(A) CP-AP concentrations in serum specimens of healthy controls (

(A) CP-AP concentrations in serum specimens of healthy controls (HC), inflammatory controls (IC) and tumor patients (TP). In the box plot the central box represents the values from the lower to upper quartile (25 to 75 percentile). The middle line represents the median. The horizontal

line extends click here from the minimum to the maximum value. P-values of the Mann–Whitney test are indicated. (B) ROC-AUC calculation for separation of tumor patients (TP) from healthy controls (HC) (left graph), tumor patients (TP) from inflammatory controls (IC) (middle graph) and healthy controls from inflammatory controls (IC) (right graph). Discussion The dysregulation of protease activity plays an important role for the initiation and progression of malignant disease [1, 4]. Tumor-associated proteases like matrix metalloproteases, cathepsins, kallikrein related peptidases and members of the plasminogen activator system are Everolimus concentration secreted into the bloodstream and might be candidates for functional protease profiling (for review see [20]). Specifically, the tumor-associated protease cancer procoagulant is secreted from numerous malignancies including colorectal cancer into the bloodstream [21]. Under in vivo conditions this can cause paraneoplastic

coagulopathy throughout cleavage and activation of the coagulation factor X heavy chain (P00742) [22]. The reporter peptide CP-RP comprises the cleavage site WKPYDAAD that is part of the coagulation factor X and is preferably cleaved in serum specimens of tumor patients [8]. Adding reporter peptides to selleck chemicals serum specimens enables the monitoring of tumor-related proteolytic activity for diagnostic use [7–9, 23, 24]. Furthermore, reporter peptide spiking offers major advantages over native MS-based peptide profiling concerning the standardization of preanalytical variabilities [6, 11]. The main focus of our present work was to optimize functional protease profiling with respect to simplified sample preparation and increased inter-day reproducibility to make it amenable as a laboratory assay for routine diagnostic use. Recently, a sample

clean-up with trichloroacetic acid (TCA) has been described that showed a sufficient recovery for peptides with a molecular weight of less than 3000 Da [25]. Furthermore, diglyceride the LC-MS technique is the method of choice for the reproducible quantification of small molecules like peptides in clinical specimens [26], and accordingly this technology was selected for assay development. Even at low CP-AP concentrations of 0.4 μmol/L the extracted ion chromatogram of CP-AP with m/z 515.795 shows only one single peak (see Figure 1) and this excellent signal to noise ratio makes quantitative LC/MS analyses amenable [27, 28]. Recently, criticism has been raised against functional protease profiling and it has been suggested to characterize the proteolytic activity in more detail [29].

The pellet obtained was suspended in Buffer A plus 0 5% Triton X-

The pellet obtained was suspended in Buffer A plus 0.5% Triton X-100 (Buffer B) at room temperature. After 1 h, the suspension was ultracentrifuged (161,000 × g, 1 h), and the supernatant obtained was stored at 4°C. The cell-free extract solubilized

(about 120 mg) was applied to a column of TALON metal affinity resin (TaKaRa Bio, Inc. (Shiga, Japan); 10 × 15 cm). The column was equilibrated with Buffer B at a flow rate of 0.5 ml/min, and washed successively with Buffer B (90 ml), Buffer B plus 10 mM Imidazole (16 ml), Buffer B plus 20 mM Imidazole (16 ml), and Buffer B BIBW2992 price plus 50 m M Imidazole (4 ml). The adsorbed protein was eluted with Buffer B plus 250 mM imidazole (20 ml). The elution was collected with a Bio-collector (ATTO, Tokyo. Japan, 2 ml/tube), and the protein concentration MLN2238 order was measured with a RC DC Protein assay kit (Bio-Rad Laboratories, Inc., Hercules, CA, USA). The fractions containing the D-lactate dehydrogenase were dialyzed against two 1-l portions of Buffer A for 4 and 12 h, and stored at 4°C. Comparative transcriptome analysis using DNA microarrays Generation of C. glutamicum whole-genome DNA microarrays, total RNA preparation, synthesis of fluorescently labelled cDNA, microarray hybridization, washing, and statistical data analysis were performed as described previously [35–38]. Genes exhibiting mRNA levels that were significantly changed (P ≤ 0.05 in Student’s t test) by at least a factor of 2.0 were determined

in three DNA microarray experiments performed with RNA isolated from three independent cultures. The processed and normalized data have been deposited in the NCBI’s Gene Expression Omnibus and are accessible under the accession number Bcr-Abl inhibitor GSE25704. Results Cg1027 encodes D-lactate dehydrogenase The C. glutamicum ATCC 13032 gene cg1027 was annotated to code for D-lactate dehydrogenase [39] as the deduced protein shows similarities to FAD/FMN-containing dehydrogenases encoded by the cluster of orthologous genes COG0277. The deduced

protein contains the Selleck AZD1390 conserved domain PRK11183, and the domain (aa 279-570) was similar to membrane-binding D-lactate dehydrogenases belonging to the protein family pfam09330. In order to determine whether the gene product of cg1027 is indeed active as D-lactate dehydrogenase, the gene was cloned into pET14b, and the hexahistidine-tagged protein was purified from E. coli BL21 (DE3) harboring pET14b-dld. Quinone-dependent D-lactate dehydrogenase activity was detected by using 2,6-dichloroindophenol as an electron acceptor. The optimum assay conditions were observed in a 100 mM potassium phosphate buffer at a pH of 7.0 and a temperature of 45°C. Subsequently, Dld activity was assayed at 30°C, the optimal temperature for growth of C. glutamicum. The enzyme showed Michaelis-Menten kinetics with D-lactate as the substrate and it was determined that 0.61 mM of D-lactate resulted in half maximal enzyme activity. The observed V max was 73.5 μmol mg-1.


This BB-94 cost approach of growth curve synchronization has several advantages over sampling a system at different times. Firstly, the endpoint measurements can all be performed at the same time, thereby decreasing experimental variability. Secondly, efficiency will be improved compared to processing multiple samples at different times. Thirdly, no invasive sampling is necessary and the method requires no constant vigilance or presence. Finally, as we discuss throughout the paper, it allows measuring the division rate of cells

directly from optical density with very high precision. We exemplify the growth curve synchronization method by analyzing rhamnolipid secretion by the bacterium Pseudomonas aeruginosa. P. aeruginosa is an opportunistic human pathogen found in long-term, often terminal, infections in cystic fibrosis patients and various nosocomial infections occurring in immunocompromized JQEZ5 price patients [2–9]. Rhamnolipids are among the predominant virulence factors of P. aeruginosa [9, 10]. These glycolipid surfactants are involved in the formation and maintenance of biofilms, cytolysis of polymorphonuclear Tozasertib molecular weight leukocytes (PMNs) and swarming motility ([8, 11]; reviewed in [12]). Their synthesis is regulated by quorum sensing, a mechanism for cell density-dependent

gene regulation. As such, rhamnolipid secretion in P. aeruginosa is a valuable model system to investigate how pathogenic bacteria coordinate population-wide traits at the molecular level [13]. The rhamnolipid quorum-sensing regulation consists of at least two hierarchical systems governed by two different autoinducers [14–23].

These two systems, called rhl and las, share a common motif. An autoinducer synthase (RhlI and LasI) synthesizes Florfenicol the autoinducer (N-butyryl-L-homoserine lactone or C4-HSL and N-(3-oxododecanoyl)-L-homoserine lactone or 3O-C12-HSL), which binds to its cognate transcription factor (RhlR and LasR) that, in turn, up-regulates the autoinducer synthase in a positive feedback. LasR controls expression of RhlR, and thereby the las system is hierarchically above rhl. The rhl system induces expression of rhlAB, resulting in rhamnolipid production [24]. In spite of this knowledge, the rhamnolipid system has puzzled microbiologists because it does not behave like the paradigm of quorum sensing [13, 25, 26]. In either rhlI – or lasI – bacteria, adding autoinducers to the growth media does not induce rhamnolipid secretion from the outset of the culture, indicating there is at least one other factor regulating rhlAB expression [13]. Here we illustrate our growth curve synchronization method by integrating high-resolution spectrophotometric measurements of cell density and gene expression with endpoint rhamnolipid quantification to produce multi-measurement time series of the latter.

Activated CheY (CheY-P) interacts directly with the motor of the

Activated CheY (CheY-P) interacts directly with the motor of the flagella to control swimming direction. The dephosphorylation of CheY-P occurs spontaneously, only in enterobacteria this reaction is accelerated by the phosphatase CheZ. For adaptation, CheB and its antagonist CheR remove or add methyl groups to the receptors, buy Salubrinal respectively. In R. centenaria, the two central components of the chemotactic signal transduction cascade, namely CheA and CheY, are present as the fusion protein Rc-CheAY located in the first chemotactic operon [17], a situation that is also observed in Selleckchem Combretastatin A4 Helicobacter [18]. Whereas the role

of the CheY-domain of the CheAY protein in H. pylori seems to be a phosphate sink, in R. centenaria, the function of Rc-CheAY remains still unclear. While Che proteins are generally involved in chemotactic responses, they were also shown to affect the phototactic response in R. centenaria as demonstrated by the analysis of many che mutants [19]. In the last decade, bacterial photoreactive proteins like phytochromes, previously thought to be a unique feature in plants, have been identified as photoactive yellow proteins (Pyp) and have now been extensively studied in a variety of eubacterial species (for review see [20, 21]). For R. centenaria, a Pyp-like protein, Ppr, was described in 1999 by Bauer and colleagues

[22]. The large fusion protein Ppr consists of three learn more functional domains, an N-terminal Pyp domain with the cinnamic acid chromophore, the central phytochrome-like

bilin attachment domain Bbd and the C-terminal histidine kinase domain Pph which autophosphorylates Resminostat an essential histidine residue [22]. Although some Pyp proteins have been crystallized and biophysically characterized in great detail (reviewed by [21]), no distinct physiological role could be attested to these unique proteins. A Ppr-deletion mutant lacking amino acid residues 114-750 did not show any alterations in phototactic behaviour, instead exhibited a strongly deregulated expression of the chalcone synthase gene suggesting a regulatory function in the polyketide synthesis [22]. Although there is no obvious direct involvement of Ppr in the phototactic or scotophobic reaction, an interaction with the chemotactic signal transduction components is plausible to regulate general phosphorylation levels or transduce phosphoryl groups to a yet unknown light-dependent signal transducing protein. We therefore analysed whether the Ppr protein and in particular its phosphorylating kinase domain Pph interacts with the Rc-Che proteins. Results The chemotactic response of E. coli is inhibited by the expression of Ppr The chemotactic network in E. coli is very sensitive to alterations in the expression level and stoichiometry of the chemotactic proteins Ec-CheW [23, 24] and Ec-CheA [25] as well as the MCP receptors [26, 27].

Clin Endocrinol (Oxf) 2010, in press 20 Delarue J, Matzinger O,

Clin Endocrinol (Oxf) 2010, in press. 20. Delarue J, Matzinger O, Binnert C, Schneiter P, Chiolero R, Tappy L: Fish oil prevents the adrenal activation elicited by mental stress in healthy men. Diabetes Metab 2003, 29:289–295.CrossRefPubMed 21. Couet C, Delarue P, Autoine JM, Lamisse F: Effect of dietary fish oil on body mass and basal fat oxidation in healthy adults. Int J Obes 1997, 21:637–643.CrossRef 22. Hill AM, Buckley JD, Murphy KJ, Howe PR: Combining fish-oil supplements with regular aerobic exercise improves body composition and cardiovascular disease risk factors. Am J Clin Nutr 2007, 85:1267–1274.PubMed

23. Thorsdottir I, Tomasson H, Gunnarsdottir I, Gisladottir E, Kiely M, Parra MD, Bandarra NM, Schaafsma G, Martinez JA: Randomized trial of weight-loss-diets for young adults varying in fish and fish oil content. Int J Obes (Lond) 2007, 31:1560–1566.CrossRef 24. Dempster P, selleck chemical SYN-117 Aitkens S: A new air displacement method for the determination of human body composition. Med Sci Sports Exerc 1995, 27:1692–1697.PubMed 25. Siri

WE: Body composition from fluid spaces and density: analysis of methods. In Techniques for measuring body composition. Edited by: Brozek J, Henschel A. Washington, DC: National Academeny of Sciences, National Research Council; 1961:223–244. 26. Zuntz H: Pflugers Arch Physiol. 1901, 83:557.CrossRef 27. Hellhammer DH, Wust S, Kudielka BM: Salivary cortisol as a biomarker in stress research. Psychoneuroendocrinology Succinyl-CoA 2009, 34:163–171.CrossRefPubMed 28. Gallagher D, Belmonte D, Deurenberg P, Wang Z, Krasnow N, Pi-Sunyer FX, Heymsfield SB: Organ-tissue mass measurement allows modeling of REE and metabolically active tissue mass. Am J Physiol 1998, 275:E249–258.PubMed

29. Illner K, Brinkmann G, Heller M, Bosy-Westphal A, Muller MJ: Metabolically active components of fat free mass and resting energy expenditure in nonobese adults. Am J Physiol Endocrinol Metab 2000, 278:E308–315.PubMed 30. Rodriguez G, Moreno LA, Sarria A, Pineda I, Fleta J, Perez-Gonzalez JM, Bueno M: Determinants of resting energy expenditure in obese and non-obese children and adolescents. J Physiol Biochem 2002, 58:9–15.CrossRefPubMed 31. Bosy-Westphal A, Eichhorn C, Kutzner D, Illner K, Heller M, Muller MJ: The age-related decline in resting energy expenditure in humans is due to the loss of fat-free mass and to alterations in its metabolically active components. J Nutr 2003, 133:2356–2362.PubMed 32. Byrne HK, Wilmore JH: The effects of a ATM Kinase Inhibitor 20-week exercise training program on resting metabolic rate in previously sedentary, moderately obese women. Int J Sport Nutr Exerc Metab 2001, 11:15–31.PubMed 33. Horner NK, Lampe JW, Patterson RE, Neuhouser ML, Beresford SA, Prentice RL: Indirect calorimetry protocol development for measuring resting metabolic rate as a component of total energy expenditure in free-living postmenopausal women. J Nutr 2001, 131:2215–2218.PubMed 34.

Here, we demonstrate our extended effort to extensively study the

Here, we demonstrate our extended effort to extensively study the structural properties and, in particular, the photocatalytic application of these hybrid nanocatalysts. Methods A modified microwave method was used to synthesise the TiO2/MWCNTs hybrid nanocatalysts. Initially, a 3.5-cm hole was drilled through the top of a household microwave oven. A reflux condenser was subsequently installed in the microwave oven to enable continuous synthesis at ambient pressures. Since the microwave has a wavelength of 12 cm, there will be no escaped radiation through the hole. As additional protection purpose, the microwave

was operated inside a fume hood. Commercial MWCNTs (Cheap Tubes Inc., Brattleboro, VT, USA) with an outer buy Torin 2 diameter of 10 to 30 nm, an inner diameter of 5 nm, a surface area of 110 m2/g and lengths up to 50 μm were used in this work. Due to electrostatic interactions and van der Waals forces between the individual nanotubes, the MWCNTs exhibit a strong tendency to agglomerate. This agglomeration

leads to poor solubility of the MWCNTs in most aqueous and organic solvents. Thus, to achieve a stable aqueous suspension of MWCNTs, functionalisation processes are necessary due to the presence of a large ISRIB in vivo amount of functional groups on the nanotubes’ surface. The presence of these functional groups on the MWCNTs’ surface imparts negative charges and thus generates repulsion forces, which inhibit agglomeration. These negative charges can also function as anchor sites and thereby enable the in situ attachment of synthesised nanoparticles onto the MWCNTs’ surface. For this purpose, the MWCNTs were first functionalised by being

sonicated for 3 h in a 65% TPX-0005 solution of concentrated HNO3. The suspended MWCNTs were then placed in the modified microwave oven (Sharp model R-369 T) and irradiated for 20 min at a power of 550 W. Afterwards, the product was rinsed with deionised water six times and then completely dried at 80°C. old The MWCNTs were denoted as functionalised MWCNTs (f-MWCNTs) after this process. The surface areas of the f-MWCNTs dramatically increased to 357.6 m2/g after the functionalisation process. Greater MWCNT surface area recorded after functionalisation has been associated with the increase of functional groups on the nanotube surface [39]. Preparation of TiO2/MWCNTs nanocatalysts involved the dispersion of f-MWCNTs in ethanol (pH = 2) and sonicated for 1 h. Then, approximately 561 μL of titanium isopropoxide (TTIP) was added dropwise to the suspension over a period of 20 min under vigorous stirring. Notably, under acidic conditions, the TiO2 surface contains positive charges due to the presence of ≡Ti-OH2 + groups [40], which enhance the adhesion characteristics on the MWCNTs’ surface. The amount of TTIP precursor represented a TiO2/f-MWCNT weight ratio of 50%.

g open water, estuary, sediments), and may lead to the local eme

g. open water, estuary, sediments), and may lead to the local emergence of better adapted types [51, 52]. For

example STs that were frequently identified within our study were either present in the North Sea or the Baltic Sea but not in both. Thus the natural subdivision of North Sea and Baltic Sea seems to represent different habitats to which different strains may be better adapted to. Possibly the differences of ST-distribution in Sri Lankan and Ecuadorian prawn farms could be based on differing structures within shrimp farms, e.g. approx. 50% of the purchased post larvae in Sri Lankan shrimp ponds were obtained from only four vendors (one vendor supplies 24.1% of ponds), whereas in Ecuador all farms we included PLX-4720 chemical structure in our study purchased their post larvae from individual vendors ([51], unpublished data). In single cases we were able to trace individual STs along the food chain: from seafood producing areas like Sri Lanka and Ecuador up to the retail level in Germany. Additional analysis of the genetic diversity on smaller geographical scales (e.g. on a single farm, in a distinct bight) may help to understand

if the singletons STs (or pSTs) represent locally and environmentally adapted types with a clonal structure. On the other hand low scale strain communities could also be diverse due to the introduction of new strains or genetic exchange within present types and mutational events. Clusters of STs were identified by UPGMA that were dependent on the geographic origin and represented the local distribution of STs. Similarly, González-Escalona et al. RAD001 ic50 observed a distinct cluster of strains isolated from patients after the consumption of raw oysters from the U.S. Pacific coast [13]. But in our data, multiple clusters per continent were identified and the distribution of STs was independent of the geographic origin (e.g. STs of all continents are scattered over the whole UPGMA tree). On peptide level the loss

of geographical clusters of pSTs in Histidine ammonia-lyase the corresponding UPGMA tree was due to the global dissemination of pSTs. Like Osorio et al. showed, on peptide level nearly all pSTs were grouped in one cluster [28]. By comparing the results obtained by UPGMA analysis of MLST and AA-MLST data, clusters on nucleotide level were not always found on peptide level (Figures 3A and B). But all STs that form a CC or doublet were characterized by the same pST (CC410 and doublet ST246-ST56 were pST1; doublet ST760-ST412 was pST6). This showed that both typing https://www.selleckchem.com/products/RO4929097.html schemes provided different clustering results due to the decreased resolution of the AA-MLST approach, but with concordance in grouping CCs and doublets emphasizing the high degree of genetic similarity found within these groups. In the case of using a sequence based UPGMA tree no additional information was gained by application of AA-MLST analysis. Population structure of V.

This is necessary

because the amount of oleic acid affect

This is necessary

because the amount of oleic acid affects MNC formation. Steric repulsion among the hydrocarbon tails of oleic acid on individual MNPs impacts assembly capability of individual MNPs. To modify the amount of oleic acid on the MNPs, the MNPs were dissolved in n-hexane Endocrinology inhibitor and ethanol was added to the solution to remove part of the oleic acid coating. Finally, three samples of PMNPs were successfully obtained from the precipitates [25, 26], each coated with different oleic acid amounts: 19 (low PMNPs, LMNPs), 33 (medium PMNPs, MMNPs), and 46 (high PMNPs, HMNPs) wt.% (Figure 2b). To investigate the effect of primary ligand on MNCs, the interactions of oleic acid molecules on the surface of MNPs were analyzed through derivative weight curves of the three samples of PMNPs (Figure 2c). These PMNPs showed three derivative peaks positioned between 25°C and 550°C [28–30]. The first peak positioned at approximately 250°C (Figure 2c, i) was due to the removal of free oleic acid molecules surrounding the MNPs (Figure 2d,

i), consistent with the derivative peak of pure oleic acid (Additional file 1: GSK2118436 in vitro Figure S2). The second peak positioned at approximately 350°C (Figure 2c, ii), which was close to the boiling temperature of oleic acid, indicated bilayered oleic acid molecules with hydrophobic interactions between hydrocarbon tails (Figure 2d, ii). The third peak at approximately 450°C (Figure 2c, iii) corresponded to oleic acid molecules covalently bound to MNPs (Figure 2d, iii). The characteristic peaks of the oleic acid-MNP conjugates from asymmetric and symmetric COO− stretches of oleic

acid (1,630 Bucladesine in vitro and 1,532 cm−1) were confirmed by FT-IR spectroscopy (Additional file 1: Figure S3 and Table S1) and were categorized as a chelating bidentate complex: peak separation as Evodiamine 98 cm−1 = 1,630 to 1,532 cm−1 (Additional file 1: Table S2) [30, 31]. The derivative weight curve of an iron-oleate precursor used for MNP synthesis also agreed with the derivative peaks of PMNPs (Additional file 1: Figure S4). From these results, it was determined that LMNPs contained mostly surface-bound oleic acid molecules showing a sharp peak approximately 450°C (Figure 2c, red line). Increased oleic acid in MMNPs formed a surface bilayer, which showed as an additional derivative peak at approximately 350°C (Figure 2c, blue line). The appearance of a sharp peak at approximately 250°C in HMNPs represented excess free oleic acid molecules (Figure 2c, black line). Therefore, we expected that (1) LMNPs were more likely to agglomerate and form large dense MNCs, (2) MMNPs would undergo less self-assembly and form smaller MNCs compared with LMNPs, and (3) excess free oleic acid in HMNPs would disrupt the assembly of individual MNPs to form MNCs. Following primary-ligand modulation, PMNPs were then emulsified with the nanoemulsion method, using polysorbate 80 as a secondary ligand to fabricate MNCs.

1% sodium azide and 0 05 mM EDTA and resuspended in the same buff

1% sodium azide and 0.05 mM EDTA and resuspended in the same buffer to a density of 5 × 106 cells/ml. The following anti-mouse monoclonal antibodies directed against surface antigens were used: TcR1-FITC (clone GL3) from AbD Serotec and CD19-PE-Cy5.5 (clone 6D5), CD3-APC (clone 145-2C11),

CD45-FITC (clone 30-F11), CD16/32-PE (clone 93) and CD14-FITC (clone Sa2-8) from eBioscience. Before the flow cytometry, the isolated lymphocytes were incubated with the appropriate antibodies for 30 min, washed twice in PBS and analyzed by FACSCalibur™ (BD Biosciences) equipped with a 488 nm argon-ion laser and a 633 nm diode laser. At least 105 cells were analyzed and data analyses of gated lymphocytes positive for CD45 were performed using CELLQuest™ Pro software (BD Biosciences). γδ T-lymphocytes

were identified in a single TcR-specific staining. CD19-positive B-lymphocytes and CD3-positive T-lymphocytes, and CD4 and CD8 Th- and Tc-lymphocytes, were each characterized selleck products by separate two-colour analysis. Finally, the CD14 and CD16 positive cells out of CD3 and CD19 double negative were quantified using a four-colour analysis. Real time PCR Total RNA was extracted from caecal wall samples using the RNeasy Lipid Tissue Kit (Qiagen). Resulting RNA was eluted with 50 μl RNase-free water and used immediately in reverse transcription using M-MLV reverse transcriptase (Invitrogen) and oligo-T primers. The resulting cDNA was purified by the QiaPrep PCR Purification kit (Qiagen) and used as a template for quantitative PCR. mRNA expression rates of TNFα, CP673451 solubility dmso IL-12p40, IL-18, IFNγ and iNOS were determined using the QuantiTect™ SYBR® Green RT-PCR Kit (Qiagen) with β-actin mRNA as a reference. Primers used for the RT-PCR are listed in Table 4. The threshold cycle values (Ct) of gene of interest were first Selleck OICR-9429 normalised to the Ct value of actin

reference mRNA (ΔCt) and the normalised mRNA levels were calculated as 2(-ΔCt). The normalised mRNA levels of a particular cytokine were then used for t-test comparisons between the infected and non-infected animals and are also given in figures as “”actin”" units. Table 4 List of primers used for the quantification of gene expression by real time RT PCR. primer sequence 5′-3′ length (bp) Reference TNFαFor CATCTTCTCAAAATTCGAGTGACAA 175 [34] TNFαRev TGGGAGTAGACAAGGTACAACCC     IL-12p40For GGAAGCACGGCAGCAGAATA 180 [34] IL-12p40Rev Atezolizumab AACTTGAGGGAGAAGTAGGAATGG     IL-18For CAGGCCTGACATCTTCTGCAA 105 [34] IL-18Rev TCTGACATGGCAGCCATTGT     IFNγFor AACAGCAAGGCGAAAAAGGA 92 this study IFNγRev GTGGACCACTCGGATGAGC     iNOSFor CAGCTGGGCTGTACAAACCTT 95 [34] iNOSRev CATTGGAAGTGAAGCGTTTCG     β-actinFor CTTTGCAGCTCCTTCGTTG 150 this study β-actinRev ACGATGGAGGGGAATACAGC     Statistical analysis Data were evaluated by parametric two-sample, equal variance, t-test and non-parametric Mann-Whitney test comparing the experimental groups either to the non-infected control mice or to the mice infected with the wild type S. Enteritidis.

Mol Microbiol 2012, 83:759–774 PubMedCentralPubMedCrossRef

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