, 2012a, b). Pexidartinib manufacturer Although in some Mesorhizobium strains, no ACC deaminase activity was detected under free-living conditions (Ma et al., 2003b; Nascimento et al., 2012a), it has been shown that Mesorhizobium. sp. MAFF303099 expresses ACC deaminase under symbiotic conditions, in a NifA2-dependent manner (Uchiumi et al., 2004; Nukui et al., 2006). One explanation for these somewhat

disparate results includes the possibility that those acdS genes under the transcriptional control of a NifA-regulated promoter are either exclusively or primarily expressed within nodules resulting in a decreased rate of nodule senescence. On the other hand, those acdS genes under the transcriptional control of an Lrp-regulated promoter (Ma et al., 2003a)

are primarily involved in facilitating the nodulation process and are not expressed within the nodule itself. The aim of the present study was to assess the prevalence and phylogeny of acdS genes in www.selleckchem.com/products/MDV3100.html Mesorhizobium strains including isolates from a collection of chickpea mesorhizobia from Portuguese soils. In the present study, 12 Mesorhizobium type strains as well as 18 chickpea Mesorhizobium isolates from Portugal were tested for the presence of acdS genes and ACC deaminase activity under free-living conditions. The chickpea Mesorhizobium isolates from Portugal were collected from different sites throughout the country, as previously described (Alexandre et al., 2009). Mesorhizobium strains were grown at 28 °C in TY medium (Beringer, 1974), in YMA medium (Vincent, 1970), and in modified M9 minimal medium (Robertsen et al., 1981) when necessary. The bacterial strains used in this work are presented in Table 1. Mesorhizobium strains and isolates were grown in 5 mL of

TY medium at 28 °C for 2–4 days. The bacterial cultures were centrifuged at 16 000 g for 1 min and used for genomic DNA isolation using the E.Z.N.A bacterial DNA kit (Omega Bio-tek) following the manufacturer’s suggested protocol. To amplify the acdS gene next in Mesorhizobium type strains and chickpea Mesorhizobium isolates, PCR primers were designed based on the Mesorhizobium sp. MAFF303099 and M. ciceri bv. biserrulae WSM1271 acdS gene sequences, resulting in primers F2 (5′-CAAGCTGCGCAAGCTCGAATA-3′) and R6 (5′-CATCCCTTGC ATCGATTTGC-3′). The acdS gene was amplified in a ‘T Personal Cycler’ (Biometra) thermocycler using the following program: 3 min of initial denaturation at 95 °C, 35 cycles of 1 min denaturation at 94 °C, followed by 1 min and 30 s of primer annealing at 49 °C and 1 min of elongation at 72 °C, and a final elongation step of 5 min at 72 °C. The amplification product is a 760-bp fragment. After amplification, the PCR product was purified using the GFX DNA purification Kit (GE Healthcare, UK) and sequenced by Macrogen Inc. (Seoul, Korea). The obtained acdS gene sequences are presented in Table 1.

Fig. S1. Construction of M1-2 strain. Table S1. Nematodes studied. Please note: Wiley-Blackwell is not responsible for the content or functionality of any supporting materials supplied by the authors. Any queries (other than missing material) should be directed to the corresponding author for the article. “
“Freezing stool samples prior to DNA extraction and downstream analysis is widely used in metagenomic studies of the human microbiota but may affect the inferred community composition. In this study, DNA was extracted either directly

or following freeze storage of three homogenized human fecal samples using three different extraction methods. No consistent differences were observed in DNA yields between extractions on fresh and frozen samples; however, differences Selleckchem Epigenetic inhibitor were observed between extraction methods. Quantitative PCR analysis was subsequently performed on all DNA samples using six

different primer pairs targeting 16S rRNA genes of significant bacterial groups, and the community composition was evaluated by comparing specific ratios of the calculated abundances. In seven of nine cases, the Firmicutes to Bacteroidetes 16S rRNA gene ratio was significantly higher in fecal samples that had been frozen compared to identical samples that had not. This effect was further supported by qPCR analysis of bacterial groups within these two phyla. The results demonstrate that storage conditions of fecal samples may adversely affect the determined Firmicutes to Bacteroidetes ratio, which is a frequently used biomarker in gut microbiology. Investigating the composition of the human PD0325901 mouse microbiota and correlating findings to specific clinical or physiological states such as irritable bowel diseases and obesity has demonstrated the collective importance of the bacterial community present in the gut as a regulatory factor in health and disease (Young et al., 2011). Because of the large diversity and complexity of interactions between the resident bacteria,

various simplifications have been sought. An example of this is the use of the ratio between the two most Amino acid predominant phyla, namely the Firmicutes and the Bacteriodetes, as a biomarker in relation to human physiology (Armougom & Raoult, 2008; Guo et al., 2008; Mariat et al., 2009). Efficient and nonbiased extraction of total genomic bacterial DNA from the complex fecal samples is a crucial first step for many molecular-based studies of the bacterial community within the gut environment. Downstream applications, such as quantitative PCR and metagenomic sequencing, ultimately require unbiased DNA input to produce accurate and creditable research results. Previous studies have assessed the effectiveness of various DNA extraction procedures based on, for example, DNA yield, extent of DNA shearing, and use as template for subsequent PCR and have often been related to detection of specific pathogens (McOrist et al., 2002; Tang et al., 2008; Ariefdjohan et al.

Three-point amino acid substitutions, chosen on the basis of published data of HspH of B. japonicum (Lentze et al., 2003), were generated. Genetic manipulations involving O. oeni are unavailable, and so we produced

and studied all the proteins in E. coli. Among the three proteins analysed (Y107A, V113A and A123S), only A123S showed defective chaperone activity, as it prevented only around 60% of temperature-induced aggregation of the E. coli cellular proteins compared with native Lo18 WT. The results obtained for A123S www.selleckchem.com/products/E7080.html are in accordance with those reported for A109S by Lentze et al. (2003). By contrast, the results obtained for the two other proteins with amino acid substitutions were different from those obtained for HspH proteins. Y107A and V113A presented no significant modification in chaperone activity, in contrast to F94A/D and L100A, for which a lower activity was reported. Delmas et al. (2001) have shown that the native smHsp Lo18 is able to form dimeric, trimeric and oligomeric forms. These three multimeric structures were obtained after cross-linking experiments

either in vitro on purified Lo18 or in vivo Sotrastaurin using cells expressing Lo18 from O. oeni and E. coli. Our results showed no differences between the forms of the WT or Lo18 amino acid substitutions with monomeric, oligomeric and intermediate structures. Moreover, a relationship between the oligomerization process and chaperone activity has been suggested (Giese & Vierling, 2002; Gu et al., 2002). However, concerning the decreased chaperone activity Unoprostone of the A123S, no structural modification was demonstrated. Biochemical analysis of purified proteins may

provide information about differences in structural characteristics. Previous studies have shown that Lo18 WT is localized in the cytoplasmic and membrane fractions of heat-shocked cells of O. oeni (Jobin et al., 1997; Delmas et al., 2001). A similar distribution in both the cytoplasm and the membrane fractions was observed in E. coli expressing Lo18 WT and proteins with amino acid substitutions. The proportion of these heterologous proteins in the various fractions of the E. coli envelope was not explored. However, localization in the outer membrane fraction has been shown for the smHsp18 from Mycobacterium leprae expressed in E. coli (Lini et al., 2008). Our results obtained for membrane fluidity regulation in E. coli lead us to suggest that a major part of Lo18 is associated with the cytoplasmic membrane, even if we cannot exclude localization in other extracytoplasmic compartments. Among membrane-associated smHsp, those from the Mycobacterium genus (Cunningham & Spreadbury, 1998) are surface antigens, whereas Lo18, like smHsps from Synechocystis, shares a membrane-stabilizing activity in vitro (Török et al., 2001).

, 2006; Johnston et al., 2008). The mechanism of Neu5Ac transport in the Selleck Cobimetinib related organism Haemophilus ducreyi has also been characterized, and interestingly,

this utilizes an ATP-binding cassette (ABC) transporter (Post et al., 2005). Clearly, bacteria have evolved multiple mechanisms to capture this important molecule from their environment and it is likely that there are also additional families of bacterial transporters that have evolved to transport Neu5Ac. In this study, we use a ΔnanT strain of E. coli to enable a comparative study of two known Neu5Ac transporters and a third, previously uncharacterized, transporter from Salmonella enterica serovar Typhimurium (STm), which is a member of the sodium solute symporter (SSS) family, thus expanding the diversity of known Neu5Ac transporters in the prokaryotes. Lennox broth (LB) medium and M9 minimal medium (Neidhardt learn more et al., 1974) were used for routine and experimental growth of E. coli, respectively. General cloning was carried out in DH5α (Invitrogen).

All E. coli mutants constructed in this work for genetic studies are derivatives of the Keio collection wild-type strain BW25113 (Baba et al., 2006). The unmarked, in-frame ΔnanT mutant (referred to simply as ΔnanT) has been described in Mulligan et al. (2009) and was obtained by pCP20-mediated removal of the KanR cassette from the Keio collection strain JW3193, followed by plasmid curing (Datsenko & Wanner, 2000). Construction of strain SEVY1 (the ΔnanAT double mutant) has been described elsewhere (Mulligan et al., 2009). Neither of these strains grow on Neu5Ac as the sole carbon source,

Fludarabine molecular weight nor do they concentrate [14C]-Neu5Ac in an uptake assay, as reported for other nanT strains of E. coli (Vimr & Troy, 1985). Constructs pES1G and pES7 are derivatives of the low-copy-number vector pWKS30 (Wang & Kushner, 1991), and they carry, respectively, the E. coli MG1655 nanT gene and the H. influenzae Rd KW20 siaPQM operon under the control of an isopropyl-β-d-thiogalactopyranoside (IPTG)-inducible promoter; the construction of these plasmids has been described elsewhere (Mulligan et al., 2009). Plasmid pES41, carrying the STM1128 gene of STm strain LT2, was constructed in an equivalent manner using genomic DNA as a template in a PCR reaction using the oligonucleotides 5′-GCGGTACCGTAAAAGAAGGAGATATACATATGATTACACATTCTTTCGGC-3′ and 5′-GCGGATCCTTATAATGTCACCTTTGGTTCAGG-3′. Cells from starter cultures grown during the day in LB ampicillin (Amp) were harvested, washed twice in M9 minimal medium and diluted 100-fold in M9 Amp supplemented with 0.4% v/v glycerol for o/n growth at 37 °C. After three washes in M9, the o/n cultures were diluted to an OD650 nm of 0.1 in M9 Amp supplemented with Neu5Ac and IPTG, and their growth at 37 °C was followed hourly (IPTG was used at 1 mM and all sugars at 1 mg mL−1).

Isolated colonies were allowed to grow. C/EBP β expression was tested by western blotting in at least 10 different clones per plasmid transfection. Selected clones were grown in DMEM supplemented with 10% heat-inactivated fetal bovine serum, 2 mm glutamine, 100 units/mL penicillin, 0.1 mg/mL streptomycin, and 500 μg/mL G418 antibiotic (all chemicals were from Sigma-Aldrich). For western blot analysis of C/EBP β expression in CGNs, both samples from nucleo-cytosol separation

and total protein were used. To separately extract nucleic and cytosolic proteins from CGNs (Caruccio & Banerjee, 1999), CGN cell cultures from 12 rat pups were scraped, resuspended in 150 μL of extraction buffer with low salt [20 mm HEPES, pH 7.9, 10 mm NaCl, 3 mm MgCl2, 0.1% this website NP-40, 10% glycerol, 0.2 mm EDTA, 1 mm dithiothreitol (DTT), protease and phosphatase inhibitor cocktails] and left Birinapant on ice for 10–15 min with occasional tapping. Nuclei were pelleted by centrifugation at 700 g for 5 min at 4 °C. The cytoplasmic supernatant fraction

was transferred into another Eppendorf tube. Nuclei were washed with 200 μL of washing buffer to remove NP-40 (20 mm HEPES, pH 7.9, 0.2 mm EDTA, 20% glycerol, 1 mm DTT, protease and phosphatase inhibitor cocktails), and centrifuged at 700 g for 5 min at 4 °C. Nuclei were then resuspended in 60 μL of extraction buffer with salt (20 mm HEPES, pH 7.9, 400 mm NaCl, 0.2 mm EDTA, 20% glycerol, 1 mm DTT, protease and phosphatase inhibitor cocktails), and left on ice for 45 min with periodic mixing by tapping in order to extract nuclear proteins. Following centrifugation at 14 500 g for 15 min at 4 °C, supernatants were removed, aliquoted, quickly frozen on dry ice, and stored at −80 °C. The cytoplasmic fraction was further clarified by adding a one-third

volume of cytoplasmic extraction clarification buffer (20 mm HEPES, pH 7.9, 400 mm NaCl, 0.2 mm EDTA, 40% glycerol, 1 mm DTT, protease and phosphatase inhibitor cocktails) for 30 min at 4 °C in order to equilibrate the cytoplasmic proteins with NaCl, and this was followed by centrifugation at 14 500 g for 15 min. In parallel, total samples were obtained by directly harvesting cell cultures in Laemli Sample Buffer. 4��8C Transfected CGNs and DAOY stable clones were lysed in lysis buffer (50 mm Tris, pH 7.4, 1 mm EDTA, 1% sodium dodecyl sulfate, protease inhibitors), and then sonicated with a tip sonicator for 5 s at 10% power output. Total protein sample content was determined with the Lowry quantification method (Lowry et al., 1951), and 30 μg of each sample was loaded per lane for western blot analysis. Samples obtained as previously described were briefly sonicated and resolved by sodium dodecyl sulfate polyacrylamide gel electrophoresis before electroblotting. Membranes were incubated with antibodies against C/EBP β [rabbit (C-19) C-terminal rat epitope; Santa Cruz Biotechnology, Santa Cruz, CA, USA; cat. no.

Interestingly, members of the Burkholderia

cepacia complex that are inherently resistant to high concentrations of polymyxin B constitutively modify their lipopolysaccharide with l-Ara4N, and this modification is essential for cell viability (Loutet & Valvano, 2011). In contrast, Franscisella novicida uses a different strategy to resist polymyxin B; the lipid A phosphatase LpxF removes Stem Cell Compound Library cell assay the phosphate group at the 4′-position of lipid A (Wang et al., 2007). PmrC and CptA are phosphoethanolamine (pEtN) transferases that mediate the addition of pEtN to the 1 and 4′ phosphates of lipid A and to the phosphate of heptose 1 found in the lipopolysaccharide core, respectively (Gunn, 2008). Although these modifications have been shown to have a modest effect on S. Typhimurium resistance to polymyxin B, addition of pEtN to Neisseria gonorrhoeae and N. meningitidis lipid A greatly increased resistance to polymyxin B, LL-37, and protegrin (Tzeng et al., 2005; Lewis et al., 2009). Bacterial transporters are divided into importers and exporters belonging to different BIBW2992 ic50 families (Davidson et al., 2008). Members of the ABC transporter and the resistance-nodulation-division (RND) efflux pump families have been implicated

in AMP resistance. ABC importers, which usually rely on a periplasmic-binding protein, are believed to import AMPs from the periplasm or the periplasm–inner membrane interface into the cytosol, where they are most likely proteolytically Forskolin price degraded and recycled as nutrients (Fig. 1d). In contrast, exporters of the RND family are thought to export AMPs from the intracellular environment

into the extracellular environment. It appears most likely that RND pumps capture AMPs from the periplasm or from the periplasm–inner membrane interface, rather than from the cytoplasm. Export from the periplasm of various antibiotics that cannot cross the cytoplasmic membrane has been documented for RND pumps (Aires & Nikaido, 2005). The evidence for involvement of ABC transporters in AMP resistance came from the generation of strains in which the transporter genes were deleted or inactivated. These strains were more susceptible to AMPs than the wild-type strains, as judged by performing survival assays or determining minimum inhibitory concentrations. Screening for S. Typhimurium mutants hyper-susceptible to the AMP protamine led to the identification of the sapABCDF operon coding for the Sap (Sensitive to antimicrobial peptides) ABC importer (Parra-Lopez et al., 1993). In addition to protamine susceptibility, the sap mutants exhibited hypersensitivity to the bee-derived AMP melittin and crude extracts from human neutrophil granules.

In particular, slowly rising waveforms of light might activate the cells at different times because of differences in activation thresholds, making spike separation possible. To test this hypothesis, we compared the effects of sine wave patterns (5 Hz) versus short pulses of light (5 ms duration, every 1 s). The experiments were performed in the CA1 hippocampal region of rats using the optrode device shown in Fig. 2A. The effect of the two stimulation regimes could be seen on the wideband signal (Figs 4A and 5A). High-intensity light stimulation occasionally caused an artifactual potential via the photoelectric effect of the light on the conducting wires of the probe (Han et al.,

2009). This artifact CCR antagonist could also be detected in brain tissue without

ChR2 expression, such as the neocortex overlying the hippocampus, and could therefore be subtracted from the recorded signal. Following the implementation of spike detection and separation (Fig. 4C), the activation of several cells by the sine wave stimulus was readily detectable in the neurons’ spike raster plot (Fig. 4A), spike autocorrelograms (Fig. 4C; note the rhythmic oscillation at the 5 Hz stimulus frequency), and peristimulus spike time Epacadostat manufacturer histograms (Fig. 5C). Both the number of excited neurons and the magnitude of the responses increased with the intensity of the stimulus (Fig. 5C and D). In contrast, activation of clustered neurons by light pulses was often not detectable, even in neurons which showed a reliable response to the sine wave stimuli (Fig. 5C and D). This did not result from a failure of the light pulse to excite the neurons as waveforms of superimposed spikes were visible on the wideband signal during the pulses (Fig. 5B), and activation of

the network was obvious from the strong inhibitory responses of putative interneurons (Fig. 5C, fifth row). Instead, a failure to isolate the spikes triggered by the light pulses, due to superimposition of spike waveforms, is most probably the cause. Because the optical fiber terminated ∼ 100 μm above the recording Thiamine-diphosphate kinase sites (Fig. 2A), the stimulation was restricted to a small portion of the monitored tissue. As anticipated, the effect of the stimulation was typically observed on the shank carrying the optical fiber. This specificity was visible on both the wideband signals (Fig. 6A) and the responses of single neurons (Fig. 6B and D). At the low stimulus intensity of 50 μW, neuronal spikes were elicited only in neurons recorded by the shank with the optical fiber (Fig. 6B, left panel). After the intensity was raised to 100 μW, neurons recorded by the adjacent shank (250 μm away) could also be activated occasionally (Fig. 6B, right panel, and D). Either direct light activation or indirect synaptic activation could be the origin of these distant neurons responses, although occurrence of the latter should be rare given the sparsity of excitatory connections between CA1 pyramidal cells (Amaral & Witter, 1989).

A more complex analysis of the virological response to HIV treatment is used by the US Food and Drug Administration (FDA) for clinical trials

comparing the outcomes of two different treatment regimens [6]. There has, however, been little discussion in the literature about how best to measure virological response as a quality indicator, because the main use to date for this variable has been to compare the efficacies of different antiretroviral regimens. If an outcome selleck products indicator is to be useful for a measure of quality in clinical practice, it should fulfil a number of requirements in addition to correlating well with the patients’ future prognosis [4]. These characteristics include the ease and feasibility of collection and the degree to which the outcomes are predicted by differences in the provider characteristics rather than differences among individual patients. Our aim in this study was to describe the HIV virological response for a single health service using three different definitions of treatment failure and to discuss their relationship

to the requirements of a quality outcome measure. We included three measures of virological response, including the definition recommended by the US FDA, called SGI-1776 research buy the ‘time to loss of virologic response’ (TLOVR) algorithm [6]. The clinical data for this study were obtained for HIV-infected patients attending the Melbourne Sexual Health Centre between January 2000 and December 2008. During this period, 310 HIV-positive patients commenced antiretroviral isometheptene treatment for the first time (i.e. were antiretroviral naïve). The electronic medical record data, including laboratory measures and HIV treatment histories for each patient, were examined. Clinical files were reviewed to determine the reason for any change in HIV treatment. The outcomes of treatment were assessed using a number of different definitions of treatment failure. In the first analysis (definition 1), we used

the TLOVR algorithm, where an individual is deemed to have failed if a plasma HIV-1 RNA level <400 copies/mL was never achieved, or they had confirmed virological rebound from <400 copies/mL on two consecutive readings, or they had discontinued their first treatment regimen for any reason [6]. In the second analysis (definition 2), an individual was deemed to have failed if the plasma HIV-1 RNA was never below 400 copies/mL, or their viral load rebounded above 400 copies/mL (on two consecutive readings) while on any treatment. They were permitted to change treatment so long as their viral load remained below 400 copies/mL and were also permitted to stop treatment as long as their last viral load on treatment was below 400 copies/mL.

Ca2+ increased the efficacy of

tetronasin, as would be predicted, but Na+ was almost as effective, despite the affinity of tetronasin for Na+ being < 5% that for Ca2+ Etoposide (Grandjean & Laszlo, 1983). In general, however, the effects of changing cation concentrations were relatively small and some could not be explained simply by the reported ion specificity of the ionophores. One possible cause of the small response was most likely the relatively small changes in concentration and therefore ionic gradient that were considered feasible, based on what might be achieved in vivo. The increase in [Na+] was only 26%, which would have a small effect on the transmembrane Na+ gradient. However, the change in [Ca2+] was substantial, a 2.6-fold increase, yet potentiation of tetronasin was still small. Several studies have been made previously, with some success, to apply the principle of cation enhancement of ionophores with ruminal bacteria and ruminal fermentation. Rumpler et al. (1986) found that adding Na+ to the diet of steers receiving monensin or lasalocid caused methane production to be decreased. This result was therefore consistent with the main mode of action of monensin as it is presently understood (Russell, 1987), but not with the K+/H+ exchange mechanism proposed for lasalocid (Schwingel et al., 1989). Increasing [K+] increased the potency of monensin towards ruminal bacteria in vitro

(Dawson & Boling, 1987), which

might not be expected to occur if the direction of induced K+ flux was outward, as in the Russell GSK2126458 cell line (1987) scheme. Chirase et al. (1987) observed a significant interaction between K+ and lasalocid in continuous cultures, but also Mg2+ and monensin or lasalocid despite the low affinity of these ionophores for divalent ions. Thus, although interactions undoubtedly occur between the concentrations of individual cations and the efficacy of ionophores, their magnitude and direction do not always appear to correspond to known ionophore specificity see more and the magnitude and direction of transmembrane ion gradients that have been measured in ruminal bacteria. Furthermore, the effects of combinations of cations and ionophores appeared to be species dependent, possibly indicating that transmembrane ion gradients are different in different rumen bacterial species. The measurements of protonmotive force and ATP pools in E. ruminantium may help to explain some of these observations. Despite a rapid inhibition of cell growth, only relatively minor changes in intracellular cation concentrations were seen when monensin or tetronasin was added to the culture. Some efflux of Na+ and K+ was induced by monensin and Ca2+ by tetronasin. Undoubtedly, the measured ion concentrations in whole cells may not reflect the concentration of ions free in solution; cell walls, proteins and nucleic acids would be expected to bind Na+, K+ and Ca2+.

Previous studies on S. aureus demonstrated differential expression of a variety of genes in biofilm as compared to planktonic phase. Genes-encoding proteins associated with cell attachment, fibrinogen-binding proteins, staphylococcal accessory regulator A protein (SarA) and proteins involved in PIA synthesis are up-regulated. In contrast, proteins such as proteases and toxins are down-regulated (Resch et al., 2005, 2006). We studied PIA content in planktonic Enzalutamide in vitro and biofilm preparations that we used,

as PIA is the main structural component of the biofilm state in many bacteria. Our data show that planktonic phase bacteria have minimal PIA quantities. Although biofilm state bacteria exhibit a number of phenotypic characteristics, PIA synthesis seems to contribute to some extent to resistance of biofilm phase bacteria buy CYC202 to immune system responses and may contribute to the chronic and silent course of biofilm-associated infections (Cerca et al., 2006). A prerequisite for infection elimination is interaction between the host cells and the pathogen or pathogen-derived material. Here, we demonstrated that macrophages efficiently phagocytose

biofilm bacteria, but nevertheless, eradication of infection cannot be achieved. Inefficient killing of phagocytosed bacteria along with impaired Th1 immune response reflects this finding. Biofilm bacteria persist intracellularly and modulate immune responses to their favour. We thank the Advanced Light Microscopy facility of the Medical School, University of Patras, for their support with immunofluorescence and phase contrast experiments. “
“Phosphate signaling and acquisition are critical for the bacterial response to phosphate limitation, and bacteria express multiple factors to scavenge phosphate. We previously found that multidrug-resistant strains of Pseudomonas aeruginosa from critically ill patients can form unusual outer-surface appendages harboring PstS proteins. Here, we have expanded our investigation to DING proteins that like PstS belong to the family of high-affinity phosphate-binding

proteins but have strong similarity with eukaryotic DING proteins. We demonstrate the localization of DING on PstS-containing outer-surface appendages in both multidrug-resistant strain Calpain MDR25 and the PA14 strain of P. aeruginosa. However, the number of cells producing appendages and the amount of appendages on each cell in PA14 were found to be negligible, unless overexpression of either PstS or DING was achieved by transformation with constructed plasmids. We further noticed that DING expression under low phosphate conditions was significantly higher in MDR25 compared to PA14 which may explain the greater abundance of appendages in MDR25. Our finding that DING proteins are localized on extracellular appendages provides an opportunity to study the interaction of bacterial DING with host proteins by mimicking the action of host DINGs. “
“Streptococcus suis serotype 2 (SS2) is an emerging zoonotic pathogen.