We have studied the influence of the thickness and the heat

treatment of the buffer layers and the deposition conditions of the BaTiO3 on the crystallinity, orientation, and morphology of the BaTiO3 films. Methods Buffer layer deposition Polyvinyl pyrrolidone (45% in water) dissolved in 2-propanol is spin-coated onto the silicon substrate as an adhesion layer prior to the buffer layer deposition. Buffer layer solutions are prepared by dissolving lanthanum nitrate hydrate in 2-propanol. The solution is spin-coated on the silicon wafers at 3,000 rpm for 45 s and subjected to a heat treatment at 450°C for 5 min. Lanthanum nitrate hydrate (La(NO3)3) decomposes through nine endothermic weight loss processes with increasing temperature [17]. Between 440°C and 570°C, the lanthanum nitrate hydrate Napabucasin is decomposed to the intermediate-phase lanthanum oxynitrate (LaONO3). The thickness of the obtained buffer layers in this work ranges between 6 and 10nm as measured with ellipsometry. BaTiO3 thin-film deposition Reagent

grade barium acetate Ba(CH3COO)2 and titanium butoxide Ti(C4H9O)4 are used as precursor materials for barium and titanium, and glacial acetic acid and 2-methoxy ethanol are used as the solvents. The molarity of the solution is 0.25 M. The BTO precursor sol is spin-coated at 3,500 rpm for 45 s, followed by pyrolysis on a hot stage at 350°C to burn out the organic components. This leads to a film thickness of about 30 nm. This process is repeated three or four times TSA HDAC mouse to obtain a film thickness

around 100 nm. Then, the silicon substrate with the BTO amorphous film is subjected to a high-temperature annealing at 600°C to 750°C for 20 min, with a tube annealing furnace in ambient air. The ramping rates for heating and cooling of the specimen in the annealing system are 100°C/min and −50°C/min, respectively. The process cycle (two or three spin coatings and subsequent high-temperature treatment) is repeated several times to obtain an oriented thin film with a thickness of a few 100 nm. X-ray diffraction measurements The samples are first this website cleaned with acetone, isopropanol, and de-ionized water. The measurements are carried out with a D8 Discover diffractometer (Bruker Technologies Ltd., Billerica, MA, USA) with CuKα radiation. The diffractograms are 2-hydroxyphytanoyl-CoA lyase recorded for 2θ angles between 15° and 64°, with a step size of 0.004° and time step of 1.2 s. Focused ion beam etching/scanning electron microscopy The cross-section images of the specimens are prepared by a FEI Nova 600 Nanolab dual-beam focused ion beam system (FIB; FEI Co., Hillsboro, OR, USA) and an associated scanning electron microscope (SEM). It allows simultaneous milling and imaging of the specimens. The SEM column is equipped with a high-performance field-emission gun electron source, whereas the FIB system has a gallium liquid metal ion source. Atomic force microscopy The surface roughness of the BTO thin films are measured by atomic force microscopy (AFM) analysis.

Zool Scr 2009,38(3):323–331.CrossRef 26. Fenchel T: Ecology of protozoa: the biology of free-living phagotrophic protists. Madison, Wisconsin: Science Tech Publishers; 1987. 27. Cawthorn RJ, Lynn DH, Despres B, MacMillan R, Maloney R, Loughlin M, Bayer R: Description of Anophryoides haemophila n. sp. (Scuticociliatida: Orchitophryidae), a pathogen of American lobsters Homarus americanus . Dis Aquat Org 1996, 24:143–148.CrossRef 28. Gomez-Saladin E, Small EB: Prey-induced transformation of Miamiensis avidus SCH727965 cell line strain Ma/2 by a soluble factor. J Eukaryot

Microbiol 1993, 40:550–556.CrossRef 29. Dunthorn M, Foissner W, Katz LA: Molecular phylogenetic analysis of class Colpodea (phylum Ciliophora) using broad taxon sampling. Mol Phylogenet Evol 2008, 46:316–327.PubMedCrossRef 30. Utz LRP, Eizirik E: Molecular phylogenetics of subclass Peritrichia (Ciliophora: Oligohymenophorea) based Saracatinib datasheet on expanded analysis of 18S rRNA sequences. J Eukaryot Microbiol 2007, 54:303–305.PubMedCrossRef 31. Carr M, Leadbeater BSC, Hassan R, Nelson M, Baldauf SL: Selleck ABT 263 Molecular phylogeny of choanoflagellates, the sister group of Metazoa. Proc Natl Acad Sci USA 2008,105(43):16641–16646.PubMedCrossRef 32. Hyman LH: The invertebrates: protozoa through Ctenophora. New York: McGraw-Hill; 1940. 33. King N, Westbrook MJ, Young SL, Kuo A, Abedin M, et al.: The genome of the choanoflagellate Monosiga brevicollis

and the origins of metazoan multicellularity. Nature 2008, 451:783–788.PubMedCrossRef 34. Rokas A: The origins of multicellularity and the early history of the genetic toolkit for animal development. Annu Rev Genet 2008, 42:235–251.PubMedCrossRef 35. Fauré-Fremiet E: Growth and differentiation of the colonies of Zoothamnium alternans (Clap. and Lachm.). Biol Bull 1930, 58:28–51.CrossRef 36. Summers FM: Some aspects of normal development in the colonial ciliate Zoothamnium

alternans . Biol Bull 1938, 74:117–129.CrossRef 37. Crowe SA, Jones C, Katsev GBA3 S, Magen C, O’ Neill AH, Sturm A, Canfield DE, Haffner GD, Mucci A, Sundby B, Fowle DA: Photoferrotrophs thrive in an Archean Ocean analogue. Proc Natl Acad Sci USA 2008,105(41):15938–15943.PubMedCrossRef 38. Zerkle AL, House CH, Brantley SL: Biogeochemical signatures through time as inferred from whole microbial genomes. Am J Sci 2005,305(6–8):467–502.CrossRef 39. Wilbert N: Eine verbesserte Technik der Protargolimprägnation für Ciliaten. Mikrokosmos 1975, 64:171–179. 40. Perez-Uz B, Guinea A: Morphology and infraciliature of a marine scuticociliate with a polymorphic life cycle: Urocryptum tortum n. gen., n. comb. J Eukaryot Microbiol 2001,48(3):338–347.PubMedCrossRef 41. Tompkins J, DeVille MM, Day JG, Turner MF: Culture collection of algae and protozoa. Catalogue of strains. Cumbria, UK: The Culture Collection of Algae and Protozoa; 1995. 42. Medlin L, Elwood HJ, Stickel S, Sogin ML: The characterization of enzymatically amplified eukaryotic 16S-like rRNA-coding regions.

This potential may be considered particularly large, when P515 (ECS) and “P515 flux” are measured simultaneously with other probes of photosynthetic electron transport, like CO2-uptake, O2-evolution, chlorophyll fluorescence, and P700. After calibration

of the flux GSI-IX ic50 signal by CO2-uptake or O2-evolution measurements, it may serve a non-invasive, continuously measured optical proxy of the in vivo rate of photosynthetic electron flow. Acknowledgments We thank Thomas Simon and Frank Reichel for skillful this website help in the development of the Dual-PAM-100, and Reinhold Fischer, Hardy Skiba, and Doris Steinert for their dedicated help with the instrumentation and set-up of the combined gas exchange measurements. Open AccessThis article is distributed under the terms of the Creative Commons Attribution License which permits any use, distribution, and reproduction in any medium, provided

the original author(s) and the source are credited. References Aronsson H, Schöttler MA, Kelly AA, Sundquist C, Dörmanns P, Karim S, Jarvis P (2008) Monogalactosyldiacylglycerol deficiency in Arabidopsis affects pigment composition in the prolamellar body and impairs thylakoid membrane energization and photoprotection in leaves. Plant Physiol 148:580–592PubMedCrossRef Asada K (1999) The water–water cycle in eFT-508 cost chloroplasts: 3-mercaptopyruvate sulfurtransferase scavenging of active oxygen and dissipation of excess photons. Annu Rev Plant Physiol Plant Mol Biol 50:601–639PubMedCrossRef Avenson TJ, Cruz JA, Kramer DM (2004a) Modulation of energy-dependent quenching of excitons (qE) in antenna of higher plants. Proc Natl Acad Sci USA 101:5530–5535PubMedCrossRef Avenson TJ, Cruz JA, Kanazawa A, Kramer DM (2004b) Regulating the proton budget of higher plant photosynthesis. Proc Natl Acad Sci USA 102:9709–9713CrossRef Avenson TJ, Kanazawa A, Cruz JA, Takizawa K, Ettinger WE, Kramer DM (2005a) Integrating the proton circuit into photosynthesis: progress and challenges.

Plant Cell Environ 28:97–109CrossRef Avenson TJ, Cruz JE, Kanazawa A, Kramer DM (2005b) Regulating the proton budget of higher plant photosynthesis. Proc Natl Acad Sci USA 102:9709–9713PubMedCrossRef Berry S, Rumberg B (1999) Proton to electron stoichiometry in electron transport of spinach thylakoids. Biochim Biophys Acta 1410:248–261 Bilger W, Heber U, Schreiber U (1988) Kinetic relationship between energy-dependent fluorescence quenching, light scattering, chlorophyll luminescence and proton pumping in intact leaves. Z Naturforsch 43c:877–887 Bilger W, Björkman O, Thayer SS (1989) Light-induced spectral absorbance changes in relation to photosynthesis and the epoxidation state of xanthophyll cycle components in cotton leaves.

At the time of hospital admission for surgical treatment of their hip fracture, hip fracture patients are reported to be malnourished, and

the nutritional AZD5363 cost status can deteriorate further during hospital admission because of a spontaneous reduction in food intake due to lack of appetite or nausea [4–9]. Malnutrition in hip fracture patients is reported to be associated with impaired muscle function, disability, loss of independency, decreased quality of life, delayed wound healing, higher complication rate, prolonged rehabilitation time, and increased mortality rate [7, 8, 10–17]. Both hip fracture patients and malnourished patients in general have MI-503 nmr an increased use of health care as compared with well-nourished and non-fracture patients, and it is expected that it would result in higher health care costs [18–21]. Early treatment of malnutrition is of vital importance to minimize losses and to achieve rapid weight recovery after hip fracture. In the past decades, several studies have been conducted to determine the effectiveness of nutritional supplementation on length of stay, postoperative complications, mortality, nutritional status and functional status. Furthermore, within the past decades, economic evaluations have gained more and more attention, and their importance has increased

because of the

continuous rising health care expenses and the limited Nutlin-3 cost budgets available. As a consequence, new or additional treatments should not only have to be effective but also cost-effective. MTMR9 Previous research on costs and cost-effectiveness of nutritional support or intervention is scarce. A few studies have shown that health care costs can be reduced by nutritional support in malnourished elderly [20, 22, 23]. Kruizenga et al. [24] reported that nutritional screening and treatment of malnourished patients at an early stage of hospitalization is cost-effective. Although several studies have shown the effectiveness of nutritional support in elderly hip fracture patients, none of these studies have incorporated an economic or cost-effectiveness evaluation. Therefore, the aim of the present study was to investigate the cost-effectiveness of an intensive dietary intervention comprising combined dietetic counseling and oral nutritional supplementation, as compared with usual nutritional care in elderly subjects after hip fracture from a societal perspective with a time horizon of 6 months. Methods Subjects Eligible were patients admitted for surgical treatment of hip fracture, aged ≥55 years [25]. Patients were excluded if they had a pathological or periprosthetic fracture; a disease of bone metabolism (e.g.

However, changes in cell morphology were not as evident as in other Gram negative bacteria. The majority of F. psychrophilum cells remaining as long and thin bacilli, few showing round enlargements, and in some cases, they adopted a ring-like conformation. The response of F. columnare to short- and long-term starvation has been studied based on cell culturability [8–10] but characterization on the morphological and physiological

changes that accompany this phenomenon have find more not been investigated in this species. The objective of this study was to assess the potential of F. columnare to survive under starvation conditions as well as to characterize the ultrastructural changes in cell morphology this website that accompanies this process. Methods Bacterial strains Four previously characterized F. columnare strains were used in this study representing two of the genomovars described learn more within the species [15, 16]. Genomovar I strains included the type strain ATCC

23463, originally isolated from Chinook salmon, and strain ARS-1 recovered from channel catfish. Genomovar II was represented by strains ALG-00-530 and AL-02-036 isolated from channel catfish and largemouth bass, respectively. Virulence between genomovar I and II strains is significantly different in channel catfish. Selected genomovar II strains are highly virulent in channel catfish fingerlings (mortality >90%) while genomovar I strains are less (ARS-1 produces <50% mortality) or not virulent (ATCC 23463) [17]. Bacteria were stored at −80°C as glycerol stocks and routinely

cultured on modified Shieh agar (MS) or broth with shaking (125 rpm) at 28±2°C for 24–48 h [18]. Survival under starvation conditions Individual colonies Ribonuclease T1 from each strain were inoculated into 4 ml of MS broth and incubated at 28±2°C overnight with shaking. Overnight cultures (4 ml) were inoculated into 36 ml of MS broth and incubated overnight as before. Cultures were centrifuged at 3000 g for 5 min, resuspended in 9 ml of ultrapure type I water (ThermoScientific Barnstead E-pure), stored in the dark at room temperature, and monitored for a period of two weeks. Three independent replicates per strain were conducted for statistical analysis. At day 1, day 7 and day 14, an aliquot from each of the 12 tubes (4 strains × 3 replicates) was taken for i) colony forming unit (CFU) counts, ii) light microscopy, and iii) scanning electron microscopy (SEM) (see below). Ultrastructural analysis Changes in morphology were monitored periodically using light microscopy, SEM, and transmission electron microscopy (TEM). For light microscopy, cells (5 μl of culture) were air dried on a microscope glass slide, stained with safranin and observed using a Leica DM2500 with differential interference contrast (Leica Microsystems, USA). For SEM, cells (5 μl of culture) were fixed in 2.5% glutaraldehyde (v/v) at 4°C overnight.

43, CI = 1.11–1.85) (Thun et al. 2006). Given that DNA adducts are associated with the development of lung tumors, it is plausible

that African Americans would have higher adduct levels (Tang et al. 2001; Peluso et al. 2005). However, our data do not support this hypothesis. There are some possible explanations for our findings. First, we measured adducts in a surrogate tissue (WBCs) rather than the target tissue (lung). Thus, the WBC DNA adducts may not represent the aggregate amount of tobacco-induced damage occurring in the lungs. Moreover, WBCs may represent a surrogate for www.selleckchem.com/products/cb-5083.html other exposures in adults that are not experienced by children, to the same Repotrectinib molecular weight extent. Thus, these exposures could be associated with a smoking lifestyle. In addition, our cohort consisted solely of non-smoking children; studies of racial differences in lung cancer have focused primarily on smoking adults, and may be racial differences in DNA adducts occur only among Selleck SB525334 active smokers. Lastly, the absence of racial differences in 1-Hydroxypyrene could

indicate that there may have been unmeasured sources of PACs in our study. Our results are subject to some limitations. First, our study was cross-sectional in design. At best, we could only identify an association between adducts and tobacco smoke exposure. Second, air nicotine levels were only measured in the main activity room G protein-coupled receptor kinase of the home. Thus, there may have been unmeasured exposures in other parts of the home or outside of the home that contributed to adduct formation. Thus, parents may have smoked around their child in other parts of the home that would not have been captured by the

nicotine dosimeter. In addition, we were unable to determine the impact of the air cleaners on PACs—compounds likely leading to adduct formation—as airborne levels of these compounds were not directly measured. Unfortunately, urine 1-HP levels cannot differentiate inhaled versus ingested exposure to PACs, and 1-HP levels reflect only recent exposure to PAC materials. While we did measure serum and hair cotinine levels that would capture ETS exposures outside of the home, it is well known that these biomarkers differ significantly by race. Still, we did not find any association of WBC DNA adducts with serum cotinine or hair cotinine—which operate as aggregate biomarkers of exposure. Third, we only measured PAC-DNA adducts, which may represent only a fraction of DNA damage induced by tobacco smoke. Aromatic amines are another family of compounds found in ETS that can form adducts with DNA (Talaska et al. 1991a, b; Hecht 2001, 2004). Fourth, there may have been sources of PACs other than ETS—such as exhaust from automobiles or dietary intake—that were not measured by the air nicotine dosimeters.

The samples were applied later on the hydrogenation of styrene (Figure 1) for a comparison with results from the commercially available activated carbon-supported Pd and Pt catalysts, Pt/C and Pd/C. Figure 1 The hydrogenation reaction of styrene to ethylbenzene and ethylcyclohexane. Methods The synthesis of GANT61 in vivo graphite oxide and graphene followed the well-known Hammer’s method [25]. A 250-mL round bottomed flask filled with 25 mL concentrated sulfuric acid (98%, Adrich, St. Blebbistatin Louis, MO, USA) was held in an iced bath. After 5 to 10 min, 10 mL fumed nitric acid was added slowly in 15 min. Then, graphite powder (1.0 g, with particle size <45 μm) was added into the mixture under vigorous stirring for

30 min with the flask held in the iced bath. Then 22 g potassium chlorate was added into the solution in 30 min, and the mixture was stirred at room temperature for 96 h. The solution was centrifuged with a suitable

amount (about 200 to 300 mL) of deionized (DI) water added under an iced bath temperature. Removal of liquid phase, followed by addition of DI water and then centrifugation, was repeated for three times. The mud-like residue was dried at 80°C for 12 h to produce the graphite oxide. The nanocomposite check details synthesis followed a procedure similar to that reported in our previous study [26]. Graphite oxide (250 mg) was added in 250 mL DI water and stirred for 30 min before addition of 1.4 g NaBH4, and the mixture was kept at 80°C for 1 h. Prior to sulfonation, the solution was centrifuged for collection of residues that were rinsed with methanol for three times then dried at 80°C under the N2 atmosphere for 1 h. The graphite oxide was sulfonated and exfoliated to graphene with the following procedure: in a 500-mL round-bottomed flask, the residues SDHB (158 mg) in 300 mL DI water were dispersed using an ultrasonic bath for 30 min. Separately,

sulfanilic acid (140 mg) and potassium nitrate (50 mg) were introduced into a 100-mL beaker containing DI water (40 mL) employing an iced bath. After being mixed well, the solution was added with 1 N HCl (1 mL) and then the solution was poured into the above mentioned round-bottomed flask and stirred for 2 h in the iced bath. Centrifugation followed by removal of aqueous solution resulted in the sulfonated graphene, which was rinsed with methanol for a few times then dried at 80°C under the N2 atmosphere. The microwave-assisted synthesis of Pt/GE and Pt/GO was performed using a CEM Discover Du7046 microwave set (Matthews, NC, USA) with 80 W power output for 30 s then held at 80°C for 5 min. The nanocomposites were prepared with sulfonated graphene or graphite oxide (100 mg) as substrates together with grinded K2PtCl6 at 14.5, 355, or 15 mg, respectively, plus 2-hydroxyethanaminium formate (5.0 g), in Pyrex glass tubes (results shown in Table 1).

So far, OSI-906 manufacturer Comparative tools for exploring the potential influences of species-specific PTMs on host-virus interactions have not been found. Here we develop a web-based

selleck chemical interactive database – CAPIH (Comparative Analysis of Protein Interactions for HIV-1) – for comparative studies of genetic differences between the human proteins involved in host-HIV protein interactions and their orthologues retrieved from three mammalian species: chimpanzee (Pan troglodyte), rhesus macaque (Macaca mulatta), and mouse (Mus musculus). The three latter species are all important animal models for HIV studies [15–17]. Understanding the differences in host-virus interplay between human and the model species is the basis for correct interpretation Selleck GS1101 of animal-based HIV studies. Furthermore, by comparing protein interactions between species, one can potentially identify key differences that underlie chimpanzee resistance to AIDS. To facilitate inter-species comparisons of host-HIV PPIs, four main functions are provided in CAPIH. Firstly, the interface shows the presence or absence of orthologous proteins, thus enabling users to pinpoint missing protein components in the host-HIV interaction network.

Secondly, the multiple sequence alignments of orthologous proteins enable users to identify species-specific amino acid substitutions, nucleotide substitutions, and indels. This information is helpful for inferring functional changes of orthologous proteins. Thirdly, predictions of 7 types of species-only PTMs (phosphorylation, methylation, sumoylation, acetylation, sulfation, N-glycosylation, and O-glycosylation) for each HIV-interacting host protein PAK5 are presented for analyses of potential PTM influences on protein interactions and signal/regulatory pathway. We also collect experimentally verified PTMs in human proteins. Fourthly, CAPIH shows potential PPI hot sites on the multiple sequence alignments. Through the visualized interface, researchers can easily spot multiple host factors that directly or indirectly interact

with the same HIV protein, and consider how changes in one member protein may affect the protein interaction network. Construction and content CAPIH organization and implementation The data compiling process is illustrated in Figure 1A. We retrieved a total of 1,447 HIV-1 interacting human proteins from the HIV-1, Human Protein Interaction Database [18] (the November 13, 2007 freeze). The human-chimpanzee-macaque-mouse orthologous proteins were downloaded from the Ensembl genome browser (release 47), which were identified by the Ensembl project using the Markov clustering algorithm [19]. Note that not all the retrieved human proteins have orthologues in all of the three compared species. In the cases of one-to-many/many-to-many orthologous relationships, only the protein pairs with the reciprocally highest similarity were selected. All of the protein and nucleotide sequences were downloaded from Ensembl.

C cortex, PL photobiont layer, Pho photobiont, M medulla, Hy fungal hyphae ROS generation, chlorophyll autofluorescence and lipid peroxidation during lichen rehydration Although several works

have described an extracellular oxidative burst during rehydration in some lichen species, virtually nothing is known about intracellular ROS production and its relationship to abiotic stress. In order to determine whether intracellular ROS release follows the rehydration of R. farinacea thalli, 10 μM of the fluorescent probe DFCH2-DA was added to the deionized water https://www.selleckchem.com/products/Trichostatin-A.html used for rehydration. The samples were observed by fluorescence and confocal microscopy 3-4 h after rehydration. The presence of 2′,7′- dichlorofluorescin (DCF), the fluorescent oxidation product of DCFH2, indicated the intracellular production of free radicals during lichen rehydration

(Figure 2B-D). DCF was especially concentrated in the lichen cortex. No significant green autofluorescence was detected in the absence of the probe (Figure 2A). Confocal microscopy showed discrete points of green fluorescence around several large photobionts (Figure 2E), probably due to mycobiont hyphae tips. Figure 2 ROS 3-Methyladenine molecular weight in rehydrated R. farinacea thalli. Thalli of R. farinacea rehydrated with deionized water and 10 μM DCFH2-DA and observed 3-4 h post-rehydration. A, B, C, D ROS content, as revealed by the green fluorescence emission of DCF under a fluorescence microscope (magnification: 400× for A, B and 1000× for C, D); E overlay of confocal microscopy images reveals ROS distribution around some of the photobionts (green fluorescence); F overlay of confocal microscopy images of ROS content of R. farinacea thalli that had

been rehydrated with c-PTIO 200 μM, arrows point to photobionts this website photobleached by the confocal laser during the observation (oxPho). Red fluorescence is due to the photobiont’s chlorophyll in all cases. Each micrograph is representative of several images corresponding to independent click here samples. C cortex, M medulla, PL photobiont layer, Pho photobiont, oxPho photobleached photobiont, Hy fungal hyphae A fluorometric kinetics of intracellular free radical production in Ramalina farinacea thalli was performed in order to confirm microscopical data. Figure 3A demonstrates that the rate of intracellular free radical production in recently rehydrated thalli was much higher than the rate of intracellular free radical production in thalli kept in the hydrated state during the previous 24 h. Furthermore, intracellular release of free radicals during rehydration under physiological conditions was biphasic with an initial exponential phase of 20 min followed by a linear phase (Figure 3B). Chlorophyll autofluorescence was simultaneously recorded since this parameter is a surrogate of the levels and integrity of this molecule and therefore of the photosynthetic status of the cell.

2-kb PCR product carrying dndB with

introduced NdeI and BamHI sites (with C-terminal His-tag) was amplified and Etomoxir concentration cloned into pMD18-T to give pJTU68. Then the corresponding NdeI-BamHI DNA fragment from Batimastat mouse pJTU68 was introduced into pHZ1272 between the restriction sites NdeI and BamHI to give pJTU81. dndC expression vector: using pHZ1904 as template, and wlr7 and wlr11 as primers, a 1.5-kb PCR product carrying dndC with introduced NdeI and BamHI sites (with C-terminal His-tag) was amplified and cloned into pMD18-T to give pJTU72. Then dndC from pJTU72 was introduced into pHZ1272 between the restriction sites NdeI and BamHI to give pJTU86. dndD expression vector: using pHZ1904 as template, and dnd-1 and dnd-2 as primers, a 2.0-kb PCR product carrying dndD with introduced NdeI and BamHI sites was amplified, digested with the corresponding enzymes and cloned into pET15b to generate pHZ2893. Then dndD from pHZ2893 was introduced into pHZ1272 between the restriction sites NdeI and BamHI to give pJTU64. dndE expression vector: using pHZ1904 as template, and dndE-L and dndE-R as primers, a 0.4-kb PCR product carrying dndE with introduced NdeI site was amplified and cloned into pMD18-T to give pJTU180. Then dndE from pJTU180 was introduced into pHZ1272 after digestion with NdeI and BamHI to EPZ015666 chemical structure give pJTU65. Over-expression and purification of DndD protein After IPTG induction, E. coli BL21 (DE3) containing

pHZ2893 over-expressed the DndD fusion protein with a His-tag at the N-terminal end. The fusion protein as inclusion bodies was further purified with an ÄKTA-fast protein liquid chromatography system (FPLC) (Amersham Pharmacia Biotech) and a 5-ml HiTrap chelating column (Amersham Pharmacia Biotech) under denaturing condition. The fusion protein was used for the production of rabbit anti-DndD polyclonal antibody. RT-PCR analysis of dnd genes Carnitine palmitoyltransferase II RNA extraction was according to the standard protocol of RNeasy Protect Bacteria Midi Kit from Qiagen Co. Ltd. RT-PCR experiments were performed according to the standard protocol of OneStep RT-PCR Kit from the same company. Primers are listed in Table 1. Acknowledgements We are very grateful to Prof. Sir David Hopwood, FRS for his continuous support

and encouragement throughout this study for many years, and help for the editing of the manuscript. The authors wish to thank the National Science Foundation of China (NSFC), the Ministry of Science and Technology 973 and 863 programs, the Ministry of Education of China, the Shanghai Municipal Council of Science and Technology and Shanghai Leading Academic Discipline Project for research supports. Electronic supplementary material Additional file 1: Additional table 1.Table displaying bacterial strains and plasmids. (DOC 56 KB) References 1. Hattman S: Unusual modification of bacteriophage Mu DNA. J Virol 1979,32(2):468–475.PubMed 2. Hattman S: Specificity of the bacteriophage Mu mom+ -controlled DNA modification. J Virol 1980,34(1):277–279.PubMed 3.