Acta Stomatol Belg 1992,89(3):155–162.PubMed 20. Germaine GR, Tellefson LM: Effect of human saliva on glucose uptake by Streptococcus mutans and other oral microorganisms. Infect Immun 1981,31(2):598–607.PubMed 21. Mansson-Rahemtulla B, Baldone DC, Pruitt KM, Rahemtulla F: Effects of variations in pH and hypothiocyanite concentrations

on S. mutans glucose metabolism. J Dent Res 1987,66(2):486–491.CrossRefPubMed 22. Tenovuo J, Anttilla O, Lumikari M, Sievers G: Antibacterial effect of myeloperoxidase against Streptococcus mutans. Oral Microbiol Immunol 1988,3(2):68–71.CrossRefPubMed 23. NU7026 mouse Lumikari M, Soukka T, Nurmio S, Tenovuo J: Inhibition of the growth of Streptococcus mutans, Streptococcus sobrinus and Lactobacillus casei by oral peroxidase systems in human saliva. Arch Oral Biol 1991,36(2):155–160.CrossRefPubMed 24. Lenander-Lumikari M: Inhibition of Candida albicans bythe Peroxidase/SCN-/H2O2 system. Oral Microbiol Immunol 1992,7(5):315–320.CrossRefPubMed 25. Mikola H, Waris M, Tenovuo J: Inhibition of herpes simplex virus type 1, respiratory syncytial virus and echovirus type 11 by peroxidase-generated hypothiocyanite. Antiviral Res 1995,26(2):161–171.CrossRefPubMed 26. Tenovuo J, Makinen KK: Concentration of thiocyanate and ionizable iodine in saliva of smokers and nonsmokers. J Dent Res 1976,55(4):661–663.CrossRefPubMed 27. Lamberts BL, Pruitt

KM, Pederson ED, Golding MP: Comparison of salivary peroxidase system components in caries-free and caries-active naval recruits. Caries Res 1984,18(6):488–494.CrossRefPubMed 28. Pruitt KM, Tenovuo J, Fleming W, Adamson M: Limiting factors for the generation of hypothiocyanite ion, an check details antimicrobial agent, in human saliva. Caries Res 1982,16(4):315–323.CrossRefPubMed 29. Caspase inhibitor Thomas EL, Milligan TW, Joyner RE, Jefferson MM: Antibacterial activity of hydrogen Verteporfin peroxide and the lactoperoxidase-hydrogen peroxide-thiocyanate system against oral streptococci. Infect Immun 1994,62(2):529–535.PubMed 30. Thomas EL, Jefferson MM, Joyner RE, Cook GS, King CC: Leukocyte myeloperoxidase and salivary lactoperoxidase: identification and quantitation in human mixed saliva. J Dent Res 1994,73(2):544–555.PubMed 31. Adolphe Y, Jacquot M, Linder M, Revol-Junelles

AM, Milliere JB: Optimization of the components concentrations of the lactoperoxidase system by RSM. J Appl Microbiol 2006,100(5):1034–1042.CrossRefPubMed 32. Rosin M, Kocher T, Kramer A: Effects of SCN-/H2O2 combinations in dentifrices on plaque and gingivitis. J Clin Periodontol 2001,28(3):270–276.CrossRefPubMed 33. Rosin M, Kramer A, Bradtke D, Richter G, Kocher T: The effect of a SCN-/H>2O2 toothpaste compared to a commercially available triclosan-containing toothpaste on oral hygiene and gingival health – a 6-month home-use study. J Clin Periodontol 2002,29(12):1086–1091.CrossRefPubMed 34. EN 1040 Chemical disinfectants and antiseptics. Basic bactericidal activity. Test method and requirements (phase 1)Beuth-Publishing, Berlin 1997. 35.

The predominant spoligotype, widely dispersed geographically (Table 1 &2), was found in the international data base to have a pattern with a spoligotype number SB0120 with the corresponding hexacode of 6F-5F-5F-7F-FF-60. Five out of the six study districts had this predominant spoligotype, and Namwala district accounted for 30% of spoligotype SB0120. The second most predominant spoligotype had a pattern named SB0871 with a corresponding hexacode of 6F-4F-5F-7F-FF-60. Isolates C14 was

named SB1572 with a hexacode number of 6F-5F-5F-7F-FF-40, isolate C16 was SB1536 with a hexacode number of 2F-5F-5F-6F-FF-60 and isolate C19 was SB0162 with a hexacode number of 00-00-00-0F-FF-60. The distribution

of these spoligotypes on the international data base is shown in Table 2. Table 2 Major Spoligotypes in Zambia selleck chemicals Spoligotype1 Shared type2 Geographical distribution Sp1 SB0120 France, Belgium, Brazil, South Africa, Sri Lanka, Iran, The Netherlands, Spain, China, Japan, Portugal, Russia, Denmark, Zambia Sp2 SB0871 France Sp3 SB1763* Zambia Sp4 SB1764* Zambia Sp5 SB1572 Italy Sp6 Foretinib SB1765* Zambia Sp7 SB1536 Italy Sp8 SB1766* Zambia Sp9 SB1767* Zambia Sp10 SB0162 Belgium 1 Arbitrary spoligotype designation 2 Shared type, designation of the spoligotype in the World Spoligotype Database. *New Spoligotype assigned by http://​www.​mbovis.​org Five individually www.selleckchem.com/products/salubrinal.html occurring isolates (16.1%) displayed new spoligo patterns that have not yet been described on the international second spoligotyping data base (Figure 2 and Table 2). These isolates

originated from Namwala district (isolate C26, 42 and C41); from Mumbwa (isolate C21); and from Monze (isolate C9) (Table 1 and Figure 2). These new patterns were allotted new spoligo numbers as SB1763 (hex code 66-03-5F-6D-FF-60), SB1764 (hex code 60-0F-1F-6C-FF-00), SB1765 (hex code 2F-5F-5F-7F-FF-40), SB1766 (hex code 6F-4F-1F-6F-FF-60) and SB1767 (hex code 62-0E-50-09-FF-40) by http://​www.​mbovis.​org Table 2. The technique showed a good discrimination power; Hunter Gaston Discriminatory Index (HGDI = 0.98) (Table 1 and Figure 2.). Discussion Our results do not agree with what has been found in other parts of Africa [21, 22], where more than 40% of the animals with tuberculous lesions had Non-tuberculous Mycobacteria (NTM). In this study, only two animals had mycobacteria other than M. bovis. However, our findings tie up with a similar study conducted in Algeria [23]. Whereas excluding the differences in bacterial species as accounting for these observations [23], strain isolation has been found to be dependant on the specific type of media used [24]. The usage of specific culture media such as Stonebrink has been shown to increase the recovery and discrimination of strains on culture [25, 26].

5 eV), which can be ascribed to the trap states near the film surface.

The S parameter of the injection energy was approximately between 0.5 and 2 keV, which mainly represented the annihilation events occurring in the aluminum oxide film. Figure 5a shows that the S parameter initially increased rapidly, which indicated a higher vacancy defect density of the inner oxide film than that of the surface. A decrease was observed beyond 1 keV, demonstrating that the S parameter of the Al2O3/Si interface was lower than that of the Al2O3 films. The lower S parameter can be attributed to the positron annihilation with high-momentum electrons of oxygen at the interface. This result was probably due to the SiO x layer grown between the aluminum oxide and Si substrate, which reportedly has an important click here function in excellent surface passivation [6, 20, 21]. https://www.selleckchem.com/products/sch772984.html The S parameter continued to increase after 2 keV with increased incident energy because larger

portions of positrons were injected into the silicon substrate. The S parameter in the substrate was much higher than that in the oxide film because of the different chemical environments of annihilation. The S parameter did not reach a constant value before 10 keV, which implied that positrons with 10 keV energy check details cannot completely penetrate the Si substrate far from the oxide layer. The S-E plot in Figure 5a also shows that the S parameter in Al2O3 films (about 1 keV) evidently decreased with increased annealing temperature because of the decreased density of trap vacancies in the Al2O3 films. The W parameter was more sensitive to the chemical environment of the annihilation site. The larger W and smaller S parameters indicated more positrons

annihilating core electrons. Thus, the smallest S and largest W parameters of the sample annealed at 750°C (Figure 5a,b) implied that the Al2O3 films had been compressed at this temperature with the lowest vacancy defect density and that the film structure probably did not change. Figure 5 Doppler broadening spectroscopy of S – W parameters vs. positron incident energy. (a) S and (b) W parameters vs. positron incident energy for samples annealed at different temperatures for 10 min. (c) S-W plot for samples annealed at different temperatures for 10 min. The S and W parameters of the same incident energy were plotted in one graph, as shown in Figure 5c. The Dimethyl sulfoxide S vs. W diagrams of monolithic materials present clusters of points because all S or W parameters are almost the same [14]. For example, in one type of defect, the S and W parameters may vary with the positron incident energy, and the S-W plot extends to the line passing the data point of the bulk region without defect [13, 14]. The slope of the line changes with the layers of different compositions and defect types. Thus, the annealed sample consisted of a three-layered structure in which each curve consisted of three extended line segments (Figure 5c).

CS settled the mesocosm experiment and assisted in the samplings. EGB, MB, FP and AM conceived the idea and contributed in performing part of the analyses and in drafting the manuscript. All authors have given final approval Bafilomycin A1 supplier of the version to be published.”
“Background Yersinia pestis and Bacillus anthracis are two pathogens of significant concern to public health from a biodefense perspective [1, 2]. Y. pestis, the causative agent of plague, is a Gram-negative, highly communicable coccobacillus that has been responsible for three historic pandemics with high mortality rates [3–5]. The microorganism possesses a Type III secretion mechanism common to several

human, animal and plant pathogens, whereby a series of pathogen-specific structural proteins form a syringe-like structure capable of injecting virulence factors into the mammalian host cell.

These virulence factors then facilitate pathogen use of host nutrients and thwart the host immune response, ultimately causing cell and host death [6, 7]. Naturally occurring plague can be transmitted from infected fleas and rodents to humans, and although the pathogen can be phagocytosed, it can also resist destruction by manipulating the host defense mechanism(s), potentially GSK872 through antigenic mimicry [8]. Y. pestis then multiplies rapidly leading to necrosis of lymph nodes, a condition known LY2874455 ic50 as bubonic plague, which can result in death if untreated [2]. In some cases the infection can spread through the blood stream resulting in systemic plague (septicemia) or to the lungs resulting next in the highly contagious and deadly form of the disease known as pneumonic plague. There are currently no rapid, widely available diagnostic tests for plague, and the most common treatment is streptomycin [2,

3], an antibiotic with adverse effects. Two other species from the genus Yersinia are also human pathogens: Y. pseudotuberculosis and Y. enterocolitica[9, 10]. Despite their high degree of sequence similarity to Y. pestis, these two near neighbors of Y. pestis manifest in very different symptoms, ranging from abdominal pain to septicemia in humans, usually caused by infection through contaminated food. Infections caused by Y. pseudotuberculosis or Y. enterocolitica can be effectively treated with antibiotics and in most cases are self-limiting. Notably, Y. pestis is reported to have evolved from Y. pseudotuberculosis within the past 10,000 years [11]. B. anthracis is a Gram-positive, rod-shaped spore-forming bacterial pathogen and the causative agent of anthrax [12, 13]. Human, livestock, and wildlife mortalities attributable to anthrax occur in numerous regions of the world, although the majority of cases are found in less industrialized nations [14]. Three forms of the disease have been described: cutaneous, intestinal and inhalational.

In addition, the diameter of the Ge/GeO x nanofilaments (or NWs) of approximately 40 nm is calculated using a new method under SET. The low-current operation of this RRAM device will make it useful in nanoscale nonvolatile memory applications. Methods Ge NWs

were grown by the VLS Epacadostat supplier technique using Ge powder as the starting material (purity of 99.999%). Silicon (Si) wafers with an ultrathin gold (Au) coating as a catalyst were used as substrates. The substrate was annealed at 600°C for 30 min in a vacuum chamber to form isolated Au nanoparticles (NPs), or commercial Au NPs were used as substrates to grow NWs. The typical diameter of the Au NPs was approximately 5 nm, which was determined by scanning electron microscopy (SEM) (Figure 1a). Ge powder was placed in an alumina selleck screening library boat and inserted in a horizontal tube furnace. The furnace was heated at 900°C for 30 min under argon with a flow rate of 10

sccm to grow NWs through the VLS technique. High-density Ge NWs with a diameter of approximately 100 nm and length of approximately 100 μm were observed by SEM (Figure 1b). The Ge NWs possessed a core-shell structure, ABT-737 order as shown in the transmission electron microscopy (TEM) image in Figure 1c. This suggests that the core region is Ge-rich, and the shell region is oxygen-rich, i.e., GeO x . It is expected that the GeO x layer will contain more defects than the Ge-rich core, which may be useful for resistive switching memory applications. The defects in the Ge/GeO x NWs were observed by both XPS and PL (Figures 2 and 3). PL measurements were obtained on a Triax 320 monochromator (Jobin Yvon, Edison, NJ, USA) and photomultiplier detector with an excitation wavelength of 325 nm. Figure 1 SEM and TEM images. SEM images of (a) Au nanoparticles and (b) Ge NWs on Si substrates. (c) TEM image of core-shell Ge/GeO x NWs. Figure 2 XPS spectra of Ge 3 d core-level

electrons of the Ge/GeO x NWs. Figure 3 PL and deconvoluted spectra. PL spectra of the Ge/GeO x NWs (a) measured at temperatures of 10 to 300 K and (b) deconvoluted spectra at 300 K. Defects in the Ge/GeO x NWs and resistive switching memory characteristics were also assessed by fabricating an IrO x /Al2O3/Ge NWs/SiO2/Si Olopatadine MOS structure, as shown in Figure 4a. MOS capacitors were fabricated using a shadow mask to pattern IrO x electrodes onto Al2O3 that was grown on dispersed Ge/GeO x NWs. The memory device consisted of three stacked layers: a top tunneling layer of Al2O3 (10 nm), a defect-rich Ge NW layer, and a thin tunneling layer of SiO2 (approximately 4 nm). After cleaning the Si wafer, an SiO2 layer was grown by annealing in a hot furnace as described above. The Ge/GeO x NWs were then dispersed on the SiO2/Si substrate. To deposit the TE of IrO x , a thin layer of Al2O3 was also deposited.

Four measurements were taken in each independent analysis, with each measurement consisting of six runs, each lasting 10 s. The average from each of these measurements was calculated using Zetasizer series software 6.20 (Malvern Instrument). The instrument was set to automatically Histone Acetyltransferase inhibitor select the best conditions for measurements. A kinetic study was not performed in EMEM/S+ because of evidence of a stable suspension from time 0 to 24 h under exposure conditions when serum was present. Zeta potential Zeta potential measurements were performed to determine the stability of the PBH-capped AuNPs in Milli-Q water and URMC-099 research buy in the different medium suspensions

(EMEM/S+ and EMEM/S-). A Malvern Zetasizer Nano-ZS and folded capillary cells (Malvern Instruments Ltd., Worcestershire, UK) were used. One-millilitre aliquots of AuNP suspensions (100 μg/ml) were taken directly after preparation and 24 h after incubation in the different media. Due to the limitations of high salt content in both medium suspensions, zeta potential measurements were performed only in Milli-Q water. Three independent measurements were taken, and the mean ± SD is presented. Optical selleck products microscopy and visual sedimentation of AuNP suspensions An inverted light microscope Axiovert 25 (Carl Zeiss, Madrid, Spain) equipped with a Canon EOS 1000D (Canon, Madrid, Spain) camera was used to take images. NP suspensions (0.781 to 100

μg/ml) were prepared in EMEM/S+ and EMEM/S- medium, and 100-μl aliquots of each concentration were suspended in 96-well plates. Suspensions were viewed 24 h after incubation in exposure conditions (37°C/5% CO2). A recent study carried out by Cho et al. [40] highlights the importance Terminal deoxynucleotidyl transferase of considering sedimentation when carrying out NP toxicity studies in vitro. Those authors reported that different concentrations of NPs in the bottom of culture plates or ‘interaction zones’ caused by distinct ratios of sedimentation to diffusion velocities can result in variations in uptake. To detect differences in dispersion and sedimentation

between the PBH-capped AuNPs in EMEM/S+ and EMEMS/S- medium, photographs were taken of the AuNP suspensions (100 μg/ml) in 1.5-ml tubes after 24-h incubation under exposure conditions. Cell culture and AuNP exposure Human liver hepatocellular carcinoma cells (Hep G2) were from the American Type Culture Collection (Manassas, VA, USA). These cells were cultured in EMEM medium supplemented with 10% FBS, 1% penicillin/streptomycin, 1% ultraglutamine and 1% NEAA. They were incubated at 37°C with 5% CO2 in a humidified incubator. For AuNP exposure, cells were plated at densities of 7.5 × 104 cells per millilitre in 96-well tissue culture microtiter plates (Greiner-Bio one, CellStar, Madrid, Spain) and subsequently incubated for 24 h. After this period, cells were exposed to a series of concentrations of the five AuNP preparations for either 2 or 24 h for ROS production studies or for 24 or 48 h for the cytotoxicity studies.

Nucl Acids Res 2005, 33:244–248.CrossRef 55. Sali A, Potterton L, Yuan F, Van Vlijmen H, Karplus M: Evaluation of comparative

protein modeling by MODELLER. Proteins 1995,23(3):318–326.PubMedCrossRef Authors’ contributions All authors reviewed and approved the final version Emricasan purchase of the manuscript. LR and PG conducted the protein analysis. YZ performed bioinformatics analyses. DMD supervised the work in USA. PG, BM and AR designed the study, obtained funding and wrote the manuscript. Competing interests The authors declare that they have no competing interests.”
“Background Francisella tularensis is a highly clonal, recently-emerged pathogen that causes tularemia, which presents in several main forms: pneumonic (30%-60% mortality), ulceroglandular, and oropharyngeal [1]. The latter two are associated with lower mortality. F. tularensis is currently divided into three subspecies (tularensis, holarctica and mediasiatica), with F. novicida recognized as a very closely related species, or as

another subspecies by some authors [2–4]. These taxa vary in virulence, geographic distribution, overall genetic diversity, and host/vector associations [3, 5–9]. Human tularemia is a disease at which the clinical severity depends upon the route of infection, subspecies of the infection strain, and timely therapeutic response [9]. Cases in Europe are caused by F. tularensis subsp. Selleck Brigatinib holarctica, and in many rural areas of the Balkans and countries further east outbreaks are water-borne, resulting in oropharyngeal tularemia [10–12]. No known cases by F. tularensis subsp. mediasiatica are known and only a few by F. novicida have been documented [13, 14]. F. tularensis subsp. tularensis is restricted to North

Rebamipide America, whereas F. tularensis subsp. holarctica is found throughout the Northern Hemisphere [3, 15]. Despite its wider geographic distribution F. tularensis subsp. holarctica has markedly lower genetic diversity than F. tularensis subsp. tularensis [5, 7, 8]. Significant gains toward deciphering the evolutionary history of F. tularensis overall and, in particular, F. tularensis subsp. holarctica have been made by using whole genome comparisons for single nucleotide polymorphism (SNP) discovery coupled with subsequent canonical SNP (canSNP) analysis [15, 16]. Numerous new groups were identified within F. tularensis subsp. holarctica (Figure 1A) [15, 16], two of which, B.Br.013 (includes subclades B.Br.013/014 and B.Br.LVS in [15]) and B.Br.FTNF002-00, were predominant in Europe but geographically segregated [15]. In the Western European countries of Spain, France, and Switzerland selleckchem almost all isolates belong to the highly monomorphic B.Br.FTNF002-00 group [15–18]. In contrast, in large portions of Central and Eastern Europe, from the Czech Republic to Russia, most F. tularensis subsp. holarctica isolates are assigned to various lineages within the B.Br.013 group [15, 16]. Figure 1 Phylogenies of Francisella tularensis subsp. holarctica.

The new eae sequences of strains analyzed were deposited in the European Bioinformatics Institute (EMBL Nucleotide Sequence Database). Quantitative invasion assay Quantitative assessment of bacterial invasion was performed as described previously [53] with modifications. Briefly, washed HeLa and polarized and differentiated T84 cells were infected with 107 colony-forming click here units (c.f.u.) of each aEPEC strain for 6 h or 3 h for tEPEC E2348/69. The different incubation-periods used were due to the more

efficient colonization of tEPEC in comparison with the aEPEC strains; moreover, tEPEC E2348/69 induced cell-detachment in 6 h. Thereafter, cell monolayers were washed five times with PBS, and lysed in 1% Triton X-100 for 30 min at 37°C. Following cell lysis, bacteria were re-suspended in PBS and quantified by plating serial dilutions onto MacConkey agar plates to obtain the total number of cell-associated bacteria (TB). To obtain the number of intracellular bacteria (IB), a Selleckchem ACP-196 second set of infected wells was washed five times and 4SC-202 further incubated in fresh media with 100 μg/mL of gentamicin for one hour. Following this incubation period, cells were washed five times, lysed with 1% Triton X-100 and re-suspended in PBS for quantification by plating serial dilutions. The invasion indexes were calculated as the percentage of the total number of cell-associated bacteria (TB) that

was located in the intracellular compartment (IB) after 6 h (or 3 h for tEPEC E2348/69) (IBx100/TB) of infection. Assays were carried out in duplicate, and the results from at least three independent experiments were expressed as the percentage of invasion Cyclic nucleotide phosphodiesterase (mean ± standard error). Cytoskeleton polymerization inhibitor In order

to evaluate the participation of cytoskeleton components in the invasion of aEPEC 1551-2, HeLa cell monolayers were incubated with 1 and 5 μg/mL of Cytochalasin-D or Colchicine (Sigma-Aldrich, St. Louis, MO) 60 min prior to bacterial inoculation [33]. After that, cells were washed three times with PBS and the invasion assay was performed as described above. S. enterica sv Typhimurium and S. flexneri were used as controls. EGTA treatment for tight junction disruption In order to evaluate the interaction of aEPEC 1551-2 with the basolateral surfaces of T84 cells, differentiated cell monolayers (14 days) were incubated with 1 or 5 mM of EGTA (Sigma-Aldrich, St. Louis, MO) 60 min prior to bacterial inoculation [35]. After that, cells were washed three times with PBS and the invasion assay was performed as describe above. S. enterica sv Typhimurium and S. flexneri were used as controls. Detection of actin aggregation To detect actin aggregation the Fluorescence Actin Staining (FAS) assay was performed as described previously [12]. Briefly, cell monolayers were infected for 3 h, washed three times with PBS and incubated for further 3 h with fresh medium.

2000). This

is followed by the formation of diacetylene, triacetylene, and benzene (Cernicharo et al. 2001). In the following evolutionary stage (called the proto-planetary nebulae phase), the first spectral signatures of ICG-001 nmr aromatic compounds appear. The 3.3, 6.2, 7.7, 8.6, and 11.3 μm aromatic stretching and bending modes first make their appearance during the proto-planetary nebulae phase, and are found to become stronger in the subsequent planetary nebulae (Kwok 2000) phase (Fig. 1). These aromatic features are accompanied by aliphatic features at 3.4 and 6.9 μm selleck chemicals in the spectra of proto-planetary nebulae. The detection of out-of-plane C-H bending modes at 12.1, 12.4, and 13.3 μm suggests that the aromatic rings are not all connected to each other and there are many exposed edges of the rings (Kwok et al. 1999). Also present in the spectra of proto-planetary nebulae are broad plateau emission features at 8 and 12 μm which are due to collections of in-plane and out-of-plane bending modes of aliphatic chains (Kwok et al. 2001). Fig. 1 Infrared Space Observatory spectrum of the planetary nebula NGC 7027 superimposed on the Hubble Space Telescope image of the object. The aromatic infrared

bands (AIB) are marked in green. The identifications of these bands are given in the legend. The lines labeled in purple are atomic lines. The strengths of the AIBs show that aromatic below compounds are being produced in large quantities These observations suggest that even under the extremely selleckchem low density environment of the circumstellar envelopes, complex organics can be synthesized. One possible scenario is that

starting from acetylene, these linear molecules bend to form benzene, and all kinds of aliphatic chains get attached to the rings. The aromatic rings grow in size, possibly as the result of photochemistry. Since we know the evolutionary and dynamical timescales of the AGB (~104 yr), proto-planetary nebulae (~103 yr), and planetary nebulae (~104 yr) stages, these time scales constrains the chemical timescales that the synthesis must take place. Circumstellar molecular synthesis is therefore extremely efficient (Kwok 2004). It is interesting to note that these spectral characteristics resemble the infrared spectra seen in coal (Guillois et al. 1996), kerogen (Papoular 2001), soot (Pino et al. 2008), and petroleum (Cataldo et al. 2004). What all these compounds have in common is that they are all disorganized organic matter with mixed sp2/sp3 structures. While coal, kerogen and petroleum are remnants of life on Earth, the carbonaceous grains produced by stars condensed directly from the gas phase (similar to the formation process of soot) and are probably amorphous nanoparticles with a few aromatic islands connected by aliphatic chains (Kwok and Zhang 2011).

Colonization of rice plants was evaluated in vivo using a rifampicin-resistant mutant of strain REICA_142T, denoted REICA_142TR. The mutant was selected on R2A agar medium amended with 25 μg ml-1 rifampicin (Sigma-Aldrich, St. Louis, MO) and streaked to purity. One-day-old germinated rice seeds

were incubated for 1 h with 8.4 log cells of REICA_142TR CFU ml-1 (REICA_142TR treatment) or with sterile phosphate buffer solution (pH 6.5; control treatment) [44]. For each treatment, four replicate rice seedlings were grown in autoclaved as Protein Tyrosine Kinase inhibitor well as natural V soil [45] for up to 4 weeks at 70% water holding capacity. Water lost from the pots was replaced daily using sterile demineralized water. Following growth, all rice plants were surface-sterilized [46], rice tissue was treated with mortar and pestle, after which serial dilutions of the resulting click here homogenates were made and plated onto selective agar (R2A supplemented with Rif). Following plate incubations at 28°C for 72 h, the bacterial communities obtained from the plant tissue were enumerated. The ability of strain REICA_142TR to invade rice plants from the V soil was thus confirmed by isolating colonies from the relevant plates (at least one per replicate) and performing BOX-A1R

PCR on these [47]. Availability of supporting data The accession numbers for the 16S rRNA gene find more sequences of Enterobacter oryziphilus strains REICA_084, REICA_142T and REICA_191 are [GenBank:JF795012, JF795013, JF795014], and of Enterobacter oryzendophyticus strains REICA_032, REICA_082T and REICA_211 are [GenBank:JF795010, JF795011, JF795015], ID-8 respectively. The accession numbers for the rpoB gene sequences of strains REICA_084, REICA_142T

and REICA_191 are JF795018, JF795019 and JF795020, and of Enterobacter oryzendophyticus strains REICA_032, REICA_082T and REICA_211 are JF795016, JF795017 and JF795021, respectively. The generated phylogenetic trees from the 16S rRNA and rpoB genes were deposited in the publicly-accessible TreeBASE data repository with the project number 14166. Acknowledgments We thank Dr. Darshan Brar at IRRI for providing the rice material, Dr. Peter Kämpfer and Dr. Roger Stephan for providing the type strains of Enterobacter radicincitans, Enterobacter turicensis, Enterobacter helveticus and Enterobacter pulveris, and Dr. Jiří Jirout for assistance in the fatty acid analyses (BC ASCR, ISB). This study was supported by the joint RUG-WUR initiative on rice endophytes in the context of a DOE-JGI project on the rice endophyte metagenome and by a grant provided by the FWF (National Science Foundation, grant no. P 21261-B03) to A.S. P.R.H. was supported by the Soil Biotechnology Foundation. Electronic supplementary material Additional file 2: Figure S2: Maximum-likelihood tree based on rpoB gene sequences showing the phylogenetic position of Enterobacter oryziphilus sp. nov.