Plants of the genus Mentha produce a class of natural products kn

Plants of the genus Mentha produce a class of natural products known as mono-terpenes (C10), characterized by p-menthone skeleton. Members of this genus are the only sources for the production of one of the most economically important essential oil, menthol, throughout the world [12]. Mentha piperita, commonly called peppermint, is a well-known herbal remedy used for a variety of symptoms and diseases, recognized for its carminative, stimulating, antispasmodic, antiseptic, antibacterial, and antifungal activities

[4, 13, 14]. However, their use for clinical purposes is limited by the high volatility of the major compounds. Talazoparib Due to their high biocompatibility [15] and superparamagnetic behavior, magnetite nanoparticles (Fe3O4) have attracted attention to their potential applications especially

in biomedical fields [16, 17], such as magnetic resonance imaging [18–20], hyperthermia [21], biomedical separation and purification [22], bone cancer treatment [21], inhibition of biofilm development [23, 24], stabilization of volatile organic compounds [25], antitumoral treatment without application of any alternating magnetic field [26], drug delivery or targeting [27–33], modular microfluidic system for magnetic-responsive controlled drug release, and cell culture [34].This paper reports a new nano-modified prosthetic device surface with anti-pathogenic properties based on magnetite nanoparticles and M. piperita essential oil. Methods Materials All chemicals were used as received. FeCl3 (99.99%), FeSO4·7H2O (99.00%), NH3(28% NH3 in H2O, Tamoxifen research buy ≥99.99% trace metal basis), lauric acid (C12) (98.00%), CHCl3 (anhydrous, ≥99%, contains 0.5% to 1.0% ethanol as stabilizer),and CH3OH (anhydrous, 99.8%) were purchased from Sigma-Aldrich. Prosthetic device represented by catheter sections were obtained from ENT (Otolarincology), Department of Coltea Hospital, Bucharest, Romania. Axenfeld syndrome Fabrication of nano-modified prosthetic device For the fabrication of the nano-modified prosthetic device, we used a recently published

method [35] in order to design a new anti-pathogenic surface coated with nanofluid by combining the unique properties of magnetite nanoparticles to prevent biofilm development and the antimicrobial activity of M. piperita essential oil. M. piperita plant material was purchased from a local supplier and subjected to essential oil extraction. A Neo Clevenger-type apparatus was used to perform microwave-assisted extractions. Chemical composition was settled by GC-MS analysis according to our recently published paper [36]. Magnetite (Fe3O4) is usually prepared by precipitation method [37–39]. The core/shell nanostructure used in this paper was prepared and characterized using a method we previously described [40].

PBP2a detection was performed using monoclonal PBP2a antibody (1:

PBP2a detection was performed using monoclonal PBP2a antibody (1:20000) from the MRSA-screen kit (Denka Seiken). Acknowledgements We would like to thank Frances O’Brien (School of Biomedical Sciences, Curtin University of Technology) for determining the MLST types of strains ZH44 and ZH73. We would also like to thank Sibylle Burger for technical assistance and Dr. P. Hunziker, of the Functional Genomics Centre

Zurich, University of Zurich, for protein analysis. We are also grateful to T. Bae (Department of Microbiology, University of Chicago) for providing the plasmid pKOR1. This study was supported by the Swiss National Science Foundation grant NF31-117707/1. References 1. Kirby WMM: Extraction of a highly potent penicillin inactivator from penicillin resistant styaphylococci. Science 1944,99(2579):452–453.CrossRefPubMed 2. Hartman Tyrosine Kinase Inhibitor Library clinical trial BJ, Tomasz A: Low-affinity penicillin-binding protein Deforolimus ic50 associated with beta-lactam resistance in Staphylococcus aureus. J Bacteriol 1984,158(2):513–516.PubMed 3. Reynolds PE, Brown FJ: Penicillin-binding proteins of β-lactam-resistant strains of Staphylococcus aureus . Effect of growth conditions. FEBS Lett 1985,192(1):28–32.CrossRefPubMed 4. Lim D, Strynadka NC: Structural basis for the β-lactam resistance of PBP2a from methicillin-resistant Staphylococcus aureus. Nat Struct Biol 2002,9(11):870–876.PubMed 5. Sharma VK, Hackbarth CJ, Dickinson

TM, Archer GL: Interaction of native and mutant MecI repressors with sequences that regulate mecA , the gene encoding penicillin binding protein 2a in methicillin-resistant staphylococci. J Bacteriol 1998,180(8):2160–2166.PubMed 6. Gregory PD, Lewis RA, Curnock SP, Dyke KG: Studies of the repressor (BlaI) of beta-lactamase synthesis in Staphylococcus aureus. Mol Microbiol 1997,24(5):1025–1037.CrossRefPubMed 7. Zhang HZ, Hackbarth CJ, Chansky KM, Chambers HF: A

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PLoS One 2011, 6:e19235 PubMedCentralPubMedCrossRef 75 Bignell D

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factor may be involved in Streptomyces coelicolor antibiotic production and sporulation. Microbiol 2000, 146:2161–2173. 76. Westbye AB, Leung MM, Florizone SM, Taylor TA, Johnson JA, Fogg PC, Beatty JT: Phosphate concentration and the putative sensor kinase protein CckA modulate cell lysis and release of the Rhodobacter capsulatus gene transfer agent. J Bacteriol 2013, 195:5025–5040.PubMedCrossRef Competing interests The authors declare that they have no competing interests. Authors’ contributions RGM and ASL designed the research. RGM performed the experiments and analyzed the data. RGM

and ASL wrote the manuscript. Both authors read and approved the final manuscript.”
“Background Zymomonas mobilis is a Gram-negative facultative anaerobic bacterium, which has attracted significant interest over recent years for its use in the industrial-scale production of ‘bioethanol’ [1–5]. This microorganism is able to ferment glucose, fructose or sucrose to ethanol, with extremely high molecular efficiencies and minimum accompanying levels of biomass formation. As a ‘generally regarded as safe’ (GRAS) microorganism, Z. mobilis has also been used for a variety of other biotechnological purposes, such as the production of levan (polyfructan) [6, 7] or amino acids [8]. Over the past 20 years or so, significant effort has been Selleckchem AZD4547 spent on genetically ‘engineering’ its metabolic capabilities and physiological TCL activities. These have largely focused on extending its limited substrate range, enabling it to utilize carbohydrates that are abundant in lignocellulosic feedstocks [2, 4, 5, 9–12]. Genetic engineering applications in Z. mobilis have commonly utilized plasmid vectors housing heterologous genes encoding proteins with the desired functionalities [12]. Cloning vectors that are routinely used in Escherichia coli, such as those derived from pBR322 or pUC18, cannot be stably-maintained

in Z. mobilis[12]. On the other hand, several types of bacterial broad-host range plasmids are able to replicate in Z. mobilis cells (e.g. derivatives of pBBR1MCS, RSF1010 and the incW R plasmid Sa), and have been used for a variety of heterologous gene expression applications. However, they are prone to structural (genetic) instability, and their relatively large size constrains gene cloning strategies [12–15]. Consequently, the most common approach for heterologous gene expression in Z. mobilis has involved E. coli – Z. mobilis shuttle vectors; which incorporate replicons from E. coli plasmids, as well as those from native plasmids isolated from various Z. mobilis strains [12, 13, 16–22]. Four native plasmids from Z.

For the PC measurements, the incident light, namely, the infrared

For the PC measurements, the incident light, namely, the infrared (IR) beam from the FTIR spectrometer, was perpendicular to the mesa upper surface; and for our structure on the mesa upper surface, the area exposed to the light occupies about 75% of the total area. Results and discussion Figure 1a gives the scheme of selleckchem one unit of coupled QDs lasing layers in one period.

Figure 1b shows the atomic force microscopy (AFM) image of one-period QDCL with another unit of coupled QDs lasing layers (indicated by the dashed rectangle in Figure 1a) on top. The average diameter of QDs is about 30 nm, with a height of 2.5 nm. The entire structural quality of the QDCL wafer was confirmed by the X-ray diffraction (XRD) spectrum as shown in Figure 1c. In the XRD simulation, we treated the QD layer as a two-dimensional InAs layer with a homogeneous thickness corresponding to the nominal deposit amount, which was

strained biaxially to match the lattice constant of InP. The experimental zeroth peak shows a nearly perfect lattice match to the InP substrate, which demonstrates that the active region layers have been properly strain-balanced to give a net zero strain. The accurate match of the simulated curve and the experimental curve shows an extremely good control Dinaciclib over the growth parameters across the entire 30-period layer sequences. The cross-sectional view of transmission electron microscopy (TEM) images of a portion of the 30-period QDCL shown in Figure 2a,b gives the direct and clear evidences of distinct coupled QDs layers in the active core. What is more, the X-ray energy dispersion spectra (EDS) result obtained along cross section line of coupled QDs layers gives indium contents at different points. The ‘star’ represents the discrete data point of X-ray energy dispersion spectrum at each position along cross section line (Figure 2b) of coupled QDs layers of the TEM sample. Based on the finite scattered experimental Miconazole data points, we sketch the continuous curve of indium composition along cross

section line with periodic oscillation characteristic. The periodic oscillation characteristic of indium relative contents as shown in Figure 2c gives the additional evidence of QDs in the active region. This result is consistent with the AFM one. Figure 2 TEM image and EDS results. (a) TEM image of a portion of the cleaved cross section of a QDCL active region. (b) The enlargement image of a portion of Figure 2a for clarity, and the white line gives a clear indication of QDs distribution parallel to the growth layer. (c) Indium relative content along the indicated white line in Figure 2b measured by X-ray energy dispersion spectra. A schematic conduction band diagram of one period of the active layers is shown in Figure 3a. The design computation is based on 1D Schroedinger equation of envelope function approximation from the point of view of simplicity.

Applied Environmental Microbiology 1989, 55:1957–1962 33 George

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Appl Microbiol 2010, 51:645–649 PubMedCrossRef 30 v


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Figure 7 shows that the genes of the urease gene cluster are tran

Figure 7 shows that the genes of the urease gene cluster are transcribed as a single transcript. Control assays confirmed that the purified RNA was free of contaminating DNA (Figure 7, lanes b). Figure 7 Reverse transcriptase PCR of urease operon. Ethidium bromide stained agarose gel showing results of reverse transcriptase PCR with H. influenzae strain 11P6H. Lanes: a) purified RNA with reverse transcriptase and Taq DNA polymerase; b) purified RNA with Taq DNA polymerase (negative control); c) purified DNA with Taq DNA polymerase (positive

control). Lane H2O is a water control. Oligonucleotide Selleckchem NVP-LDE225 primers were designed to span adjacent genes in the gene cluster as noted at the top of the gel (See Table 1). Molecular size markers are noted in base pairs on the right. Presence of urease operon in clinical

isolates To determine whether the urease operon is present in clinical isolates of H influenzae, 20 clinical isolates, including 10 otitis media strains and 10 COPD strains were studied by PCR. Primers corresponding to genes located in the 5′ region (ureA), central region (ureC) and 3′ region (ureH) of the Roxadustat ic50 operon were designed. Amplicons of identical size were obtained from 20 of 20 clinical isolates with all 3 sets of primers (Figure 8). These results indicate that the urease operon is present in all strains tested and that no variation was observed in the lengths of these genes in diverse strains tested. Figure 8 Urease operon in clinical isolates. Ethidium bromide stained agarose gels showing amplicons of genes in the urease gene cluster as noted on the left. Lanes a through j, amplicons from COPD sputum

isolates 6P8H1, 14P14H1, 24P17H1, 27P5H1, 33P18H1, 43P2H1, 55P3H1, 66P33H1, 74P16H1, 91P18H1, respectively. Lanes k through t, middle ear fluid aspirate isolates 1749, 1826, 6699, 6700, 4R, 17R, 26R, 47R, P86, P113, respectively. Molecular size standards are noted on the right in kilobases. A BLASTn search with the sequence corresponding to the urease operon was performed to determine which strains of H. influenzae whose genomes are available in ZD1839 ic50 GenBank contained the urease operon. Five of 6 strains whose complete genome has been sequenced contain the urease operon. A high degree of sequence similarity in the urease operon is present among the 5 strains. In strain R2866, which is urease negative, the urease operon is replaced by a single gene with homology to the gonococcal mtrF gene [40]. Sequence analysis of the same region of 9 additional urease negative strains revealed sequence that is very similar to that of strain R2866 [40]. Transcription of the ureC during growth in pooled human sputum To assess expression of urease in conditions that simulate conditions in the human respiratory tract in COPD, transcription of ureC was measured in H.

iniae HD-1 s

iniae HD-1 LY2109761 manufacturer using rabbit anti-MtsA antibodies (Figure 5A). MtsA was detected in the particulate fraction of the cells when the cellular fractions were prepared by centrifugation of the crude cell lysate (the first treatment). MtsA was found to be associated with the protoplast and cell wall extracts when the cellular fractions were prepared by protoplast

formation. After separation of the protoplasts, MtsA was detected in the particulate fraction (the second treatment). Figure 5 Detection of the subcellular localization of MtsA in S. iniae HD-1 by western blotting. (A) The cellular fractions of S. iniae HD-1 and rabbit anti-MtsA antibodies were used for the western-blot assay. Lane 1, S. iniae HD-1 FDA approved Drug Library price lysate; lane 2, soluble fraction of cells; lane 3, particulate fraction of cells; lane 4, cell wall extracts; lane 5, protoplast; lane 6, particulate fraction of protoplasts; and lane 7, soluble fraction of protoplasts. (B) Surface exposure of MtsA. Cells (lanes 1 and 2), cell wall extracts (lanes 3 and 4), and protoplasts (lanes 5 and 6) of S. iniae HD-1 were treated with proteinase K and analyzed by western blotting. Lanes 1, 3 and 5 show the untreated control, while lanes 2, 4 and 6 show samples treated with proteinase K for 1 h. To detect surface exposure of MtsA, cells of S. iniae

HD-1 cells were harvested, washed, centrifuged, and resuspended in PBS. The cells were subjected to proteinase K (5 μg ml-1) treatment with gentle agitation selleck kinase inhibitor at room temperature for 1 h, and the cells were collected. Western blotting showed that peptide fragments in the cells can be detected after 1 h incubation with proteinase K. However, when the cell wall

extracts and protoplasts were used in the experiment, it were completely hydrolyzed and no peptide fragments were detected (Figure 5B). Together, this result indicated that MtsA is not exposure on surface, but is on the outside of the cytoplasmic membrane and is buried inside the cell wall. MtsA had heme-binding activity To examine whether heme is the chromophore associated with MtsA, the pyridine hemochrome assay was performed [28]. The UV-visible absorption spectrum of purified MtsA exhibited peaks at 275, 420, 525, and 560 nm, which were identical to those obtained from purified KatG, a well-known heme-containing protein with spectral peaks at 418, 524, and 556 nm. The molar ratio of associated heme to purified MtsA was 0.806 (Figure 6), this value is consistent with the hypothesis that one protein molecule is associated with one heme molecule. Figure 6 Detection of the heme-binding activity of purified MtsA by the pyridine hemochrome assay. (A) UV-visible absorption spectrum of 20 μM purified MtsA (■ line) in 50 mM Tris-HCl (pH 8.0). (B) UV-visible absorption spectrum of 20 μM purified KatG (Δ line) in 50 mM Tris-HCl (pH 8.0).

In 2M + LB nutrient medium, these mutants had reduced levels of t

In 2M + LB nutrient medium, these mutants had reduced levels of the maltoporin (band 2) and the presence of the putative porin (band 4) protein in replacement of the OmpU-like porin (band 5) compared to the wild-type (Figure 5C). Expression of a single gene cassette in trans maintains normal growth after generation of strains with

deleted cassettes Since mutant d16-60 (cassettes 16 to 60 deleted) had normal growth phenotypes compared to the wild-type, at least one cassette gene located between cassettes 7 and 16 has a strong pleiotropic affect. All eight cassettes within this region, except cassette 11, encode small hypothetical proteins with homology only to other cassette proteins. Therefore, nothing could be inferred regarding their putative function. However, cassette 11 includes a gene, encoding a 257 amino acid protein with pfam selleck screening library http://​pfam.​sanger.​ac.​uk/​ identifying

two domains; 1) an uncharacterized NERD domain at residues 31-150 that has weak homology to nucleases and is commonly associated with other protein domains involved in DNA processing [22], 2) a DNA-binding C4-zinc finger domain at residues 216-257 found in topoisomerase I proteins and involved in removing excessive negative supercoils from DNA [23]. Based on this bioinformatics analysis one possible biochemical function of the cassette 11 gene product is as a DNA topoisomerase. In addition, experiments with a mutated topoisomerase I (topA) gene have described phenotypes that are similar to those observed in the d8-60 mutants. Most notably, in characterized topA CHIR-99021 molecular weight mutants, this includes the requirement for a compensatory mutation, emergence of spontaneous mutants and alterations in the composition of outermembrane porin proteins [23–28]. To test Quisqualic acid for the cassette 11 gene product being responsible for the phenotype of the mutants described above, the plasmid pMAQ1082 was constructed which comprises only this cassette gene cloned into the vector pJAK16 (Methods). pMAQ1082 was then transformed into the merodiploid strain MD7. MD7 has a complete

DAT722 cassette array and is the strain that was used to create the original deletion mutants (Methods and Figure 1). MD7/pMAQ1082 possesses a phenotype identical to that of DAT722 with respect to porin profiles and growth in LB20 and 2M media. From this strain, a deletion mutant was created, DAT722Δ/pMAQ1082 with the same cassettes deleted as strains d8-60a, b and c. The strain DAT722Δ/pMAQ1082 had no major growth defect (Figure 6) and possessed a wild type outermembrane protein profile in all tested media (Figure 5D, E, F). A slight decrease in growth rate was observed in 2M + pyruvate (Figure 6), which may be explained by the up-regulation of a protein (Figure 4F; marked with an asterisk) that is likely due to cassette 11 being removed from its native promoter.

Invest New Drugs 2011, 29:239–247 PubMedCrossRef 95 Wang Q, Zhen

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