5 months (standard deviation 4.0 months). The time between the first and third QFT was, on average, 19.8 months (standard deviation 5.5 months). No association was observed between the time span of the QFTs and the probability of conversion or reversion in the QFT (data not shown). Nine HCWs were diagnosed with active TB, all but MRT67307 cost two were acid-fast bacillus (AFB)-positive, culturally confirmed cases. In one person, diagnosis was based solely on PCR (Table 6). All persons with active TB were positive in the first QFT. The TST was ≥15 mm in seven and 10–14 mm in two of them. Seven HCWs had

pulmonary TB, one pleural TB, and one skin TB. Six active TB cases were diagnosed within 2 months of the first QFT. The other three cases were diagnosed three, seven, and 19 months after the first positive QFT. In one case, a second QFT was performed at the time of diagnosis 3 months after the first QFT and an increase from 0.51 to 1.96 IU/mL was observed. The median of the INF-γ concentration in those with actual selleck chemicals llc pulmonary TB was 2.26 IU/mL, the minimum was 0.51 IU/mL, and the selleck chemicals maximum 6.32 UI/mL. For the HCW with pleural TB the INF-γ in the first QFT was 0.42 IU/mL and in the skin TB case it was >10 IU/mL. After diagnosis and treatment,

a reversion occurred in the patient with pleural TB and a sharp decline occurred in the HCW with cutaneous TB (>10–1.04 IU/mL). Baf-A1 order For the other six cases, increases and decreases of INF-γ concentration were observed three times, respectively. A positive QFT led to diagnosis in four HCWs with no symptoms. In the other 5 HCWs with pulmonary, active TB, typical symptoms such as cough, fever, weakness, or weight loss were observed along with a positive QFT. Table 6 Characteristics of the 9 HCWs diagnosed with active TB TB Gender Age TST mm 1st QFT IU/mL Months between 1st QFT and diagnosis 2nd QFT IU/mL Symptoms at first QFT Pneumal Female 26 17 0.51 3 1.96* None Pneumal Female 39 18 3.97 <1 6.29 None Pneumal Female 25 16 6.32 19 1.30

Cough Pneumal Female 29 17 2.11 <1 3.28 Cough Pneumal Female 25 13 1.30 <1 1.22 Cough, fever Pneumal Female 31 22 0.92 7 0.56 Cough, weakness, weight loss Pneumal Female 25 14 2.41 <2 3.57 None Pleurala Male 26 20 0.42 <1 0.10 None Cutaneousb Female 50 21 >10 <1 1.04 Skin lesion * In all but the first case, the second QFTs were performed after diagnosis a Positive PCR, if not indicated otherwise all cases were AFB-positive and culturally confirmed bCulturally confirmed Discussion Our study is the largest follow-up study for serial testing to date. Furthermore, it is also the only study on serial testing that actually observed active TB cases, thus allowing conclusions about test interpretation in serial testing to be based on these findings.

Several new chemotherapy agents are being tested in combination with radiation, but the best chemotherapy remains to be determined. The fate of irradiated cells is believed to be controlled by the network of signaling elements that lead to different modes of cell death or survival. Many stress-responsive genes are inducible by IR [18, 19]. These radiation-inducible genes are believed to have effects on the chemosensitivity Rabusertib ic50 of tumor cells [13, 20]. To determine the correlation between radio-resistance and sensitivity to chemotherapeutic drugs in esophageal cancer cells, we then analyzed the chemosensitivity of

EC109 and EC109/R cells with chemotherapeutic drugs cisplatin, 5-fluorouracil, doxorubicin, paclitaxel or etoposide. EC109/R, which survived 80 Gy irradiation, became more sensitive to different concentrations of 5-fluorouracil, doxorubicin, paclitaxel and etoposide, but maintained tolerance to cisplatin, as assessed by MTT assay (Figure 4). These findings suggest that cellular resistance to ionizing radiation have effects on the chemotherapeutic drug sensitivity in esophageal cancer cells. Several genes associated with cellular sensitivity to anticancer drugs have been selected for esophageal cancer. They were B4GALT5 (UDP-Gal: βGlcNAc β1,4-galactosyltransferase, polypeptide 5 gene), UGCG (UDP-glucose ceramide

glucosyltransferase gene), and XBP1 (X-box binding protein 1 gene) for 5-fluorouracil, selleck compound NRCAM (neuronal cell adhesion molecule gene) for doxorubicin, ARFRP1 (ADP-ribosylation factor related protein 1 gene), IFITM1 (interferon induced transmembrane protein 1 gene), KIAA0685, and SIPA1L2 (signalinduced proliferation-associated 1 like 2 gene) for cisplatin [14]. Fractionated irradiation might induce cellular sensitivity related gene and protein expression in human tumor cell lines. The fact

that drug Ceramide glucosyltransferase sensitivity is determined by multiple genes required a better understanding of the intricate network of the selected genes in the expression levels. Fractionated radiation treatment has also been reported to cause drug resistance in ovarian carcinoma cells [21] and ascites tumor cells [22]. It can induce functionally relevant ATR inhibitor multidrug resistance gene and protein expression in human tumor cell lines [13]. There are multiple factors that contribute to cisplatin resistance, but alterations of DNA repair processes have been known for some time to be important in mediating resistance [23, 24]. The most important DNA repair pathways involved in the cisplatin response are nucleotide excision repair (NER) and mismatch repair (MMR). MSI, which results from disorder of the MMR system and loss of MLH1 protein, is frequently induced during cisplatin-based chemotherapy [25]. Data have shown that suppression of ERCC1 expression enhances or restores cisplatin sensitivity, and combination of p53 inactivation and MMR deficiency results in cisplatin resistance [26].

Amplimers were column purified (Qiaquick PCR purification kit, Qiagen) and digested overnight with DpnI (Roche). Digests were pellet paint precipitated and the half of the digest directly electroporated into NZ9000. Between 200 and 1000 colonies were obtained per transformation. The protocol was repeated to combine SDM changes. From the final mutagenized plasmids, BglII/BstXI fragments containing the LRR region of InlA were excised and ligated into complementary digested pNZBinlA WT. Isolation of cell wall proteins Cell wall proteins were https://www.selleckchem.com/products/Vorinostat-saha.html isolated from nisin induced 10 ml NZ9000+pNZBinlA

WT culture as MX69 described by previously [22], except cells were rendered as protoplasts for 1 hr at 30°C without mutanolysin. Blotted proteins were probed with the InlA specific monoclonal antibody described by Hearty et al [23]. Random Bank of inlA mutants in NZ9000

The generation of a randomly mutated inlA 4SC-202 bank between amino acids 74 and 512 (containing the LRR) of InlA was accomplished by error prone PCR with Mutazyme II (Stratagene). Plasmid DNA (pNZBinlA WT) was used as template in the reaction (primers IM317 and IM318) and a 1.3 kb fragment amplified between two naturally occurring restriction sites (BglII and BstXI). From the mutagenesis reactions, four different mutation rates by varying the amount of template used ((iii) 1000 ng (iv) 250 ng (v) 10 ng and (vi) 0.1 ng). This equates to a sliding scale of increasing mutation frequency. Each amplimer pool was digested with BglII and BstXI and ligated into complementary digested pNZ8048binlA. The ligations (100 ng of pNZB with 240 ng of inlA) were pellet paint precipitated and electroporated into electrocompetent NZ9000 (repeated twice). For each pool a total of 40,000 colonies were obtained with plasmid religations accounting for 0.125% of the total (about 50 colonies per 40,000). The colonies from each mutation frequency were pooled and frozen at -80°C. From each mutation frequency, 10 individual colonies were subjected to plasmid isolation as described above and the mutated region sequenced to access the level of

mutagenesis. CT-26 and Caco-2 Inositol monophosphatase 1 invasion assays Overnight cultures of NZ9000 containing pNZB only or pNZBinlA derivatives (Figure 1a) were induced as described above. A one ml aliquot was then pelleted at 4,000 × g for 5 min and resuspended in 1 ml of DMEM. Cells were centrifuged, resuspended in fresh DMEM and then diluted to a multiplicity of infection of 25:1. L. monocytogenes cells were grown as described previously prior to invasion [20]. CT-26 [24] and Caco-2 cells were seeded at 2 × 104 and 1 × 105 cells, respectively and grown for 4 days until confluency in 24 well plates (Falcon). On the day prior to use, antibiotics were removed from the media. On the day of use, cells were washed twice with DMEM to remove FBS.

2). The tested genes showed the same trend in expression by Northern as

in the microarray. Figure 2 Northern blot analyses of Adriamycin in vitro CcpA-dependent genes. A, Transcription of genes showing differential expression in the ccpA mutant in the absence of glucose. Gene expression at an OD600 of 1 in strain Newman and its ΔccpA mutant is shown. B, Transcription of CcpA-dependent, glucose-dependent genes in strain Newman and its ΔccpA mutant. Cells were grown to an OD600 of 1, cultures where split and glucose added to one half (+), while the other half remained without glucose (-). RNA was sampled at an OD600 of 1, and after 30 min. RNA loading is represented by the intensity of the 16S rRNA. Data are representative for at least two independent experiments. MA, microarray data. CcpA-dependent AZD3965 nmr SC75741 chemical structure differential gene expression without glucose addition Genes showing an altered expression in the

ΔccpA mutant compared to the wild-type when growing in LB alone, without glucose addition, are listed in Additional files 1: Genes with lower expression in wild-type versus ΔccpA mutant, and 2: Genes with higher expression in wild-type versus ΔccpA mutant. These genes made up the largest regulatory group found in our study (226 genes). Only a minor part of this group of genes (38 out of 226) contained putative cre-sites in their promoter regions or were part of operons with putative cre-sites, suggesting that CcpA may affect the expression of the majority of these genes indirectly. Such indirect effects may reflect differences in the generation of metabolites due to ccpA inactivation, which might serve as cofactors for the regulation of further genes, and/or to a CcpA-dependent control of regulatory

proteins or RNAs. Our findings suggest that glucose-independent effects due to CcpA might play a particularly important role in S. aureus. For a better understanding, the genes of this category were grouped into functional for classes (Fig. 3A). While unknown proteins represented the largest group (61 genes), this group was followed by proteins of carbon metabolism (26 genes), transport/binding proteins and lipoproteins (25 genes), and proteins of amino acid metabolism (19 genes). Figure 3 Functional classes of CcpA-dependent genes. Functional classification according to the DOGAN website [26] of genes that were found to be regulated by CcpA in a glucose-independent (A) or a glucose-dependent way (B).

2008). Several studies correlated improved plant tolerance to abiotic stresses upon pathogenic or mutualistic microbial infections with an observed increase in antioxidant or osmolyte concentrations and/or in antioxidant enzymes activities (Rouhier and Jacquot 2008). This may Roscovitine in vivo explain the development of systemic acquired resistance in plants following pathogenic infections where healthy plant parts gain more resistance to a subsequent infection by either the same or another microbe (Singh et al. 2011). The root colonizing endophytic fungus Piriformospora indica was discovered in association with woody shrubs in the Indian Thar GS-9973 in vitro desert and was found to improve plant fitness of a variety

of host plants by growth enhancement under normal and stress conditions (Verma et al. 1998; Schäfer et al. 2007). The fungus was reported to activate nitrate reductase

MK0683 and glucan-water dikinase enzymes resulting in increased nitrate acquisition and/or starch degradation in Arabidopsis and tobacco roots (Sherameti et al. 2005). Further studies indicated involvement of cytokinins in P. indica induced growth promotion of Arabidopsis plants, while auxins had little or no effect (Vadassery et al. 2008). In addition to growth promotion, P. indica, originally isolated from desert plants, was found to induce drought stress tolerance of Arabidopsis and Chinese cabbage (Brassica rapa) by stimulation the expression of stress-related genes in leaves (Oelmüller et al. 2009; Sun et al. 2010). In Chinese cabbage colonized by P. indica the activities of peroxidases, catalases and superoxide dismutases in the leaves were increased within 24 h in response to drought cAMP stress. The fungus also increased the amount of chloroplast-localized Ca2+ sensing receptor protein, which regulates stomatal function in response to elevations of external Ca2+ by modulating

cytoplasmic Ca2+ concentration (Weinl et al. 2008; Sun et al. 2010). Furthermore, the drought induced decrease in photosynthetic efficiency and the degradation of chlorophylls and thylakoid proteins were delayed (Sun et al. 2010). P. indica also induced salt tolerance to a salt-sensitive barley cultivar (Hordeum vulgare) by increasing the rate of metabolic activity to compensate salt-induced inhibition of leaf metabolism (Criddle et al. 1989; Baltruschat et al. 2008), by induction of antioxidant enzymes (Baltruschat et al. 2008), and by enhancing the ratio of reduced to oxidized ascorbate (Waller et al. 2005). The latter neutralizes oxygen free radicals and acts as a primary substrate in the ascorbate-glutathione cycle to detoxify hydrogen peroxide (Foyer and Noctor 2000). It may also act by accelerating root elongation and increasing root biomass (Córdoba-Pedregosa et al. 2005). Furthermore, P. indica enhanced the biosynthesis of polyamines and lowered that of ethylene by increasing methionine synthase levels (Peškan-Berghöfer et al.

Dietary intake was not controlled but participant’s dietary intake was recorded prior to each testing session and analyzed for energy intake and macronutrient content. Participants were instructed to maintain their normal resistance-training program and maintain training logs so training volume could be compared. Subjects who

qualified for the study participated in a familiarization session in which the study was explained to the participants and informed consent was obtained. After the familiarization session, subjects were matched for bodyweight, years of training experience, and age and randomly assigned to one of three groups: 1.) KA at manufacturer’s ROCK inhibitor recommended doses (KA-L, 1.5 g/d for 28-days); 2.) KA at creatine equivalent loading (4 x 5 g/d for 7-days) and maintenance (5 g/d for 21-days) doses as CrM (KA-H); or, 3.) CrM at normal loading (4 x 5 g/d for 7-days) and maintenance doses (5 g/d for 21-days). Table 1 Overview of Study Design Familiarization and Entry Baseline

Day 0 Loading Phase Day 7 Maintenance Phase Day 28 Familiarization session 4-Day Diet History 4-Day Diet History 4-Day Diet History Informed Consent Form Muscle Biopsy Submit Training Log Submit Training RG7112 nmr Log Demographic Form Fasting Blood Sample Body Weight Muscle Biopsy Muscle Biopsy Health History Form Body Water (BIA) Fasting Blood Sample Fasting Blood Sample Exercise History Form DEXA Body Composition Body Weight Body Weight 4-day Dietary History 1 RM Leg Press Body Water (BIA) Body Water (BIA) General Exam to Vistusertib cost Determine Qualifications to Participate in Study 1 RM Bench

Press DEXA Body Composition DEXA Body Composition Height and Body Weight Wingate Anaerobic Capacity Test Wingate Anaerobic Capacity Test 1 RM Leg Press Practice Wingate Anaerobic Capacity Test Loading Phase of Supplementation Begins Low-Dose Maintenance Phase of Supplementation Begins 1 RM Bench Press Randomization into one of three groups (CrM, KA-L, KA-H) Maintain Training Log   Wingate Anaerobic Capacity Test Instructions for Supplementation       Participants Apparently healthy resistance-trained males with no self-reported recent history of creatine supplementation were recruited to participate in this study. Participants were not allowed to participate in this study if they had any metabolic disorder including known electrolyte abnormalities; heart Methane monooxygenase disease, arrhythmias, diabetes, thyroid disease, or hypogonadism; a history of hypertension, hepatorenal, musculoskeletal, autoimmune, or neurologic disease; if they were taking thyroid, anti-hyperlipidemic, hypoglycemic, anti-hypertensive, anti-inflammatory, or androgenic medications; or, if they had taken dietary supplements containing creatine within three months prior to the start of the study. Participants were recruited from the student population and from area fitness facilities. Participants completed demographic, health history and exercise history forms.

Finally, the second passivation layer on the top part of nanowire probe was etched selectively by blocking the rest of the probe, which was wrapped with polymethyl methacrylate. This anisotropic wet etching method makes the nanowire probe have a suitable structure for intracellular recording (shown in Figure 2d). The electrical properties of the nanowire XAV-939 chemical structure probes were characterized by measuring the cyclic voltammograms (CV) (Additional file 1: Kinase Inhibitor Library Figure S5 of supplementary data). CVs were measured with a Pt counter electrode and Ag/AgCl was used as a reference electrode. No decrease of current after a small peak

was observed in our nanoelectrode. Such a behavior is common in nanosize electrodes since analytes diffuse according to hemispherical diffusion in electrodes,

which leads to a higher mass transport per unit electrode surface. The sigmoidal voltammograms, which show limiting current, are characteristic of radial diffusion to cylindrical ultramicroelectrodes. Assuming that the electrode is a cylindrical Z-IETD-FMK purchase shape, the limiting plateau currents can be determined according to the following equation [35]. (1) Here, n is the number of electrons transferred during the electrochemical process, F is Faraday’s constant, D and C are the diffusion coefficient and concentration of the electroactive species respectively, l and r are the length and radii of nanoelectrode, respectively, and t is time scale of the

CV experiment, which is represented by RT/Fv. The experimental limiting current value at our nanoelectrode is 4.5 nA, which is similar to the theoretical limiting current value (4.21 nA/μm). The probing of neural activity was carried out using a rat clonal GH3 pituitary cell line, which has a spontaneous action potential that is known to be stimulated by a thyrotropin releasing hormone [36]. As such, it is ideal to test the feasibility of old the nanowire probe for measuring neural activity without external stimulation to induce an action potential. Figure 3a is an SEM image of the vertical nanowire probe device before the culturing of the GH3 cells. Culturing was carried out with GH3 cells 2 days after cell plating. The standard bath solution consisted of 140 mM NaCl, 5 mM KCl, 2 mM CaCl2, 1 mM MgCl2, 10 mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES), and 10 mM glucose was applied continuously into the culturing bath through a gravity-fed perfusion system during recording. Measurements were carried out at 25°C. Figure 3b is an SEM image of GH3 cultured in the same location as that shown in Figure 3a by seeding the cells of passage 10. The white circles in Figure 3b indicate the sites where the vertical nanowire probes are positioned. The image clearly shows that the nanowire probes are covered with GH3 cells. The individual probing electrode was connected to the input of a buffer.

5 mL microfuge tube, air dried briefly and suspended in 200 μL TE buffer resulting in a DNA concentration of approximately 1 μg/μL. Sequencing and annotation Random and φ52237-sequence guided φX216 genome fragment clones were constructed by GW-572016 price restriction digest of purified φX216 genomic DNA with EcoRI, EcoRI + HindIII or AgeI and ligation with EcoRI, EcoRI + HindIII or SmaI digested pUC19 DNA [25], respectively, followed by

transformation of E. coli DH5α or GBE180 [26] using standard transformation protocols [27] and recovery of white colonies on LB plates containing 100 μg/mL ampicillin and 50 μg/mL 5’-bromo-4-chloro-3-indolyl-β-D-galactopyranoside (X-gal). φ52237-sequence-guided PCR amplicons were designed to close gaps and confirm fragment clone borders. Sequencing was accomplished using M13F and M13R primers, as well as φ52237-sequence guided primer walking of fragment clones and PCR amplicons using an ABI 3130xL Genetic Analyzer (Applied Biosystems, Carlsbad, CA) at the Colorado AR-13324 State University Proteomics and Metabolomics Facility. φX216 Illumina sequencing libraries were prepared using the TruSeq DNA Sample Preparation Kit v2, (Illumina, San Diego, CA), following the manufacturer’s instructions. Phage DNA was fragmented to a range of 300–400 bp using a Covaris acoustic shearing device, (Covaris Inc., Woburn, MA) followed by 3′ adenylation and adapter ligation. Ligation eFT-508 chemical structure products were purified on an agarose gel and the DNA fragments

enriched via PCR. Fragmented Phage DNA was sequenced by high-throughput Adenylyl cyclase Illumina parallel sequencing using 100 bp mate-pair Illumina HiSeq 2000 reversible terminator chemistry. The library was run on 15% of a single lane. Reads were trimmed for quality and de novo short-read genome assembly was performed using the Velvet 1.1.05 sequence assembler algorithms with a hash length of

99 and a final graph with 3 nodes and n50 of 37412 nt [28]. Open reading frames were identified with GeneMark gene prediction software using a viral-optimized Heuristic approach [29]. Putative gene identification was conducted by sequence alignment with φ52237 (GenBank:DQ087285.2) [8] and individual open reading frames queried using the NCBI Basic Alignment Search Tool (BLAST). Genome annotation, mapping, sequence alignments, and comparative analyses were conducted using Gene Construction Kit v3.0 and Geneious Pro 5.4.6 bioinformatics software. The annotation map was created using Adobe Illustrator CS5. The final φX216 genome sequence has been deposited in GenBank under accession # JX681814. Acknowledgements Funding was provided by the Defense Threat Reduction Agency grant W81XWH-07-C0061. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Electronic supplementary material Additional file 1: φX216 host range, word document, Host range of φX216. Table of φX216 host range for 72 B. pseudomallei strains and other Burkholderia species.

J Exp Med 1988, 167:718–723.PubMedCrossRef 2. Morrison-Plummer J, Lazzell A, Baseman JB: Shared epitopes between Mycoplasma pneumoniae major adhesin protein P1 and a 140-kilodalton

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J 1996, 9:669–672.PubMedCrossRef 7. Ginestal RC, Plaza JF, Callejo JM, Rodríguez-Espinosa N, Fernández-Ruiz LC, Masjuán J: Bilateral optic neuritis and Guillain-Barré syndrome following an acute Mycoplasma pneumoniae infection. J Neurol 2004, 251:767–768.PubMedCrossRef 8. Stutman HR: Stevens-Johnson syndrome and Mycoplasma pneumoniae : evidence for cutaneous infection. J Pediatr 1987, 111:845–847.PubMedCrossRef 9. Yamane Y, Kawai C: A case of myocarditis caused by Mycoplasma pneumoniae . Jpn Circ J 1978, 42:1279–1287.PubMedCrossRef 10. Hakkarainen K, Turunen H, Miettinen A, Karppelin M, Kaitila K, Jansson E: Mycoplasmas and arthritis. Ann Rheum Dis 1992, 51:1170–1172.PubMedCentralPubMedCrossRef 11. Waites KB, Talkington DF: Mycoplasma pneumoniae and its role as a human pathogen. Clin Microbiol Rev 2004, 17:697–728.PubMedCentralPubMedCrossRef

12. Fernald GW, Collier AM, Clyde WA: Respiratory infections due to Mycoplasma pneumoniae in infants and children. Pediatrics 1975, 55:327–335.PubMed 13. Harrington CYTH4 LE, Hatton RD, Mangan PR, Turner H, Murphy TL, Murphy KM, Weaver CT: Interleukin 17-producing CD4+ effector T cells develop via a lineage distinct from the T helper type 1 and 2 lineages. Nat Immunol 2005,6(11):1123–1132.PubMedCrossRef 14. Park H, Li Z, Yang XO, Chang SH, Nurieva R, Wang YH, Wang Y, Hood L, Zhu Z, Tian Q, Dong C: A distinct lineage of CD4 T cells regulates tissue inflammation by producing interleukin 17. Nat Immunol 2005,6(11):1133–1141.PubMedCentralPubMedCrossRef 15. Nakae S, Nambu A, Sudo K, Iwakura Y: Suppression of immune induction of collagen-induced arthritis in IL-17-deficient mice. J Immunol 2003,171(11):6173–6177.PubMedCrossRef 16.

Microarray procedures Streptococcus mutans UA159 (NC004350) NimbleGen Genechip (4*72 K) whole-genome array DAPT cost was employed for transcriptional profiling in this study. The oligoarrays included 1949 S. mutans UA159 open reading frames with twelve 24-mer probe pairs (PM/MM) per gene, and each probe was replicated 3 times. The design also included random GC and other control probes. Array

images were scanned by Gene Pix® 4000B Microarray Scanner (Axon Instruments, Union City, CA, USA). Raw data were normalized using RMA algorithm by Roche NimbleScan software version 2.6. We used the average value of each replicate probe as the signal intensity for the corresponding gene, and all the values were log2 transformed for further analysis. The normalized data with PRIMA-1MET supplier annotation information was processed by combining several different R/Bioconductor packages. We conducted a non-parametric statistical method contained in the RanProd package to detect the EX 527 solubility dmso differentially expressed genes (DEG) [31]. With 100,000 permutation test, genes having a minimum 2-fold change with

the false discovery rate (FDR) smaller than 0.1 were considered as DEG, indicating a significant up- or down-regulation under hyperosmotic stress. For the pathway analysis, we firstly constructed the whole S. mutans UA159 pathway database based on the KEGG Pathway. Then gene set enrichment analysis (GSEA) was used to determine the pathways that changed significantly in response to hyperosmotic stress [32, 33]. The microarray results were further validated by quantitative RT-PCR for selected genes (see Additional file 3 for detailed primer sequences for qPCR). Quantitative RT-PCR assays were performed using a SYBR Green reverse transcription-PCR kit (TaKaRa, Dalian, China) according to the manufacturer’s instructions. Statistical analysis We used Student’s T-test to compare the non-treated control out groups with treatment groups. All statistical procedures

were conducted by R software [34]. Data were considered significantly different if the two-tailed P-value was < 0.05. Microarray data accession All the microarray raw data have been submitted to the NCBI Gene Expression Omnibus database under the accession number GSE47170 (http://​www.​ncbi.​nlm.​nih.​gov/​geo/​query/​acc.​cgi?​acc=​GSE47170). Acknowledgements This work was supported by National Natural Science Foundation of China (grant number: 81170959), Doctoral Fund of Ministry of Education of China (grant number: 20120181120002) and National Natural Science Foundation of China (grant number: 81200782). The authors would like to thank Arne Heydorn from Section of Molecular Microbiology, the Technical University of Denmark, for proving image-processing software COMSTAT. Electronic supplementary material Additional file 1: Heat map of different expressed genes of Streptococcus mutans UA159 in response to short-term hyperosmotic stress. Transcript enrichment is encoded in the heat map from low (blue) to high (red).