It is notable that

the PTS/glycosidase systems seem to be

It is notable that

the PTS/glycosidase systems seem to be present in gut/commensal bacteria and others such as Clostridium difficile that can colonise the gut. Therefore, it would appear that adaptation to the selleck kinase inhibitor intestinal niche seems to be associated with the presence of substantially higher numbers of genes encoding glycosidase enzymes, particularly those involved in the hydrolysis of disaccharides and oligosaccharides of plant origin. Genes for the metabolism of sugars other than lactose are almost entirely absent from the more nutritionally Vorinostat manufacturer fastidious dairy strains. Another interesting observation was that the degree of similarity between the genes/protein sequences from Lb. helveticus DPC4571 and Lb. acidophilus NCFM was generally much higher than between Lb. acidophilus NCFM and any of the other strains. While Lb. acidophilus NCFM and the other gut and multi-environment strains had very similar complements of glycosidase genes, the sequence

similarity was much lower (with the exception of a few Lb. johnsonii genes) than between the NCFM/DPC4571 sequences, even though there were substantial differences in glycosidase gene content between Lb. acidophilus NCFM and Lb. helveticus DPC4571. The loss of a significant number of glycosidase genes together with the high degree of similarity between the remaining genes suggests that Lb. helveticus DPC4571 selleck chemicals has undergone a relatively recent loss of sugar metabolism capacity relative to its divergence from Lb. acidophilus NCFM. Of the sugar metabolism genes analysed, only one (lba_1689) can be used in our barcode as

a gut organism indicator. Bile Salt Hydrolases Intestinal bacteria can experience a wide number of stresses in the intestinal tract including Janus kinase (JAK) those caused by low pH and presence of bile. In this respect, bile salt tolerance is thought to be an important aspect of survival for bacteria which inhabit the intestinal tract. Most intestinal isolates of lactobacilli and some lactobacilli involved in food fermentations exhibit bile salt hydrolase activity [22, 23]. These enzymes catalyze the hydrolysis of conjugated bile acids, which enter the small bowel in bile and are important for the emulsification, digestion and absorption of dietary lipids present in the proximal small bowel [24]. It has been suggested that deconjugation of bile acids is a detoxification method and protects the cells from conjugated bile. Conversely, negative effects of bile salt hydrolase activity have also been reported including cases of contaminated small bowel syndrome, impaired lipid absorption, gallstone formation, and increased risk of colon cancer [25]. In Lactobacillus-free mice, bile salt hydrolase activity was reduced by 87%, revealing that lactobacilli are the main contributors to bile salt hydrolysis [23].

AFM images in Figure 3 indicate three-dimensional topographies of

AFM images in Figure 3 indicate three-dimensional topographies of magnetic fluorescent nanoparticles. It seems that the NPs have some aggregations, which may be due to the polymer matrix on the surface of NPs with too high concentration selleck resulting in NPs becoming sticky and gluey. The particle average size of magnetic nanoparticles is about 100 nm in diameter. Figure 3 AFM images of magnetic nanoparticles. (a) Height image, (b) corresponding phase image, and (c) 3D rendering of AFM images of magnetic nanoparticles in (a). AFM image of the NP-DNA complex is also analyzed in order to investigate

the binding mechanism between NPs and DNA. As shown in Figure 4a,b, it is apparent that several globes are attached to https://www.selleckchem.com/screening/pi3k-signaling-inhibitor-library.html each individual DNA strand and interact with each other. The blue line trace in Figure 4a shows that the radius of the representative globe is about 50.37 nm, which correlates well with the size of spherical NPs. The results indicate formation of the NP-DNA complexes,

which is in agreement with the agarose gel electrophoresis conclusion. The AFM images further proved an attractive interaction between NPs and DNA leading to the formation of NP-DNA complexes. As shown in Figure 4c, the 3D image of Figure 4b indicates that the NP-DNA complex surface is not smooth due to the magnetic nanoparticles attached on the DNA strand surface. Figure 4 AFM images of NP-DNA complex. (a) Height image (below is the corresponding topographic height profile along the blue line), (b) phase image, and (c) 3D rendering of AFM images of NP-DNA complex in (b). The location of NPs in the cells To verify that the NPs can pass the cell membranes, PK-15 cells were treated with membrane-specific red fluorescent dye DiI for 10 min, and then NPs were incubated in the fluorescently labelled cells with magnetic force-induced sedimentation. After treatments, cells were dyed by DiI to show the red cell membrane location. The green fluorescence signal of NPs can be detected inside the cell after an incubation time of 30 min (Figure 5). Figure 5 Fluorescence images of green magnetic nanoparticles in DiI-labelled BCKDHB PK-15 cells and images with greater magnification.

(a to d) Fluorescence images of green magnetic nanoparticles in PK-15 cells labelled with membrane-specific red fluorescent dye DiI. (e to h) Fluorescence images with greater magnification. As shown in Figure 5a,b,c,d, NPs are internalized as intracellular green fluorescent clusters and the cell was clearly outlined with green cluster enrichment in the find more interior. From the images shown in Figure 5e,f,g,h with greater magnification, the location of NPs inside the cell can be observed clearer. In the process of our experiments, we found that NPs binding to cell membranes occur within few minutes under magnetic field. The presence of intracellular green fluorescent clusters was evidenced by treating NPs for 30 min, which colocalize with the membrane-specific probe DiI.

A substantial reduction in both the number and size of inclusions

A substantial reduction in both the number and size of inclusions was seen with chlamydiae harvested from HeLa cells exposed to compound D7

(bottom panels). Similar results were obtained with undiluted chlamydial lysates and with lysates harvested at 84 hpi (data not shown). Discussion Chlamydiae are obligate intracellular pathogens that have a unique biphasic developmental cycle. We have previously shown that C. pneumoniae contains three Ser/Thr protein kinases and that one of these, PknD, is a membrane-associated Integrin inhibitor kinase that phosphorylates CdsD, a structural protein of the type III secretion system [45]. In the selleck present study we have identified a selective inhibitor of PknD and show that this compound blocks phosphorylation of CdsD in vitro, retards the intracellular growth rate and decreases Smoothened Agonist cell line the number of infectious C. pneumoniae produced following infection of HeLa cells. To elucidate the role of PknD in the chlamydial developmental cycle, we screened a small library of known eukaryotic kinase inhibitors in an attempt to identify

a PknD inhibitor. In this study we show that compound D7 is a potent inhibitor of C. pneumoniae PknD activity in vitro. PknD autophosphorylation and subsequent phosphorylation of the substrate CdsD were completely inhibited by compound D7. When added to C. pneumoniae-infected HeLa cells, the 3′ pyridyl oxindole compound retarded chlamydial replication. The restriction of the developmental cycle was not due to the induction of chlamydial persistence as seen with interferon-γ or iron deprivation [34, 38]

since PB were not detected in inclusions when viewed by electron microscopy. Compound D7 also decreased the number of infectious C. pneumoniae upon passage suggesting that the compound interferes with an essential step in C. pneumoniae development. The mechanism of chlamydial growth retardation by compound D7 is unknown but an involvement of host cell JAK3 is unlikely because the expression of JAK3 is restricted to the hematopoietic cell lineage [49–51] and HeLa cells do not express JAK3. The absence of JAK3 in Chlamydia-infected HeLa cells is supported by a recent study that failed to detect the induction or expression of the JAK3 substrate, STAT5, in C. trachomatis-infected HeLa cells [52]. In addition, other potent JAK3 inhibitors (compounds D4, D5 and D6) did not selleck kinase inhibitor interfere with C. pneumoniae growth in HeLa cells. Therefore the mechanism of C. pneumoniae growth retardation in HeLa cells is unlikely due to an effect of compound D7 on JAK3 activity. Our data also rule out an effect of compound D7 on the MEK/ERK signaling pathway required for chlamydial infection and intracellular growth. Activation of the MEK/ERK pathway has been shown to be essential for chlamydial invasion of HeLa cells [43], and sustained activation of Raf-MEK-ERK-cPLA2 is also required for acquisition of glycerophospholipids and growth by C. pneumoniae [48].

Bandyopadhyay and colleagues were able to apply the same reasonin

Bandyopadhyay and colleagues were able to apply the same reasoning and used 2,3-dichloro-5,6-dicyano-p-benzoquinone which is capable of transforming between four different states to mimic natural phenomenon such as diffusion of heat and detection of cancer growth [54]. Pure computation through DNA DNA has also been applied for the development of pure computational methods. While many techniques are available to use DNA for computation, the most widely used technique involves the manipulation of mixtures of DNA on a support. A DNA molecule which encodes all possible solutions to a designed problem is synthesized and attached to this supportive surface. Repeated hybridization cycles and action of exonuclease

enzymes are used to digest, identify, PRIMA-1MET purchase and eliminate non-solution strands of DNA. Upon completion of this step, several polymerase chain reaction (PCR) reactions are used to amplify remaining molecules, most of which are then hybridized to an array of molecules [55]. Recent progress in DNA computation has been remarkable. Although these advances may be far off to be equivalent of the today’s computational capacities of computers, the long-term goal of this research would be DNA computing, overriding everyday computing with great perfection. DNA physical applications The term nanoelectronics refers to the use of 3-Methyladenine chemical structure nanotechnology for the use and development of electrical components and VX-661 datasheet circuits.

Nanoscale electronics have been developed at the molecular level. Such devices are referred to as molecular electronics [56]. Nanoelectronics had been highly dependent on the complementary-symmetry metal-oxide semiconductor (CMOS) technology. CMOS has been vital in analogue circuits such as image sensors, data convertors, and logic-based devices such as digital logic circuits, microcontrollers, and microprocessors [57]. However, CMOS is being replaced as the demand for further Erastin clinical trial miniaturization and processing speeds increase. CMOS circuitry has limitations that can greatly influence the size and shape of computers and other electronics.

DNA offers a solution to these problems. Carbon nanotube devices and wires have been developed through self-guided assembly [58]. These materials are capable of forming electronic devices such as nanowires like those shown in Figure 7 and transistors [59, 60], thus behaving very similarly to a typical CMOS circuit. The advantage of such devices is that DNA can be accumulated in larger densities and numbers as compared to a typical circuit in a normal electrical system. In addition, DNA is fairly efficient in terms of power consumption and cost [58]. Figure 7 DNA uncoiling and forming precise patterns, a prelude to biologically based electronics and medical devices [61]. DNA wires, transistors, capacitors and other devices DNA self-assembly is essential to form any nanoscale biological device. Prior to the development of nanowires, mostly B-DNA was used.

Then, neo2 from pMNMM2 was removed by SalI and SmaI and replaced

Then, neo2 from pMNMM2 was removed by SalI and SmaI and replaced with the amplified neo5 cassette, resulting in pMNMM3 (Fig. 1A). The DNA sequence of pMNMM3 can be found in the Additional file 1. A Cre-recombinase (DDBJ/EMBL/GenBank AAG34515) encoding DNA, which was optimized for Tetrahymena codon-usage, was synthesized (MR. GENE GmbH, Regensburg, Germany) and named cre1. An HA sequence including a short two-amino acid linker LDN-193189 (GA) was added at the N-terminus

of cre1 by PCR amplifying the cre1 coding sequence using PrimeStar HS DNA Polymerase (Takara) with the primers HA-GA-Cre-NdeFW and Cre-MluRV. Then, this PCR product was cloned into NdeI and MluI sites of pMNMM3 to produce pMNMM3-HA-cre1 (Fig. 1B). The MTT1-5′-1-neo5-MTT1-5′-2-HA-cre1-MTT1-3′ construct was excised from the vector backbone by digesting pMNMM3-HA-cre1 with XhoI and SpeI. The DNA sequence of pMNMM3-HA-cre1 can be found in the Additional file 1. Construction of the loxP-neo4-loxP-EGFP-TWI1 construct by PCR First, the loxP-neo4-loxP sequence was generated by PCR amplifying the neo4 cassette with the primers LoxNeoFWXho and LoxNeoRV. These primers had loxP selleck kinase inhibitor sequences at their 5′-termini. PrimeStar HS DNA Polymerase (Takara) was used for all PCR reactions in this section.

In parallel, EGFP was amplified by PCR with the primers LoxGFPFW and LoxGFPRVBam using pOptiGFP as a template. pOptiGFP has a EGFP sequence optimized for Tetrahymena codon-usage (Kataoka et al. submitted with this manuscript). A short complementary CRM1 inhibitor sequence was designed at the 3′-terminus of loxP-neo4-loxP and the 5′-terminus of EGFP. Then, loxP-neo4-loxP and EGFP PCR products were concatenated by overlapping PCR with LoxNeoFWXho and LoxGFPRVBam. The resulting loxP-neo4-loxP-EGFP was cloned into the BamHI and XhoI sites of pBlueScript SK(+) to create ploxP-neo4-loxP-EGFP. The loxP-neo4-loxP-EGFP-TWI1 construct (see Fig. 3A) was generated by PCR. The 5′-flanking

Ponatinib manufacturer and N-terminal regions of the TWI1 gene were amplified using the primers TWI15LoxFW + TWI15LoxRVATGplus and TWI1 NGFPFW + TWI1NGFPRV, respectively, resulting in TWI1-5F and TWI1-N. Also, loxP-neo4-loxP-EGFP was excised from ploxP-neo4-loxP-EGFP using BamHI and XhoI. This fragment had overlapping sequences with the 3′ terminus of TWI1-5F and with the 5′- terminus of TWI1-N, respectively. Finally, the three DNA segments, TWI1-5F, loxP-neo4-loxP-EGFP and TWI1-N were combined by overlapping PCR using TWI15LoxFW and TWI1 NGFPRV. The PCR product loxP-neo4-loxP-EGFP-TWI1 was purified and used directly for the transformation of Tetrahymena. Construction of Tetrahymena strains CRE556 and loxP-neo4-loxP-EGFP-TWI1 Biolistic gun transformation was performed as described [2] to introduce the constructs into the macronucleus by homologous recombination. The B2086 and CU428 wild-type strains were transformed with the digested pMNMM3-HA-cre1 and the loxP-neo4-loxP-EGFP-TWI1 PCR products, respectively.

Side Reach 45–54 93 s 0 22 55–65 0 40 The men with early OA all s

Side Reach 45–54 93 s 0 22 55–65 0 40 The men with early OA all scored above p5, except on the dynamic bending test. One of the older men scored below p5 on the overhead working posture test. On all tests, 20–40% of the younger women and 25–65% of the older women scored below p5. Discussion This study revealed that both the 15 male and the 78 female subjects from a subsample from the CHECK cohort at baseline reported

a worse physical health status (SF-36) compared to the healthy ageing workers, whereas the women also reported a worse mental health status on 3 out of 4 scales. On the FCE, the female CHECK subjects performed significantly lower than their healthy working counterparts on all https://www.selleckchem.com/Proteasome.html 6 tests. The male subjects with OA performed lower on 3 out of 6 tests. A substantial proportion of female subjects demonstrated functional capacities that would be considered insufficient to meet the lowest category of physical job demands. The worse physical health status as reported on the SF-36 can be attributed to the knee or hip complaints of the subjects, but other physical factors may also have influenced their health status. Serious comorbidity was an exclusion criterion for the CHECK cohort, but back pain and other musculoskeletal discomfort were frequently reported. Contrarily, an over representation of physically ITF2357 nmr strong and healthy volunteers in the reference population

may have introduced bias that explains part of the observed GDC0449 differences. Still, the early phase of OA is clearly accompanied by self-reported limitations in physical function and physical roles for both sexes and also by mental health limitations for women. The worse self-reported health status of the subjects with early OA compared to the healthy working subjects was also reflected in a lower functional capacity as measured on the FCE. The pain and stiffness in

the hips or knees, possibly in combination with other health complaints, seem to have affected their performance in work-related physical activities. We reported earlier that in this sample the subjects with low self-reported functional status showed Celecoxib lower performances on the FCE (Bieleman et al. 2009). About half of the subjects with early OA in this study did not have a paid job. Either or not having a paid job has been reported to explain part of the performance on an FCE (Bieleman et al. 2007). For example, on ‘lifting low’ the average difference between women from this study with paid work and those without paid work was 4.7 kg (19.4 kg vs. 14.7 kg). However, after correcting for this factor, there still remains a substantial difference between the capacities of the working subjects with early OA and the reference group of healthy workers. Therefore, it was concluded that in the early phase of OA of the hips and knees a decreased functional capacity is seen, both in working people and even more in people without paid work.

It is generally accepted that activation of Hog1p in the absence

It is generally accepted that activation of Hog1p in the absence of osmotic stress results in growth inhibitory effects [46]. Previously we reported that the antifungal effects of fludioxonil, iprodione and ambruticin VS3 are dependent on the Ssk1 – Pbs2 – Hog1p branch of the osmotic stress response pathway [25], so that a prerequisite for phosphorylation of Hog1p is the non-phosphorylated form of the response regulator Ssk1p [47]. It was even reported that the

presence of phosphorylated LY2603618 manufacturer Ssk1p prevented the activation of the MAP3K Ssk2p from unphosphorylated Ssk1p [48]. Ssk1p receives phosphate groups indirectly from HKs via the histidine transfer protein Ypd1p. Our results indicate that this phosphorylation is inhibited only in strains which are exposed to osmotic

stress or which express the wild-type CaNIK1 variants and are treated with fungicides. In strains expressing mutated non-functional CaNIK1 phosphorylation of Ssk1 was not inhibited. This conclusion is in agreement with [23] who showed that fludioxonil treatment of S. cerevisiae expressing the group III DhNik1p decreased the phosphate transfer to a response regulator even in the presence of the endogenous, active HK Sln1. Group III HKs are characterized by an amino acid repeat domain with five to six amino acid repeats, in each of which a single HAMP domain was identified previously, but which are now known to comprise concatenated pairs of HAMP domains [25, 32, 33]. The function of these domains is not Thiamet G yet INCB28060 price clear, even though involvement in fungicide susceptibility and in osmosensing were suggested [19, 23, 25, 37]. Previous heterologous expression of truncated proteins, in which

several HAMP domains were deleted from group III HKs, i.e. from CaNik1p [25] and DhNik1p from D. hansenii[37], was not reported to result in inhibition of growth of the respective S. cerevisiae check details transformants. Whereas in the previous reports only selected HAMP domains were deleted, here we deleted all HAMP domains from CaNik1p (CaNik1pΔHAMP) and observed that the synthesis of this truncated protein in the transformed S. cerevisiae strain was associated with severe growth inhibition. This phenotype could be reversed by additional point mutation in the histidine phosphorylation site of the HisKA domain (H510) or by the expression of CaNIK1ΔHAMP in single gene deletion mutants of the response regulator SSK1 or of one of the components of the Hog1 module namely the MAP2K PBS2 and the MAPK HOG1. This proved that the inhibition of growth of the transformant upon expression of CaNIK1ΔHAMP was dependent on the functionality of both the histidine kinase activity of CaNik1p and the functionality of the Ssk1 – Pbs2 – Hog1 branch of the HOG pathway.

campestris gum operon Appl Environ Microbiol 1999, 65:278–282 Pu

campestris gum operon. Appl Environ Microbiol 1999, 65:278–282.PubMed 81. Kovach ME, Elzer PH, Hill DS, Robertson GT, Farris MA, Roop RM, Peterson KM: Four new derivatives of the broad-host-range cloning vector pBBR1MCS, carrying BI 2536 datasheet different

antibiotic-resistance cassettes. Gene 1995, 166:175–176.PubMedCrossRef Authors’ contributions MJ performed genetic analyses of the rosR mutants, carried out experiments concerning their phenotype characterization and plant experiments, and drafted the manuscript. JK conducted EPS and LPS analyses, TP performed microscope images and parameter analyses of biofilm. AS discussed the results and elaborated the final version of manuscript. All authors read and approved the final version of the manuscript.”
“Background see more Rhodocista centenaria, first described as Rhodospirillum centenum [1] is a thermotolerant phototrophic purple bacterium of the α-proteobacteria group isolated from

hot springs in Wyoming 1985. The slightly spiroid or vibrioid shaped cells are motile by means of a single long flagellum, their intracellular photosynthetic membranes are lamellar and their in vivo absorption spectra show features almost indistinguishable from those of Rhodospirillum rubrum [2]. However, 16S rRNA analysis elucidated considerable differences between the species, hence Rhodocista was separated into a new genus [3], now consisting of three

species [4, 5]. R. RAAS inhibitor centenaria is closely related to the plant-associated genus Azospirillum [6]. As virtually all phototrophic organisms, R. centenaria exhibits a sensory response to light originally described as “”Schreckbewegung”" [7]. Engelmann and also Manten [8] found that R. rubrum cells accumulated in the most intense area of light gradients between wavelengths 800 and 900 nm. R. centenaria shows a particularly unique form of macroscopic phototactic behaviour, first described in 1994 by Gest and coworkers [9]. On solid media, the phototactic colonies ASK1 move towards longwave light and away from light with wavelengths less than 650 nm [10]. R. centenaria develops lateral flagella in viscous media or on solidified surfaces. These flagella consist of a distinct flagellin whose expression is controlled by specific mot and fli genes [11]. For R. centenaria, a close relationship between chemotaxis and the phototactic response has been found [12]. As seen with many other photosynthetic bacteria, R. centenaria has multiple chemotaxis operons with distinct functions [13–15]. The chemotaxis gene cluster has been well characterized and most of the genes are similar to those of other Gram negative bacteria like Escherichia coli. In brief, the histidine kinase CheA is linked to the chemotactic receptors (MCPs) by the CheW protein [16].

025 g; Premabraze

616, Lucas-Milhaupt, Inc , Cudahy, CA,

025 g; Premabraze

616, Lucas-Milhaupt, Inc., Cudahy, CA, USA). The metal mixture binder is composed of 61.5 wt.% silver, 24 wt.% copper, and 14.5 wt.% indium micro- and nanoparticles. Metal wires such as copper, kovar, stainless steel (SUS), tungsten, silver, and titanium with a diameter of 1 mm were used as substrates of the emitters. One end of the metal wires was mechanically polished buy PD0332991 to have a flat surface. Around 0.5 μl of the CNT/metal binder mixture was put on a metal tip substrate. The CNT/metal binder mixture dried out very quickly in approximately 5 min due to high volatility of dichlorobenzene. Subsequently, an annealing process was carried out under LDN-193189 purchase vacuum at approximately 10−6 Torr at different temperatures. For comparison, a CNT emitter was prepared using silver nanoparticles (NPs; DGH, Advanced Nano Products Co., Ltd., Buyong-myeon, South Korea) under similar conditions. Figure 1 Schematics of the (a) CNT emitter fabrication process Ilomastat purchase and (b) experimental

setup for the characterization. The morphologies of the fabricated CNT emitters were characterized using a field emission scanning electron microscope (FESEM; Hitachi S-4800, Chiyoda-ku, Japan). The adhesive force of the CNT/metal binder coating on a substrate was measured by a pencil hardness test, which is described in American Society for Testing and Materials (ASTM) D3363. Field emission properties of the fabricated CNT emitters were characterized in a vacuum chamber, which is schematically shown in Figure  1b. A diode

type with a copper disc (diameter, 30 mm) acting as an anode was employed for the field emission test. A negative high voltage of 0 ~ −70 kV was applied to the CNT emitter while the Cu anode was grounded. The distance between the CNT emitter and the anode was fixed to 15 mm. In order to protect the high-voltage power supply due to high-voltage arcing, a current-limiting resistor (resistance, 10 MΩ) Vitamin B12 was installed between the power supply and the emitter. Results and discussion The role of metal binders is to attach CNTs to substrates. Silver NPs have been widely used for a metal binder due to good electrical conductivity and good contact with CNTs [3, 4, 28]. To investigate the performance as a binder, we prepared a CNT emitter on a tungsten metal tip (diameter, 1 mm) using silver NPs (Figure  2a). The annealing temperature to melt silver NPs was 750°C. As shown in Figure  2b, the fabricated CNT emitters exhibited very poor stability. Electron current density emitted from the emitter was initially 57.3 mA/cm2 at the applied voltage of 35.5 kV; however, the current density was dramatically reduced to 13.6 mA/cm2 for a 70-min operation (Figure  2b). Frequent arcing was observed during the test, and the emission current density was slowly decreased with the increase in the arcing events.

4535 (+1);

matched by mass alone); Note, modifications to

4535 (+1);

matched by mass alone); Note, modifications to sequence identified by MS. Peptides identified by MS are underlined in the protein sequence. Note the non-tryptic N-terminal peptide (958.5200 (+3) m/z), suggesting the methionine at position 7 is the true N-terminus. Cysteine residue potentially involved in disulfide bond/homodimer formation is marked with (*). Comparative gel-free proteomics of P. aeruginosa PAO1, PA14 and AES-1R using iTRAQ labelling and 2-DLC/MS-MS Proteins from stationary phase cultures of P. aeruginosa AES-1R, PAO1 and PA14 were proteolytically digested, labeled learn more using iTRAQ and analysed by 2-DLC-MS/MS. Multiple experiments were performed such that each strain was analysed in duplicate. Proteins with a ratio > 1.5 or < 0.67 (p-value < 0.05), and > 1.3 or < 0.77 (p-value < 0.01) were considered to be statistically differentially abundant. We identified a total of 1788 unique P. aeruginosa proteins, of which 1355 could be accurately quantified across the strains using iTRAQ. 162 proteins displayed significant differential abundance between the P. aeruginosa strains (Additional file 3). Of these, 60 were regulated identically between AES-1R compared to both PAO1 and PA14, 55 were only found in AES-1R versus PAO1, 39 were only found in AES-1R versus PA14 and 8 were differently abundant in AES-1R

compared to both PAO1 and PA14, but in the opposite direction (e.g. more Fedratinib molecular weight abundant in AES-1R compared to PAO1, but less abundant in AES-1R compared to PA14). Functional analysis of the differently abundant proteins showed they could be clustered into 6 major groups: i) virulence determinants (including proteins involved in iron acquisition, phenazine biosynthesis and secreted factors); ii) membrane-associated proteins (including proteins involved in transport, GPX6 antibiotic efflux, lipopolysaccharide (LPS) biosynthesis and outer membrane

proteins [OMP]); iii) transcriptional and regulatory proteins; iv) proteins involved in translation; v) find more metabolic proteins; and vi) proteins of no known function. Of the 123 proteins found to be significantly altered in abundance between AES-1R and PAO1, 83 were present at elevated abundance in the AES-1R strain (40 present at reduced abundance); while of the 105 proteins significantly altered in abundance between AES-1R and PA14, 73 were present at increased abundance in AES-1R (32 present at reduced abundance). Within the functional clusters, proteins could also be classified by their relative abundance when compared between strains. For example, proteins involved in translation (predominantly ribosomal proteins) were overwhelmingly more abundant in AES-1R than either PA14 or PAO1 (Additional file 3).