Subjective feelings of thirst were indicated on a 100 mm visual analogue scale, which was used as a rating from 0 (not thirsty at all) to 100 (extremely thirsty) [15]. Statistical analysis Statistical analyses were performed using Stata Version 11.2 (StataCorp, Texas, USA). Normality of the data was evaluated using a Shapiro-Wilks test, and difference in variance was assessed by two-group variance comparison tests before all comparisons. Multivariate regression was used to assess the effect of sodium

supplements on exercise performance and plasma [Na+]. Differences in overall time and uphill time were compared whilst controlling for temperature and weather (wet or dry road). The difference in absolute (mmol.L-1) Thalidomide and relative (%) plasma [Na+] change was analysed controlling for average heart-rate. A paired t-test was also used to investigate differences in plasma [Na+] from pre-race to post-race within each intervention. Urine and sweat concentrations were well distributed and the absolute (mmol.L-1) and relative (%) change in electrolytes in each were analysed using a Student’s t-test. Changes in body mass, haematocrit, plasma volume change and fluid intake were assessed using multivariate regression controlling for mean heart rate and temperature. Statistical significance was set at p ≤ 0.05. If a relationship was close to statistical significance, a Cohen’s d effect size was also calculated. Data is reported as mean ± standard deviation (SD). Results Descriptive characteristics of the participants are shown in Table 1. Participants were lean, with a mean sum of eight skinfolds of 82.

Tplain was positioned in the top #selleck inhibitor randurls[1|1|,|CHEM1|]# left quadrant. Figure 3 PCA plot showing the clustering of the samples. The figure shows a PCA plot based on taxonomic (phylum level) and metabolic (SEED subsystems, level I) parameters combined. The geochemical

[25] parameters were overlain using the envfit function of the vegan library in R. The first principal components accounted for 95 % of the variation in the dataset, while the second principal component accounted for 3 %. All metagenome data were given as percent of total reads. The geochemical parameters were normalized by dividing with the standard deviation and subtracting the smallest number from all numbers in each row. Plot A: the metagenomic parameters are represented by red arrows. Labels are shown for parameters with Euclidian distance over 0.1 from origin. The geochemical parameters are represented by blue arrows. Only the most significant geochemical parameters are shown (p-value < 0.1). Plot B: is an excerpt of plot A, magnifying the central region of the plot. Labels for all metagenomic parameters with Euclidian distance over 0.02 are included. The first principal component (PC1) accounted for 95% of the variance in the dataset. Along the PC1 axis Tpm2 was the Troll sample most similar to the Oslofjord samples, while Tplain and Tpm1-2 were positioned furthest away. Tpm3 and Tpm1-1 were placed at an intermediate position. The

abundance of Proteobacteria was the most important parameter for the positioning of sites along PC1. Proteobacteria, as well as Thaumarchaeota, Planctomycetes Bromosporine solubility dmso and Actinobacteria had high negative scores along this axis. The analysis thereby indicated relatively high abundances of these taxa at the sites placed on the left side of the plot, especially Tpm1-2 and Tplain (Figure 3, Additional file 5: Table S3). Firmicutes, Euryarchaeota, Chloroflexi and Viruses all had high positive scores along PC1 indicating that the samples placed in the right section of the PCA plot (OF1, OF2 and Tpm2) had relatively high abundances of these taxa compared to the other sites. Although

Tpm2 grouped with the Oslofjord Rucaparib in vitro samples along PC1, it was separated from the Oslofjord samples by PC2. While Chloroflexi, Euryarchaeota, Thaumarchaeota and Firmicutes had high negative scores along PC2, Bacteroidetes, Actinobacteria and Planctomycetes had the highest positive scores along this axis and can therefore be considered as important parameters for the placement of the Oslofjord samples and Tplain in the top half of the plot. Concerning the carbon sources, the geochemical parameters supported a positive correlation between hydrocarbons (< n-C32) and the Troll samples, while concentrations of bicarbonate and TOC were positively correlated with the Oslofjord samples (Figure 3, Additional file 4: Table S2 and Additional file 6: Figure S3).

jejuni 81-176 sequences; restriction recognition sites introduced for cloning purposes are underlined, complementary fragments of primers Cjj46mwR and Cjj43mwL are marked with italics. Point mutated VE821 nucleotides in primers are marked with small letters. Orientation of the primers

Ulixertinib (Fwd states for forward/Rev – for reverse) refers to the orientation of particular C. jejuni gene studied. RT-Cj primer was designed on the basis of C. coli 72Dz/92 dsbI nucleotide sequence (there are 2 nucleotide changes compared to the nucleotide sequence of its orthologue from C. jejuni 81-176). All vectors containing transcriptional fusions of putative dsb gene promoter regions

with a promotorless lacZ gene were constructed using the pMW10 E. coli/C. jejuni shuttle vector. DNA fragments were amplified selleck products from C. jejuni 81-176 chromosomal DNA with appropriate pairs of primers (listed in Table 2). Next, PCR products were cloned in the pGEM-T Easy vector (Promega), excised by restriction enzymes and subsequently cloned into pMW10, forming transcriptional fusions with the downstream promoterless lacZ reporter gene. Correct construction of the resulting shuttle plasmids was confirmed by restriction analysis and sequencing. Morin Hydrate All recombinant

plasmids, as well as the empty pMW10, were introduced into C. jejuni 480 cells by electroporation. Construction of a pUWM1072 plasmid containing dsbI without dba under its native promoter was achieved by PCR-amplification of the 520 bp chromosomal DNA fragments containing the dba-dsbI promoter sequences (primer pair Cj19LX-2 – Cj18Nde-Rev) and cloning it into pBluescript II SK (Stratagene), using XbaI/PstI restriction enzymes. Subsequently the dsbI coding sequence (1792 bp) was PCR-amplified using the Cj17Nde – Cj16RS primer pair, cloned into pGEM-T Easy (Promega) and finally, using NdeI/SalI restriction enzymes, transferred into pUWM1072 in the native orientation, generating the plasmid pUWM1100. The whole insert (2316 bp) was then cloned into a shuttle E. coli/C. jejuni vector pRY107 [27] using SalI/XbaI restriction enzymes. The resulting, plasmid pUWM1103, whose correct construction was verified by sequencing, was used for complementation assays in C. jejuni Δdba-dsbI::cat mutant cells. Point mutations were generated using a Quick-Change site-directed mutagenesis kit, following the supplier’s recommendations (Stratagene).

In this study we demonstrate that inclusion migration along microtubules promotes inclusion fusion. Interestingly, although this dynein dependent migration was required for the normal timing of inclusion fusion, inhibition

of this trafficking was eventually overcome later during infection. Methods Organisms and cell culture All cells were obtained from the American www.selleckchem.com/products/pu-h71.html Type Culture Collection. Cell lines are: McCoy (McCoy B, CRL-1696), HeLa (HeLa 229, CCL-2.1), Cos7 (COS-7, CRL-1651) and neuroblastoma (N1E-115, CRL-2263). Chlamydia trachomatis serovars are: L2 (LGV 434), G (UW-524/CX) and J (UW-36/CX). C. trachomatis were propagated in McCoy or HeLa cells. EBs were purified by Renografin (Bristol-Myers Squibb, New York, NY, USA) density gradient centrifugation as previously described [10, 11]. HeLa and Cos7 cells were grown in RPMI-1640 (Lonza, Basel, Switzerland) supplemented with 10% FBS (Gibco/Life Technologies, Grand Island, NY, USA) and 10 μg/mL gentamicin (Gibco). McCoy and neuroblastoma cells were grown in DMEM (Lonza) supplemented with 10% FBS (Gibco) and 10 μg/mL gentamicin (Gibco). All cells were grown in 5% CO2 at 37°C. Infections All infections were carried out as follows unless otherwise noted. Cells were incubated with C. trachomatis EBs in Hank’s selleck balanced salt solution (HBSS) (FK866 Invitrogen/Life Technologies,

Grand Island, NY, USA) for 30 min at 22°C. The inoculum was replaced with prewarmed, 37°C, complete media. For nocodazole treated cells, the inoculum was replaced with prewarmed, 37°C, complete Rebamipide media containing 5 μg/mL nocodazole. Infected cells were incubated in 5% CO2 at 37°C. Synchronized infections Cells were incubated with C. trachomatis EBs in HBSS (Invitrogen) at MOI = 1000 for 5 min at 22°C. The cells were washed three times with HBSS plus 100 μg/mL heparin (Pharmacia, Peapack, NJ, USA) and twice with HBSS without heparin. Prewarmed, 37°C, complete media was added and infected cells

were incubated in 5% CO2 at 37°C. Transfections and plasmids HeLa cells were grown on 12 mm number 1.5 borosilicate glass coverslips coated with Poly-L-lysine (Sigma-Aldrich, St. Louis, MO, USA) to obtain a monolayer of approximately 65% confluency. Transfections were carried out using Lipofectamine 2000 (Invitrogen) according to the manufacturer’s instructions. Expression from the transfected vectors was allowed to proceed for at least 24 h prior to experimentation. Expression vectors used were pEGFP-C3 (Clontech, Mountain View, CA, USA), EB1-GFP and EB1.84-GFP. The EB1-GFP plasmid was a kind gift from Dr Jennifer S. Tirnauer, University of Connecticut Health Center. The EB1.84-GFP plasmid was generated by PCR cloning of the N terminal end of EB1 and cloning into pDest-NGFP as described by Askham et al. [12]. Micro-injections Cos7 cells were grown on 25 mm number 1.

In fact, the binding EGFR/ligand leads to activation of the TK, thus inducing cell growth, inhibition of apoptosis, angiogenesis, invasion and metastasis [2]. EGFR overexpression in non small cell lung cancer (NSCLC) and colorectal cancer (CRC) is a frequent event related to a poor outcome [3]. In the last few years, many

clinical trials have proven the efficacy of EGFR-targeted therapies in the management of several cancers, including breast, colon, GSK1210151A solubility dmso pancreas, head and neck, renal, and lung carcinomas. Multiple therapeutic strategies have been developed to target EGFR, including monoclonal antibodies (MoAbs), tyrosine kinase inhibitors (TKI), ligand-toxin conjugates, and antisense oligonucleotides. Cetuximab and panitumumab are two MoAbs which are active against the ligand selleck inhibitor binding site of EGFR with high specificity and higher affinity for EGFR than the natural ligands TGF-α and EGF, and are now considered

as one standard option for patients with advanced CRC in the first or second line of treatment [4, 5]. Indeed, the anti-EGFR click here erlotinib and gefitinib have undergone extensive clinical testing demonstrating clinical activity in NSCLC [6]. In this context, there is a need for methods enabling response prediction in order to select those patients most likely to benefit from treatment. Therefore, the diagnostic approach of pathologists is changing, leading to an integrated morphological and molecular diagnosis. EGFR overexpression does not seem a good predictor of response to Sucrase treatment both in NSCLC and CRC [7, 8], even though some controversial results are reported [9]. According to poor clinical information obtained from the immunohistochemistry (IHC), the interest in EGFR

gene status increased after Moroni et al [10] proposed that in CRC the response to anti EGFR treatment with cetuximab is related to EGFR gene copy number (GCN) and Lynch et al [11] showed that, in advanced NSCLC, in-frame deletion or missense mutations in the EGFR TK domain can predict the response to therapy with gefinitib. In addition, several authors [12, 13] reported that, in metastatic CRC (mCRC), an increased EGFR GCN or mutations of genes (i.e. k-ras) responsible for downstream signalling are important determinants of response or resistance to anti-EGFR antibodies, such as cetuximab and panitumumab. Specifically, cetuximab has proven efficacy in the treatment of mCRC, but also in NSCLC with squamous cell histology [14]. Although fluorescence in situ hybridization (FISH) is the “”gold standard”" method to detect EGFR gene amplification, this technique presents some disadvantages since the fluorescent signal is not stable and morphological features are difficult to visualize. In contrast, chromogenic in situ hybridization (CISH) utilizes a peroxidase reaction to detect the locus of interest and can be interpreted by standard light microscopy in the context of morphology [15].

neg C Z Z 15 Multinodular goiter N/C N/C F Z 16 Follicular adenoma C N F Z 17 Multinodular goiter N/C N/C F Z 18 Multinodular goiter N/C N/C F F 19 Papillary cancer & Hashimoto C C F Z C = cytoplasmic; N = nuclear; F = focal; Z = zonal; ND = not determined; neg. = Tozasertib manufacturer negative Biopsy tissues used for immunohistochemical analyses were obtained from normal tissue adjacent to diseased areas. Bucladesine samples were immediately

frozen in liquid Nitrogen and stored at -80°C. On the day of analysis, tissue samples were gradually set to the temperature of -30°C for cryostat procedure. Seven sections were cut from each sample. The immunoperoxidase method was applied with Vector reagents utilizing the following

primary antibodies: a) the anti-p53 polyclonal antibody CM-1 (Novocastra Laboratories Ltd) dilution 1:1000, b) the anti-STAT3 polyclonal antibody C-20 sc-482 clone (Santa Cruz Biotechnology) dilution 1:1000, c) the anti-CK19 monoclonal antibody b170 (Novocastra Laboratories Ltd) dilution Caspase Inhibitor VI ic50 1:100, d) the anti-gp130 polyclonal antibody H-255 (Santa Cruz Biotechnology) dilution 1:250. The staining pattern was evaluated in epithelial cells both in terms of percentage of stained cells and staining intensity. In terms of percentage of stained cells, samples were classified as diffuse, zonal, focal and negative when the % of positive cells was >50%, between 10-50%, <10% and 0%, respectively. In terms of staining intensity, samples were subdivided into three categories: 1 + (low), 2 + (intermediate)

and 3 + (high). Results The results of immunohistochemical analyses are shown in Table 1. Except for case number 8 (multinodular goiter) that was negative for both STAT3 and p53 expression, and case number 14 (papillary SPTBN5 carcinoma) which was negative for STAT3, a diffuse pattern with an intermediate intensity in both nuclear and/or cytoplasmic localizations was observed in all the samples analyzed. An exclusive cytoplasmic localization of STAT3 was seen in 7 cases while a nuclear/cytoplasmic staining was detected in 10 cases. As for p53, three cases displayed an exclusive nuclear staining, 8 cases showed an exclusive cytoplasmic localization, 7 cases showed a nuclear/cytoplasmic positivity [Figure 1] and one case displayed no staining. gp130 staining was negative in two cases (3 and 8) while a zonal or focal membrane and cytoplasmic staining distribution of intermediate intensity (2+) was observed in most of the cases [Figure 2]. Cases 7, 15 and 19 showed an intense (3+) staining. Cytokeratin 19 (CK19) could not be determined in case 3, while 7 samples were negative, 8 showed a focal and 3 a zonal cytoplasmic distribution of intermediate intensity (2+).

As shown in Figure 3A, 4D10 specifically reacted with the synthetic peptide PL10, whereas control antibody 4G2 (anti-flavivirus E mAb) did not reacted with PL10. For the sensitivity binding assay, the synthetic peptide PL10 bound the antibody in a concentration-dependent manner. Two control peptides PH10 (3LTTRGGEPHM12) and PM10 (SQNPPHRHQS) were not reactive

(Figure 3B). Figure 3 Properties analysis of synthetic peptide PL10. (A) Specific reactivity of PL10 with antibody 4D10 (anti-DENV1-4 prM mAb). The synthetic peptide PL10 could react with mAb 4D10 but control antibody 4G2 (anti-flavivirus E mAb) could not. (B) The sensitivity binding assay of synthetic peptides PL10 and two control peptides (PH10 and PM10) with mAb 4D10. The synthetic peptide PL10 bound the antibody in a concentration-dependent manner, but two control peptides had no reactivity with 4D10. (C) ELISA reactivities of synthetic peptide PL10 with immunized mice sera. Synthetic selleck inhibitor peptide PL10 was recognized by anti-DENV1-4 mice sera, whereas it was not recognized by anti-JEV mice sera and normal mice sera (NMS). (D) Competitive inhibition of phage clone binding to mAb 4D10 by synthetic peptide PL10. Competitive ELISA was performed using PL10 as competitor of its corresponding phage clones.

The percentage of inhibition is also shown. (E and F) ELISA reactivities click here of synthetic peptide PL10 with serum samples from 20 until DENV2-infected patients (E) and 20 BX-795 cost healthy adults (F). PH10 and PM10 were used as control. Data are expressed as means of at least three independent experiments. The error bars represent standard deviations (SD).

If there is no error bar, it is not that no variations among three independent experiments but that the variations are too small to show in the figure. * P < 0.05 vs PL10 at 0.1 μg. We next evaluated whether the synthetic peptide PL10 could be react with anti-DENV1-4 mice sera. Synthetic peptide PL10 was recognized by anti-DENV1-4 mice sera, whereas it was not recognized by anti-JEV mice sera and normal mice sera (NMS) (Figure 3C). We concluded that synthetic peptide PL10 is a DENV serocomplex cross-reactive epitope-based peptide. To confirm further the phage-displayed peptide was the epitope of antibody 4D10, a peptide competitive-inhibition assay was performed to determine whether the PL10 peptide competed with corresponding phage clones for reactivity with 4D10. The reaction activity of antibody 4D10 with the corresponding phage clones was inhibited markedly by PL10 at 0.1 μg per well with the inhibition percentage from 34% to 69% (Figure 3D). The results showed that the synthetic peptide and corresponding phage clones competed for the same antibody-binding site. Together, these findings suggest that 4D10 recognizes a new epitope on the N-terminal segment of DENV1-4 prM protein. Then, we evaluated the reactivity of synthetic peptide PL10 with DENV2 patient serum samples.