Genomic DNA was isolated from R16-18d and the sequence of the 16s rRNA gene was determined to be identical to the sequence from NCTC 8325-4 and RRSA16 as described above.

MICs were determined on microdilution plates according to Wiegand et al. (2008) using CAMHB2 as the growth media. Sodium chloride was added to a final concentration of 2% (w/v) when oxacillin was tested. BSA (0.02% w/v) was added to media when vancomycin, ramoplanin or nisin was tested to prevent peptide adhesion to polystyrene. Doubling times were calculated as described (Cui et al., 2003), with tryptic soy broth (TSB) cultures growing Selleck CH5424802 at 37 °C with aeration in the exponential phase [Eqn. (1)], where t1 and t2 are the times of measurement: (1)

Staphylococcus aureus cultures were grown in TSB supplemented with 0.02% BSA (TSB+BSA) at 37 °C with shaking at 200 r.p.m. to OD620 nm≈0.4 and were then treated with an antibiotic. The cultures were then incubated at 37 °C with shaking at 200 r.p.m. Samples were removed periodically for OD measurements and viable counting. Staphylococcus aureus cultures were grown in TSB+BSA at 37 °C with aeration to an OD620 nm of ≈0.7. Samples were removed, pelleted and resuspended in 4% glutaraldehyde. The pellets were washed twice in 0.1 M sodium cacodylate buffer containing 7.5% sucrose and pre-embedded in 1% agar. The samples were washed twice with check details 0.1 M sodium cacodylate buffer containing 7.5% sucrose and postfixed in 1.0% osmium

tetroxide Vorinostat chemical structure in 0.15 M sodium cacodylate buffer. Samples were washed for 10 min twice in 0.11 M veronal acetate buffer. Samples were then dehydrated in an ascending ethanol series and embedded in Epon resin. Sections were cut at 80 nm on a Reichert Ultracut S ultramicrotome and mounted on copper rhodium 200 mesh 3 mm grids. Samples were stained with uranyl acetate for 30 min, rinsed three times in distilled water, stained with Reynold’s lead citrate stain prepared as described by Venable & Coggeshall (1965) for 5 min and rinsed three times in distilled water. Samples were viewed using a Philips/FEI CM12 transmission electron microscope at 80 kV. Cell wall thickness was calculated as described elsewhere (Cui et al., 2000). Twenty radial lines arranged regularly at angles of 18° were placed over the center of images of equatorially cut cells at a final magnification of × 35 000 and the thickness of the cell wall was measured from at least 10 different points. The thickness of the cell walls of 20 cells from each strain was measured. Results are reported as means±SD. The diameter of the 20 cells from each strain was also measured using 20 radial lines arranged regularly at angles of 18° and placed over the center of equatorially cut cells; the results were reported as means±SD. The statistical significance of the data was evaluated using a Student’s t-test.

Furthermore, antiviral treatment, which has led to a clinical improvement, has been shown to reduce HHV8 viral load in patients with KS [63], PEL and haemophagocytic syndrome [64]. In a series of three patients treated with ganciclovir, there was

a reduction in the frequency of acute symptoms of MCD for two patients treated with oral and intravenous ganciclovir [65]. For the third patient, there was resolution of pulmonary and renal failure with intravenous ganciclovir. All the patients had a reduction in HHV8 viral load with the ganciclovir therapy, accompanying the resolution of their symptoms. However, the use of foscarnet and cidofovir antiviral therapy was ineffective in an HIV-negative MCD patient with proven HHV8 viraemia and treatment with corticosteroids in combination with FDA approved Drug Library supplier chlorambucil

chemotherapy was required to achieve a clinical response [66]. Furthermore, the HHV8 viral load rose in this patient with the commencement of anti-herpesvirus therapy; this may indicate that the antiviral therapy was ineffective in this case, or that, once the MCD is established, HHV8 has a less prominent role and antiviral therapy is less this website effective than immunotherapy or chemotherapy. Casper et al. [36] randomized 26 men with HHV8 infection to receive 8 weeks of valganciclovir administered orally (900 mg once per day) or 8 weeks cAMP of placebo. After a 2-week washout period, participants in each group received the study drug they had not yet taken (either valganciclovir or placebo), for 8 additional

weeks. Oral swab samples were collected daily during the study, and HHV8 and CMV DNA were quantified by real-time PCR. A total of 16 HIV-positive men and 10 HIV-negative men enrolled in, and completed the study. Of the 3439 swab samples that participants had been expected to provide, 3029 (88%) were available for analysis. HHV8 was detected on 44% of swabs collected from participants who were receiving placebo, compared with 23% of swabs collected from participants who were receiving valganciclovir (relative risk [RR] 0.54, 95% CI: 0.33–0.90; p = 0.02). Valganciclovir reduced oropharyngeal shedding of cytomegalovirus by 80% (RR: 0.20, 95% CI: 0.08–0.48; p < 0.001). Shedding of HHV8 and shedding of cytomegalovirus were independent. Haematological, renal, or hepatic toxicities were no more common among participants who received the active drug, compared with those who received placebo, though participants who received valganciclovir reported more days of diarrhoea. Valganciclovir administered orally once per day is well tolerated and significantly reduces the frequency and quantity of HHV8 replication. A further study [67] compared the efficacy of valaciclovir, famciclovir and cART in reducing HHV8 oropharyngeal shedding in 6036 swabs from 58 participants.

Mutants H213A and D228A were obtained similarly by using the pair of primers NopT1-H213A-F/NopT1-H213A-R and NopT1-D228A-F/NopT1-D228A-R, which simultaneously introduced an EaeI and a PvuI restriction site, respectively. Mutants nopT1-DKM and nopT1-GCC were obtained by PCR amplification as described earlier using the pair of primers NopT1-DKM-F/NopT1-DKM-R and

NopT1-GCC-F/NopT1-GCC-R, respectively. The primers were designed to obtain the D47A, K48A, and M49A substitutions in case of the NopT1-DKM mutant and G50A, C52S, and C53S substitutions in case of the NopT1-GCC mutant. All mutations were confirmed by diagnostic restriction digestions taking advantage of SacII and NheI sites designed in the primers and sequencing. C-terminally polyhistidine-tagged wild-type NopT1 and NopT2, as well as mutant derivatives of NopT1, were obtained by cloning the respective

coding regions without the stop codons following PCR amplification from the pT7-7 expression Avasimibe price constructs with the pair of primers NopT1-F1/NopT1-R3 and NopT2-F1/NopT2-R3, respectively. The amplicons were digested with appropriate restriction enzymes selleck compound and subcloned into the pET26b vector (Novagen), ligated, and transformed into E. coli strain BL21 (DE3). For protein expression, E. coli BL21 (DE3) transformants harboring the pET26b constructs were grown in LB medium to an OD600 nm of 0.6 at 37 °C, and protein expression was induced for 4 h at 30 °C by adding 0.5 mM isopropyl β-d-thiogalactopyranoside (IPTG). Bacterial cells were collected by centrifugation, old resuspended in lysis buffer (50 mM NaH2PO4, pH 8.0, 300 mM NaCl, 10 mM imidazole) supplemented with 1 mM phenylmethylsulfonyl fluoride (PMSF), and lysed by the addition of lysozyme followed by sonication. Histidine-tagged wild-type and mutant proteins were expressed in E. coli BL21 (DE3) at 30 °C and purified by Ni2+-NTA affinity chromatography under native conditions according to the standard protocol (Qiagen). Proteins were resolved in 14% SDS-polyacrylamide gel electrophoresis (PAGE) and were visualized by Coomassie blue staining and immunoblotting using alkaline phosphatase (AP)-conjugated

anti-His antibody (Qiagen). Protein concentrations were estimated by Coomassie blue staining of SDS-PAGE gels using BSA standards. Prestained molecular size standards (Broad range; New England Bio-Labs) were used to estimate the molecular mass of proteins. Proteins were purified under nondenaturing conditions as mentioned earlier and lyophilized, and their protease activity was determined using resorufin-labeled casein (Roche) as a substrate. Lyophilized samples were dissolved in different buffers at pH range 5.5–9.5 in final volume of 100 μL and preincubated at 37 °C for 1 h. The enzymatic activity was determined in 50 mM buffers (sodium acetate buffer at pH 5.5; potassium phosphate at pH range 6.5–7.5; Tris at pH range 8.5–9.5) containing 10 mM l-cysteine, 10 mM EDTA, and 0.4% casein in a final volume of 200 μL.

, 1999). Macrophages from wild-type mice are more effective at inhibiting S. Typhimurium replication than mCRAMP−/− macrophages (Rosenberger et al., 2004). Together, these experiments

indicate that defensins and cathelicidins are important in the host defense against S. Typhimurium infection. Conversely, in a study of S. Typhimurium mutants selected for sensitivity to AMP-mediated killing, eleven out of twelve AMP-sensitive bacterial strains displayed decreased virulence in a mouse infection model, indicating that AMP resistance may be a critical co-requisite for bacterial virulence (Groisman et al., 1992). Animal models have provided evidence for the role of AMPs in other GSK126 in vivo Gram-negative bacterial infections as well. mCRAMP−/− mice are more susceptible to intestinal infection with Citrobacter rodentium (Iimura et al., 2005) and urinary tract infection with UPEC (Chromek et al., 2006). Newborn rats treated

with a chemical that damages AMP-producing Paneth cells become more susceptible to infection with enteroinvasive E. coli (EIEC) (Sherman et al., 2005). Conversely, treatment of Shigella-infected rabbits with butyrate led to upregulation of cathelicidin and marked clinical improvement and survival rates (Raqib et al., 2006), and in a human xenograft model, LL-37 overexpression increased www.selleckchem.com/products/apo866-fk866.html killing of Pseudomonas aeruginosa (Bals et al., 1999). AMPs are important to control colonization by not only bacterial pathogens but

also commensal bacteria. A recent study revealed that aberrant expression of Paneth cells α-defensins alters the composition of the intestinal microbiota without changing the total bacterial numbers (Salzman et al., 2010). This finding raises the possibility that differences in pathogen susceptibility described for animals with aberrant AMP expression or activity may, in Sodium butyrate part, be mediated indirectly by changes in the microbiota. To survive the bactericidal action of AMPs, bacteria must sense the presence of AMPs and adapt accordingly by precisely controlling the expression of genes involved in AMP resistance. In Enterobacteriaceae, genes controlling AMP resistance are usually under the control of the two-component signaling pathways PhoPQ and PmrAB and the RcsBCD phosphorelay system. In S. Typhimurium, PhoPQ controls PmrAB signaling by promoting the expression of the PmrD protein that binds to phosphorylated PmrA and prevents dephosphorylation, resulting in sustained activation of PmrA-regulated genes (Bijlsma & Groisman, 2003). There is compelling evidence that AMPs are sensed directly by the PhoQ sensor kinase. Following self-promoted uptake through the OM, α-helical AMPs such as LL-37 and C18G bind directly to an anionic region of the PhoQ periplasmic domain and activate the PhoPQ system, leading to expression of PhoP-activated genes (Bader et al., 2005).

, 2003). Ftn and Bfr function similarly as iron-storage proteins, preserving iron in a nonreactive form that can be released and used

as a nutrient source during conditions of iron starvation (Abdul-Tehrani et al., 1999; Chen et al., 2010). Dps proteins are involved in iron detoxification. Dps proteins protect DNA from the harmful Fenton reaction by catalysing the oxidation of two ferrous iron molecules for every one hydrogen peroxide (H2O2) molecule and thus prevent the production of toxic hydroxyl radicals (Zhao et al., 2002; Ceci et al., 2003). The erythrin-vacuolar iron transport (Er-VIT1) http://www.selleckchem.com/products/LDE225(NVP-LDE225).html protein, a member of the Ferritin-like superfamily, has a distinct structure consisting of two major domains (Fig. 1) (Andrews, 2010). First, the N-terminal Er or Ferritin-like domain contains the four-helical bundle and conserved amino acid residues for a di-iron site. Second, the C-terminal domain is a membrane-embedded VIT1 domain that is homologous to Arabidopsis VIT1, which is involved in iron transport into vacuoles (Kim et al., 2006). Arabidopsis VIT1 has a 62% http://www.selleckchem.com/products/r428.html amino acid similar to the yeast Ca2+-sensitive cross-complementer 1 (CCC1) protein. CCC1 is an iron/manganese transporter that transfers iron from the cytoplasm to vacuoles (Li et al., 2001). At present, the Er-VIT1 protein has not been characterized, and thus, the protein’s function

is still not known. The A. tumefaciens mbfA gene (Atu0251), a member of Er-VIT1 family, encodes a putative membrane-bound ferritin (MbfA) that is predicted

to be regulated by the iron response regulator (irr) (Rodionov et al., 2006). In closely related Rhizobium leguminosarum and Bradyrhizobium japonicum bacteria, it has been demonstrated that transcription of mbfA is regulated by Irr in response to iron (Rudolph et al., 2006; Todd et al., 2006). Agrobacterium tumefaciens Irr co-modulates iron homeostasis with the rhizobial iron regulator (RirA), in which Irr plays a contrasting role in positively controlling iron uptake and transport genes (Hibbing & Fuqua, 2011). However, the regulation and physiological function of A. tumefaciens mbfA have not been studied. Here, an A. tumefaciens mbfA mutant strain was generated to investigate the physiological functions of Bcl-w mbfA in response to iron and H2O2 stresses. Agrobacterium tumefaciens strains used in this study include the wild-type strain (NTL4), a Ti plasmid-cured derivative of strain C58 (Luo et al., 2001), a catalase-deficient strain (KC05, katA and catE double mutation) (Prapagdee et al., 2004) and a rhizobial iron regulator mutant strain (PN094, previously named NTLrirA) (Ngok-ngam et al., 2009). Agrobacterium tumefaciens strains were grown aerobically at 28 °C in Luria–Bertani (LB) medium or on LB plates containing 1.5% agar (LA), supplemented with 100 μg mL−1 carbenicillin (Cb), 25 μg mL−1 chloramphenicol (Cm), 90 μg mL−1 gentamicin (Gm) or 30 μg mL−1 kanamycin (Km), as required. Escherichia coli strains BW20767 (Metcalf et al.

5 nm with 1-nm bandwidth at a scan speed of 50 nm min−1. Averages of five scans were obtained for blank and protein spectra, and data were corrected for buffer contribution. Measurement was taken at protein concentration between 1 and 2 μM under nitrogen flow. The results are expressed

as mean residue ellipticity in units of degree cm−2 dmol−1. Xenocin is a multi-domain toxic protein consisting of translocation domain, receptor domain and catalytic domain. Toxicity of xenocin lies in its catalytic domain. To study the detrimental selleck chemical effect of xenocin alone, it was cloned under tightly regulated ara promoter. Xenorhabdus nematophila was not able to uptake arabinose, which is inducer for ara promoter. Therefore, all the endogenous toxicity assays were performed in the E. coli TOP10, the recommended host for the expression vector containing ara promoter like pBAD. In the endogenous toxic assay, growth profile of arabinose-induced JSR4 strain containing vector alone was considered as 100% and compared with induced JSR2 strain containing xenocin alone. Results showed that there was no change in growth profile of JSR2 strain after first hour of induction; however, growth was inhibited by 50% after second hour and was further Dorsomorphin mouse declined in consecutive hours as

shown in Fig. 1. In case of catalytic domain, growth declined immediately after induction and it was inhibited by almost 70% in first hours of induction, 80% in second hour and was further declined in the consecutive hours NADPH-cytochrome-c2 reductase as shown in Fig. 1. In our previous work, we have shown that catalytic domain of xenocin has RNase activity (Singh & Banerjee, 2008). On the basis of multiple sequence alignment (Supporting Information, Fig. S1) and homology model, six conserved amino acids residues were predicted to form active site in catalytic domain including D535, H538, E542, H551, K564 and R570 as shown in Fig. 2a. Catalytic mechanism of RNA hydrolysis has been thoroughly

studied by protein engineering and crystallography (Gilliland, 1997). RNase A has two active histidine residues that cooperate during the catalytic cycle (Raines, 1998; Scheraga et al., 2001). Other ribonuclease, such as barnase and colicin E3, precede probably through the similar mechanism, but in these cases, histidine and glutamic acid act as catalytic residues (Walker et al., 2004) Figs. S2, S3, S4 and S5. Killing of the target cells by multi-domain E colicins occur in three different stages. First step to bind with receptor, followed by its translocation into the periplasmic space and finally endogenous toxicity in the cytoplasm of target cells by its catalytic domain (Carr et al., 2000). Primary sequence of catalytic domain from xenocin revealed the presence of four histidine residues. Interestingly, three of them were found conserved in multiple sequence alignment (Fig. S1).

5 nm with 1-nm bandwidth at a scan speed of 50 nm min−1. Averages of five scans were obtained for blank and protein spectra, and data were corrected for buffer contribution. Measurement was taken at protein concentration between 1 and 2 μM under nitrogen flow. The results are expressed

as mean residue ellipticity in units of degree cm−2 dmol−1. Xenocin is a multi-domain toxic protein consisting of translocation domain, receptor domain and catalytic domain. Toxicity of xenocin lies in its catalytic domain. To study the detrimental GSK J4 effect of xenocin alone, it was cloned under tightly regulated ara promoter. Xenorhabdus nematophila was not able to uptake arabinose, which is inducer for ara promoter. Therefore, all the endogenous toxicity assays were performed in the E. coli TOP10, the recommended host for the expression vector containing ara promoter like pBAD. In the endogenous toxic assay, growth profile of arabinose-induced JSR4 strain containing vector alone was considered as 100% and compared with induced JSR2 strain containing xenocin alone. Results showed that there was no change in growth profile of JSR2 strain after first hour of induction; however, growth was inhibited by 50% after second hour and was further LY294002 datasheet declined in consecutive hours as

shown in Fig. 1. In case of catalytic domain, growth declined immediately after induction and it was inhibited by almost 70% in first hours of induction, 80% in second hour and was further declined in the consecutive hours Alectinib as shown in Fig. 1. In our previous work, we have shown that catalytic domain of xenocin has RNase activity (Singh & Banerjee, 2008). On the basis of multiple sequence alignment (Supporting Information, Fig. S1) and homology model, six conserved amino acids residues were predicted to form active site in catalytic domain including D535, H538, E542, H551, K564 and R570 as shown in Fig. 2a. Catalytic mechanism of RNA hydrolysis has been thoroughly

studied by protein engineering and crystallography (Gilliland, 1997). RNase A has two active histidine residues that cooperate during the catalytic cycle (Raines, 1998; Scheraga et al., 2001). Other ribonuclease, such as barnase and colicin E3, precede probably through the similar mechanism, but in these cases, histidine and glutamic acid act as catalytic residues (Walker et al., 2004) Figs. S2, S3, S4 and S5. Killing of the target cells by multi-domain E colicins occur in three different stages. First step to bind with receptor, followed by its translocation into the periplasmic space and finally endogenous toxicity in the cytoplasm of target cells by its catalytic domain (Carr et al., 2000). Primary sequence of catalytic domain from xenocin revealed the presence of four histidine residues. Interestingly, three of them were found conserved in multiple sequence alignment (Fig. S1).

Also, the statistical model does not use the mean values for each subject but takes all valid observations into account. In the control group, the mean latency of voluntary saccades in No-discrimination/No-change trials was 391 ms [364, 417], the intercept of the model. In this baseline condition, the PD group made saccades at latencies that were 71 ms [32, 110] longer than in the control group (t38 = 3.69, P < 0.001). In the control group

in No-discrimination trials, the peripheral symbol-changes did not significantly affect saccade latencies: there was a small latency increase of 10 ms [−13, 33] (t38 = 0.85, P = 0.40). In contrast, in the PD group in No-discrimination trials, the symbol-changes reduced latencies by 26 ms [2, 49] (t38 = –2.23, P = 0.03) compared with No-change trials. The discrimination task reduced latencies in the control group,

by find more 33 ms [9, 58] (t76 = –2.70, P = 0.01). In the PD group, the effect of the discrimination task on latencies was significantly larger, with latencies reduced by an additional 37 ms [2, 71] over and above the 33 ms reduction in the control group (t76 = –2.09, P = 0.04). In discrimination trials, the symbol-changes no longer abnormally affected saccade latencies in the PD group. Figure 2 shows the uncorrected mean group latencies [95% CI] calculated from each participant’s mean latency in each of the four trial types, No-discrimination/No-change, No-discrimination/Change, Discrimination/No-change and Discrimination/Change trials. The mean primary gain of GDC-0068 chemical structure voluntary saccades in No-change trials without the discrimination task in the control group was 0.85 [0.82, 0.89], the intercept of the model. In this baseline condition, the PD group’s primary gain was 0.06 [0.01, 0.11] smaller (t38 = –2.42, P = 0.02). The discrimination task increased gain values in both groups: in the control group the discrimination task increased gain by 0.05 [0.02,

0.08] (t38 = 3.10, P-type ATPase P = 0.01) and in the PD group by 0.04 [0.01, 0.08] (t38 = 2.51, P = 0.02). Gain values were not affected by peripheral symbol-changes. In Distractor and Target/Distractor trials the peripheral symbol changes could potentially interfere with saccade plans as they occurred away from the target location. To assess the effect of the peripheral symbol changes on the production of direction errors (saccades that were not directed at the cued target location) these trials with a symbol-change at a non-target location were pooled into a condition labelled ‘with distractors’. In Target and No-change trials, the symbol-changes were not expected to interfere with saccade plans as they occurred at the target location or not at all. Therefore, No-change and Target trials were combined into a condition labelled ‘without distractors’. There was a significant interaction between the effects of the discrimination task and the distractors (z = −2.82, P = 0.005).

The scenario-based responses suggested a provider tendency toward nonantibiotic therapy and fluid hydration when treating mild to moderate diarrhea. Six to sixteen percent of providers in these scenarios also felt that IV fluids were appropriate stand alone therapy. Furthermore, 64% of providers chose not to use antibiotics for moderate to severe TD while 19% felt that fluids only were sufficient to treat severe inflammatory diarrhea. These prescribing behaviors generally go against current practices for these clinical-based scenarios.6,8,17,18

In all of the scenarios a low percentage of providers prescribed combination therapy of antimotility agents with antibiotics, a strategy which has been found to significantly reduce the duration of illness compared to antibiotics alone Y-27632 clinical trial in IBET762 most cases of uncomplicated watery diarrhea.13 Of particular concern, the current study finds that many of the military providers continue to recommend fluids only or antimotility agents for treatment of TD (independent of severity). It may be that providers base these management decisions on treatment of acute gastrointestinal infections

in the United States, which are known to be predominantly viral in origin. Although some resources recommend these agents alone in mild cases of diarrhea, including the revised edition of US Army Center for Health Promotion and Preventive Medicine Technical Guide-273,18 it may be advisable to treat even these mild cases more aggressively depending on the operational tempo given the potential impact on mission readiness and the predisposition to dehydrating comorbid illness in the austere deployment environments. Providers’ responses to amount of time off and limited duty given to soldiers with TD is an important reflection of the burden these common

infections have on the fighting strength. With 46% of providers saying Reverse transcriptase they would sometimes confine those soldiers with diarrhea to quarters, and 14% saying they would often confine to quarters, the amount of duty days lost due to these frequent illnesses are considerable.19 These data are concordant with observations obtained directly from afflicted soldiers where Sanders and colleagues reported that nearly half of troops surveyed who developed diarrhea went to seek medical care at least once, and 46.1% of episodes of diarrhea resulted in decreased job performance.9 The provider attitudes toward antimotility agents revealed some common misunderstandings regarding treatment options for TD. The majority of providers felt that antimotility agents kept toxins or pathogens inside the body and could lead to more intestinal damage. The majority also felt that antimotility agents prolonged illness by delaying excretion of the pathogen.

05 compared with the control media and L. rhamnosus HN001) (Fig. 1b). Lactobacillus

plantarum DSM 2648 also had a similar effect on TEER when tested using differentiated Caco-2 monolayers (18 days old) (Fig. 1c). This study demonstrates the strain-dependent effects of lactobacilli on intestinal barrier function and that all strains of the same species should not be assumed to have similar health-promoting properties. Sorafenib Lactobacillus plantarum are effective in enhancing TEER, with three out of the five L. plantarum isolates tested having a positive effect on TEER compared with the control media. A number of human oral isolates were also effective in enhancing TEER compared with the control media. Three out of four L. rhamnosus isolates, the L. paracasei isolate and the L. oris isolate had a positive effect on TEER.

However, several of the human oral isolates had a negative effect on TEER; three out of five L. fermentum isolates and the L. jensenii isolate induced a decrease in TEER compared with the control media. In contrast, one isolate of L. fermentum induced an increase in TEER compared with the control media. Lactobacillus plantarum DSM 2648 was chosen for further investigation because it had a greater positive effect on TEER compared with the benchmark, L. rhamnosus HN001, over the 12-h test period. Acid and bile tolerance (2 and 4 h) of L. plantarum DSM 2648 was compared with that of L. rhamnosus HN001 (Fig. 2). Both bacterial Forskolin nmr strains were able to tolerate acidic conditions (pH 4 for 4 h) without the loss of cell viability; however, both strains had a reduced viability of 6–7 log units under conditions of pH 2 for 4 h. The viability of L. rhamnosus HN001 decreased by 2 log units in the presence of 0.5% bile and by 5 log units in the presence of

Rebamipide 1% bile, whereas the viability of L. plantarum DSM 2648 only reduced by 2 log units by 1% bile. The ability of L. plantarum DSM 2648 to adhere to intestinal cells (3 and 6 h) was also compared with that of the benchmark strain, L. rhamnosus HN001 (Fig. 3). Lactobacillus plantarum DSM 2648 adhered in higher numbers (10 times more) to both confluent undifferentiated and differentiated Caco-2 cells compared with L. rhamnosus HN001 (P<0.05 at 3 and 6 h). Lactobacillus plantarum DSM 2648 displayed better in vitro tolerance to gastrointestinal conditions compared with L. rhamnosus HN001, which has been detected in human faeces after ingestion (Tannock et al., 2000); thus, it is possible that L. plantarum DSM 2648 may also survive passage through the human gastrointestinal tract. Lactobacillus plantarum DSM 2648 was also able to prevent the deleterious EPEC-induced TEER changes observed when the EPEC strain was incubated alone (Fig. 1d); however, the action of L. plantarum DSM 2648 was transient, lasting for at most 8 h. The action of L. plantarum DSM 2648 on EPEC interactions with Caco-2 cells was further explored using coculture adherence experiments.