014 mg/kg 1,34 parathyroid hormone, and the estrogen (E) group receiving 15 g/day food, so the average E intake was 0.5 mg E/day corresponding to 0.325 mg free 17-β-estradiol, and an untreated non-OVX group was added as sham-operated group. The experimental procedures were approved by the local ethics commission under German animal Gefitinib protection law (permission from 11.03.1998, AZ: 509.42502/01-02.98 Bezirkregierung Braunschweig). Eight weeks before starting the drug treatments, bilateral ovariectomy was performed. After 5 weeks of drug treatments,

the rats were euthanized, and bilateral femurs were dissected free of soft tissue and then submitted to biomechanical and histomorphometric tests. Intravital fluorochrome labeling During the 35 days of drug treatment, animals were subcutaneously injected with four fluorescent agents (Merck, Darmstadt, Germany) to label the process of bone formation and restoration. The following fluorochromes were used: xylenol orange (90 mg/kg) on day 13, calcein green (10 mg/kg) on day 18, alizarin red (30 mg/kg) on day 24, and tetracycline (25 mg/kg) on day 35. The results of the fluorochrome labeling were analyzed quantitatively in the cross sections of femurs 11 mm distal from femoral head in the subtrochanteric region. Evaluation of

the changes and the localization of bone formation in the cortical surface was the aim of fluorochrome YAP-TEAD Inhibitor 1 clinical trial analysis. Biomechanical test During the breaking test, the actual strength was recorded every 0.1 mm during next the lowering of the stamp. The testXpert software

continuously recorded the force applied until total failure of the bone occurred. After the failure, the software program indicated the maximum load (F max) and the breaking strength. The breaking strength is the last measured point of the running graph and has no explanatory power. In the right–left comparison and the comparative bioassay, F max is the highest force that the femur can withstand. According to the method described in Stuermer et al. (2006), increases in elastic deformation (stiffness = elasticity) were calculated, and the transition point of elastic to plastic deformation was determined from the digital data [15]. This point represents the yield load of the bone. To determine this point, we calculated a regression line and the standard deviation (SD) with the individual data of the linear part of the graph. We defined the transition point of elastic to plastic deformation as a decrease of stiffness of more than twice the SD. X-ray examination of fracture mode Radiographs in the anterior–posterior and lateral view of all femurs tested in the comparative bioassay were taken. A special film (Kodak SR type 45) and a Faxitron fine-focus cabinet X-ray system (model 43855A; Faxitron X-ray System) with 40 kV were used.

As indicated in Figure 3a, the methyl group of vanillate cleaved by O-demethylase enters the methyl branch to form CO2 while generating reducing power that could be used to convert CO2 to CO. Twenty homologs were identified in the DCB-2 genome for the gene encoding a vanillate-specific O-demethylase corrinoid protein (odmA) while 15 were found

in Y51 [9, 19]. Figure 3 The Wood-Ljungdahl pathway and CO 2 fixation in D. hafniense DCB-2. (a) Key enzymes involved in the Wood-Ljungdahl Fulvestrant chemical structure pathway and the corresponding gene homologs are indicated. The pathway depicts the methyl branch (left) and the carbonyl branch (right) prior to forming acetyl-CoA. Reactions for the methyl group that is derived from vanillate demethylation are indicated with red arrows; DHB, 3,4-dihydroxybenzoate. FK506 molecular weight Homolog searches were performed by BLASTP with cutoff values of 1e-2 (E-value) and 30% identity in amino acid sequence. (b) Autotrophic cell growth of D. hafniense DCB-2 as measured by total number of the cell per ml culture. M. thermoacetica grows autotrophically on CO2 and H2 using the Wood-Ljungdahl pathway, but since no ATP is gained from substrate-level phosphorylation by this pathway, anaerobic respiration

is implicated [16]. Establishment of a proton gradient through formate hydrogenlyase activity was postulated as one of potential mechanisms for energy generation [16]. Since DCB-2 has genes for the same pathway for CO2 fixation and for formate hydrogenlyase (Dhaf_4269-4271), we tested its ability to grow solely on CO2 Methamphetamine and H2. While DCB-2 grew under this condition compared to a no-H2 control (Figure 3b), the growth was not as robust as M. thermoacetica run in parallel. In addition, the growth results also indicate that CO was metabolized, presumably oxidized to form H+ and CO2 by CO dehydrogenase encoded by four gene copies (Figure 3a). The CO2 would then enter the methyl branch of the Wood-Ljungdahl pathway to produce a methyl group. In the photosynthetic

bacterium Rhodospirillum rubrum, CO induces CO dehydrogenase (CooS) and CO-tolerant hydrogenase (CooF), which allows cell growth in a CO-dependent manner in the dark [20]. By BLAST search we identified a gene similar to cooF (E value of 2e-49) located within a twelve-gene operon (Dhaf-4277-4288). The operon also encodes gene homologs for E. coli hydrogenases 3 and 4, both of which are part of formate hydrogenlyase complexes [21]. Similar to NADH dehydrogenase and to the CooF of R. rubrum, E. coli hydrogenase 4 has been implicated in proton translocation [21]. Other genes in the operon include two sporulation-related genes, ygfCD, and genes for phosphate starvation-inducible protein PhoH, a phosphohydrolase, and a diacylglycerol kinase. Energy metabolism Electron transport chain Ubiquinone and menaquinone in bacteria are lipid-soluble molecules that shuttle electrons between the membrane proteins in the electron-transport chain.

Typical fatigue behavior was seen in SHAM-ovariectomized as well as in ZOL-treated, ovariectomized rats. Fatigue properties, trabecular microarchitecture, and cortical thickness were similar in both groups. Previously, we showed that static compressive behavior was also similar in L3 vertebrae

of the same groups of rats [13]. Altogether, this suggests that ZOL treatment of ovariectomized rats results in the same vertebral bone mass and structure as SHAM, ovariectomized rats, as well as the same vertebral static and fatigue properties. For all vertebrae, force–displacement curves displayed typical fatigue behavior characterized by decreasing secant stiffness, increasing hysteresis, selleck chemical and increasing nonlinearity. This agrees with compressive, fatigue behavior previously reported for cortical and trabecular bone specimens [27, 31–33]. Also, the strong linear correlation between the log steady-state Cilomilast manufacturer creep rate and the log time to failure agrees with the literature [32, 33], which indicates the validity of the test. This also indicates that the integral fatigue behavior of cortical and trabecular bone in rats is similar to the two bone compartments assessed separately. We found an average apparent strain at failure of about 4% for both groups,

which is just slightly higher than the 3.4% and 2.8% reported for, respectively, human and bovine trabecular bone [31, 33]. Samples that did not fail during the test were removed from further analysis and showed a decreasing rather than an increasing Buspirone HCl apparent strain range per cycle during the test accompanied by an increasing secant stiffness. This behavior suggests that artifacts were present in these tests [41, 42], possibly due to vertebral ends that were not perfectly parallel. In this case, when the force range, leading to 0.75% apparent strain, was determined at the start of the test, the actual

area bearing the load would be smaller than the total bone area. During the test, the area bearing the load would then be compressed, resulting in the same load being born by the area of the whole vertebra and thus in lower strains. Improving the sawing procedure and specimen fixation in the loading device could possibly reduce the rate of exclusion of samples. The fatigue behavior in these whole vertebrae was comparable to the fatigue behavior found in studies on cortical and trabecular bone, though no fatigue data on rat bone are available. Although not determined in our study, it would be interesting to study whether failure starts in the cortical or trabecular bone. Most of the fatigue properties were unrelated to cortical or trabecular bone morphology, with the exception of weak relationships between trabecular bone morphology and apparent strain at failure.

With the advancement of DNA-based biosensors and automation for bacterial detection, enrichment broths could be screened for the presence of Campylobacter spp. in a shorter time, with greater sensitivity and without the generation of any microaerobic condition. In addition, food microbiology laboratories interested in establishing techniques

for the isolation of Campylobacter from retail meat will have access to a cost-effective enrichment procedure without the need to invest in systems to generate microaerobiosis. Reference documents from the FDA and FSIS USDA should eventually be updated to provide for an alternative, simplified protocol that yields similar number of find more Campylobacter positive samples as the current

reference protocols. Methods Sample preparation, incubation and Campylobacter isolation Retail broiler meat samples (total = 108 samples; 49 breasts and 59 thighs) were purchased from local stores (Auburn, AL) from April 2009 to October 2010. Samples were tested in batches of three to five samples per week. Each meat package was considered one sample, and from each package ~1-inch pieces were cut aseptically and mixed thoroughly. For all samples, 25 g of meat was weighed two times (two subsamples) in individual, sterile Whirl-Pak® (Nasco, Fort Atkinson, WI). Each subsample was enriched in 100 ml of Bolton’s broth (with antimicrobial supplements) and 5% (v/v) of lysed horse blood [17]. The control subsamples (microaerobic

Selleckchem Midostaurin subsamples) were incubated in anaerobic jars gassed with a microaerobic gas mix (85% N2, 10% CO2, 5% O2; Airgas, Radnor, PA) using the evacuation-replacement system MACSmics Jar Gassing System (Microbiology International, Frederick, MD). The other subsamples (aerobic subsamples) were incubated without the addition of microaerobic gas mix, by closing the bags after removing the remaining air manually. All subsamples were incubated at 42°C for 48 h. After incubation and for all subsamples, 0.1 ml of the enriched broth was transferred many to modified charcoal cefoperazone deoxycholate agar [10] through a 0.65 μm membrane filter as described elsewhere [33]. All agar plates were incubated under microaerobic conditions at 42°C for 48 h. Presumptive Campylobacter colonies were observed under phase contrast microscopy (Olympus BX51, Olympus America Inc., Center Valley, P) for spiral morphology and darting motility. Presumptive isolates were stored at -80°C in tryptic soy broth (Difco, Detroit, MI) supplemented with 20% glycerol (v/v) and 5% (v/v) lysed horse blood for further analysis. Identification of presumptive Campylobacter isolates by mPCR assays Campylobacter isolates were recovered from frozen stocks by transferring to Brucella agar plates supplemented with 5% horse blood and through 0.6 μm membrane filters as described above. Plates were incubated at 42°C under microaerobic conditions for 24 h.

parahaemolyticus and the addition of MAPK inhibitors, SB203580 (5 μM), SP600125 (15 μM) or PD98059 (40 μM), as indicated. Results indicate mean ± SEM of three independent experiments.

*P < 0.05 vs cells co-incubated with bacteria in absence of inhibitor. Discussion The results of this study demonstrate that V. parahaemolyticus causes activation of MAPK in human intestinal epithelial cells and that this activation is linked to the cellular responses elicited by this bacterium. V. parahaemolyticus induced activation of each of the MAPK - find more JNK, p38 and ERK – in Caco-2 and HeLa cells (Figure 1 and 2). A mutant strain with a non-functional TTSS1 (ΔvscN1) did not cause MAPK activation, providing

the first evidence that TTSS1 is responsible for the activation of MAPK in epithelial cells in response to infection with V. parahaemolyticus (Figure 2). While the role of TTSS1 in ERK activation was difficult to observe in Caco-2 cells, differences in the activation of ERK in HeLa cells co-incubated with WT compared to ΔvscN1 bacteria were clearly high throughput screening evident. V. parahaemolyticus therefore now joins a select group of gram-negative pathogens that use TTSS effectors to activate MAPK signalling to promote pathogen infection. Given the important role MAPK play in controlling host innate immune responses and cell growth, differentiation and death, they are commendable targets for pathogenic effectors. While several pathogens use their TTSS to inhibit MAPK activation [34, 35, 42, 43], others activate them. For example, the inflammatory responses induced by the TTSS effectors of Salmonella typhimurium are related to activation of all MAPK, especially p38 which induces IL-8 secretion from epithelial cells [39], and Burkholderia pseudomallei utilizes its TTSS to induce IL-8 secretion and to increase bacterial internalization via activation of p38 and JNK in epithelial cells [44]. Several Vibrio spp. manipulate MAPK signalling pathways to induce Cetuximab ic50 host cell death or disturb the host response to infection [40, 45–49].

Vibrio vulnificus triggers phosphorylation of p38 and ERK via Reactive Oxygen Species in peripheral blood mononuclear cells thereby inducing host cell death [46]. The CtxB cholera toxin from Vibrio cholerae down-regulates p38 and JNK activation in macrophages leading to suppression of production of TNFα and other pro-inflammatory cytokines [40, 47]. Additionally Flagellin A from V. cholerae contributes to IL-8 secretion from epithelial cells through TLR5 and activation of p38, ERK and JNK [48]. Despite the fact that V. parahaemolyticus possesses flagellin proteins similar to those of V. cholerae [49], cells co-incubated with heat-killed V. parahaemolyticus did not exhibit MAPK phosphorylation (Figure 1), suggesting an absence of TLR5 recognition of flagellin.

J Mater Chem 2008, 18:615–620.CrossRef 2. Zhi Ping X, GQ Max L: Layered double hydroxide nanomaterials as potential cellular drug delivery agents. Pure Appl Chem 2006,78(9):1771–1779. 3. Poewe W, Antonini W, Zijlmans JC, Burkhard PR, Vingerhoets F: Levodopa in the treatment of Parkinson’s disease:

an old drug still going strong. Clin Interv Aging 2010, 5:229–238. 4. Aminu Umar K, Samer Hasan Hussein Al A, Mohd Zobir H, Sharida F, Palanisamy A: Development of a controlled-release anti-parkinsonian nanodelivery system using levodopa as the active agent. Int J Nanomedicine 2013, 8:1103–1110. 5. Aminu Umar K, Samer Hasan H-A-A, Mohd Zobir H, Sharida F: Preparation of Tween 80-Zn/Al-levodopa-layered double hydroxides nanocomposite for drug delivery system. Sci World J 2014, 10. Article Selleck CX 5461 ID 104246 6. Suna W, Xiea C, Huafang Wang YH: Specific role of polysorbate 80 coating on the targeting of nanoparticles to the brain. Biomaterials 2004, 25:3065–3071.CrossRef 7. Debanjan D, Senshang L: Double-coated poly (butylcynanoacrylate) nanoparticulate delivery systems for brain targeting of dalargin via oral administration. J Pharm Sci 2005,94(6):1343–1353.CrossRef

8. OECD: OECD guidelines for testing of chemicals. No 407: repeated dose 28-day oral toxicity study in rodents. Paris: Organisation for Economic Co-operation and Development; 2008.CrossRef 9. Redfern WS, Ewart LC, Pierre L, Mark P, Sally R, Jean-Pierre V: Functional assessments in repeat-dose toxicity studies: the art of the possible. Toxicol Res 2013, 2:209–234.CrossRef 10. Prasad Selleck RAD001 SPTLC1 AS: Zinc in human health: effect of zinc on immune cells. Mol Med 2008,14(5–6):353–357. 11. Dandekar P, Dhumal R, Jain R, Tiwari D, Vanage G, Patravale

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Hence, in order to overexpress and purify pneumococcal SmpB, its coding region was cloned in fusion with pneumococcal ssrA (the gene encoding tmRNA) to allow co-expression of both. smpB was amplified by PCR with primers rnm010 and rnm011, which contains a 3’ extension complementary to the 5’-end of ssrA. ssrA was amplified using the primer pair smd057/smd058. The two PCR fragments were then mixed and used as template in a PCR with primers selleckchem rnm010 and smd058. The resulting PCR product was digested with NdeI and BamHI (Fermentas), and cloned into the pET-15b vector (Novagen) previously cleaved with the same restriction enzymes. This construction, named pSDA-02, was first obtained in E. coli DH5α and

then transferred to E. coli BL21(DE3) to allow the expression of His-SmpB. This construct was confirmed by DNA sequencing. Overexpression and purification of proteins RNase R from S. pneumoniae was purified as previously described [30]. For purification of SmpB, BL21(DE3) cells containing pSDA-02

plasmid were grown at 37°C in 250 ml of LB medium supplemented with 100 μg/ml Amp to an OD600 of 0.5. Overexpression Fulvestrant concentration of SmpB was then induced by addition of 1 mM IPTG; induction proceeded for 3 hours at 37°C. Cells were harvested by centrifugation and stored at −80°C. Purification was performed by histidine affinity chromatography using HisTrap Chelating HP columns (GE Healthcare) and AKTA HPLC system (GE Healthcare) as follows. Frozen cells were thawed and resuspended in lysis buffer (50 mM HEPES pH 7.5, 1 M NH4Cl, 5 mM MgCl2, 2 mM β-mercaptoethanol, 10 mM imidazole). Cell suspensions were lysed using a French Press at 9000 psi in the presence of 1 mM PMSF. The crude extracts were treated with Benzonase (Sigma) to degrade the nucleic acids and clarified by a 30 min centrifugation

at 10000 xg. The clarified extracts were then loaded onto a HisTrap Chelating Sepharose 1 ml Histone demethylase column equilibrated with buffer A (20 mM sodium phosphate pH 7.4, 0,5 M NaCl, 20 mM imidazole). Protein elution was achieved by a continuous imidazole gradient (from 20 mM to 500 mM) in buffer A. The fractions containing the purified protein were pooled together and concentrated by centrifugation at 4°C in an Amicon Ultra Centrifugal Filter Device with a molecular mass cutoff of 10 kDa (Millipore). Protein concentration was determined using the Bradford method [59]. SmpB and RNase R purified proteins were loaded in a SDS-PAGE gel and Coomassie blue stained for band excision (data not shown). Bands corresponding to a total of 500 μg of each protein were used to raise antibodies against the respective pneumococcal proteins (Eurogentec). RNA extraction and northern blotting Overnight cultures of S. pneumoniae TIGR4 wild type and mutant derivatives were diluted in pre-warmed THY to a final OD600 of 0.1, and incubated at 37°C until OD600 ~ 0.3. At this point, cultures were split in two aliquots and each aliquot was further incubated at 15°C or 37°C for 2 h.

In the COLD condition body mass decreased from 85.2 ± 11.6 kg to 84.8 ± 11.3 kg and in the RT condition body mass decreased from 85.2 ± 11.6 kg to 84.7 ± 11.3 kg. There were no significant interactions (p=0.223) and subjects did not behave differently to the conditions at the time points. Additionally, there were no significant differences in hydration status between the COLD and RT conditions at either the baseline

time point (p=0.549) or the post-exercise time point (p=0.368). Since atmospheric conditions were also held constant, the authors can Cilomilast datasheet infer that water intake was not different between COLD and RT conditions. Both groups experienced a significant decrease in hydration status from beginning of the exercise session to post-exercise session in both conditions (p<0.0001). There were also no significant interactions between groups and the pre to post-training time point (p=0.209) Table 3. Table 3 Hydration status during training   COLD RT Test PREa POSTa PREa POSTa Weight (kg) 85.2±11.6 84.8±11.3 85.2±11.6 84.7±11.3 Hydration Status (urine specific gravity) 1.00615±0.005 1.01021±0.005 b 1.00564±0.004 1.011942±0.013b aValues represent mean ± standard deviation. bp < .05. Criterion for statistical significance was set at p<0.05. When investigating measures of performance, there were no significant

differences between COLD and RT groups for two of the three performance tests: broad jump (p=0.465) or TTE (p=0.735). Subjects performed broad jumps of 2.17 ± 0.27 m and 2.15 ± 0.25 m in the COLD and RT conditions, respectively. TTE was 638 ± 187 seconds and 643 Pexidartinib ± 189 seconds for the COLD and RT conditions, respectively. However, even though there was not a significant improvement demonstrated, 49% of the participants improved in the broad jump and 51% in the TTE respectively during the COLD. Subjects participating in the RT condition were able to perform significantly more bench press repetitions to failure than when they participated in the COLD condition (p=0.046). During

the COLD condition 22 ± 3.5 repetitions (range: 15–30) were performed and during the RT condition 22.7 ± 3.2 repetitions (range: 17–31) were performed. This was a small, albeit significant, improvement; however, a calculation Fludarabine of an effect size [11] indicates that this would be a negligible to small effect (d=0.2) Table 4. Table 4 Summary of performance test results Test COLDa RTa % of subjects who had improved performance during COLD Bench (reps) 22 ± 3.5 22.73 ± 3.2b 14% Broad Jump (m) 2.17 ± 0.27 2.15 ± 0.25 49% TTE (seconds) 638 ± 187 643 ± 189 51% aValues represent mean ± standard deviation. bp< 0.05. Criterion for statistical significance was set at p<0.05. Discussion Current literature reports that a rise in core temperature can significantly impede performance [1].

The peak positions of χ norm suggest that magnetization reversal mechanism I is predominant for α = 0° and becomes less dominant with increasing α, while the dominance of mechanism II increases with increasing α. Therefore, the maximum in H C for α = 60° and α = 75° could be understood as the result KU 57788 of an interplay between the two magnetization reversal modes. The exact type of these magnetization reversal mechanisms could not be identified by the conducted hysteresis loop measurements. Nevertheless,

one might speculate that these reversal modes are most probably the transversal and vortex magnetization reversal mode as found by micromagnetic simulations for Ni nanowires Erlotinib mouse by Han et al. [25]. Correlating these magnetic results with the structural characterization, one could understand the comparatively high coercivity of the Co nanowires as a direct consequence of the small grain size accompanied by the high amount of grain boundaries that hinder the domain wall movement. The small grain size

itself is most probably a consequence of the deposition via the two simultaneously occurring Co deposition processes, as already discussed in the first part of this paper. Conclusions The electrochemical growth of Co nanowires in ultra-high aspect ratio InP membranes could be successfully characterized by the analysis of the FFT-IS data. The corresponding fit model is represented by a rather complex Abiraterone clinical trial electric equivalent circuit containing a series

resistance and three RC elements. This fit model is not limited to the Co deposition but has also been successfully applied for the deposition of Ni in ultra-high aspect ratio InP membranes. Based on the impedance data, the Co nanowire growth could be divided into two separate processes, most possibly the direct Co deposition and the indirect Co deposition via Co(OH)2. The share of each Co deposition process on the overall Co deposition can be determined directly from the transfer resistances of the two processes obtained from the fitted impedance data. These also indicate a beneficial effect of boric acid on the Co deposition. This characterization of the Co deposition process by FFT-IS will help in optimizing the deposition parameters such as temperature, deposition current, electrolyte composition, etc. with respect to the crystal orientation and thus also of the magnetic properties necessary for the application in magnetoelectric 1– 3 composites. Acknowledgements This work was funded by the DFG as part of the special research field 855 ‘Magneto-electric composite materials – biomagnetic interfaces of the future.’ References 1. Wakai RT, Leuthold AC, Martin CB: Atrial and ventricular fetal heart rate patterns in isolated congenital complete heart block detected by magnetocardiography. Am J Obstet Gynecol 1998,179(1):258. 10.1016/S0002-9378(98)70282-0CrossRef 2.

Efforts to determine the effect of the infection with H. pylori rocF- strains in the cellular infiltration of the gastric mucosa are currently underway. To the JAK inhibitor best of our knowledge, there is only one published work trying to measure the levels of H. pylori arginase in gastric biopsies of patients with gastritis and its correlation with disease [34]. That work showed that there is a lot of variability on the levels of H. pylori arginase in biopsies

but the authors were not able to establish a correlation with the degree of gastritis. The reason for the increased number of genes modulated by the rocF- H. pylori, when compared to the WT and the rocF + bacteria, is not known; however, our results, rather than suggesting the existence of H. pylori arginase mutants in human gastric lesions, highlights the importance that this enzyme may have in the interaction of the bacteria Bcl-2 inhibitor with cells in the human gastric mucosa, and through them, with the immune system. Taken together our results suggest that H. pylori arginase, by modulating the production of IL-8 may play a significant role in the survival of H. pylori in the gastric environment. By preventing an over-zealous immune response, H. pylori can achieve its chronicity through the production of arginase and probably other bacterial factors that contribute to the overall global success of this important and highly-adapted

gastric human pathogen. Conclusion Our results highlight the importance of H. pylori arginase in the modulation of inflammatory responses. Since IL-8 is pivotal for the infiltration of inflammatory cells into the gastric mucosa, H. pylori arginase may be involved in reducing the tissue damage associated with the evolution of the gastric lesions through the modulation of multiple pathways on the gastric epithelial cells. Methods Bacterial growth conditions H. pylori 26695 strains (wild type [WT], rocF- mutant, and the rocF- mutant chromosomally-complemented with wild type 26695 rocF- (rocF-26695-MLB0004, hereafter referred to as rocF+) were described previously [13]. All strains were passaged every 2–3 days on Campylobacter blood buy Rucaparib agar (CBA) plates at 37°C with 85% N2,

5% O2, and 10% CO2. Prior to coculture experiments, H. pylori cells were grown in Ham’s F-12 with heat-inactivated 1% (v/v) fetal bovine serum (FBS) [35]. H. pylori growth was monitored by ATP level using Cell Titer-Glo® cell viability assay kit (Promega, NY, USA), as validated previously [36] and by plating for colony forming units. Comparable number of viable bacteria was assured in each experiment. Tissue culture and co-culture AGS gastric epithelial cells (ATCC CRL-1739, Rockville, MD) were maintained in F-12 with heat-inactivated 10% FBS at 37°C in an atmosphere of 5% CO2. For the experiments, 1 x 106 AGS cells were seeded into 6-well plates containing 2 ml fresh F-12 supplemented with 3% heat-inactivated FBS and cultured for 8 hours.