Antimicrobial susceptibility testing Susceptibility to a standard panel of antimicrobial agents http://​www.​danmap.​org was determined by microbroth dilution and interpreted according to Clinical and Laboratory Standards Institute guidelines [32], except for ciprofloxacin (≥0.125 μg/mL was used as breakpoint). Phage typing Phage typing was performed by the National Food Institute, Technical University of Denmark according to the phage typing scheme developed by Callow [33] and extended by Anderson et al. [34]. Strains that react with phages, but have a profile not included in the phage selleck compound typing scheme, are denoted Reaction Does Not Conform (RDNC). DNA microarray The DNA microarray used

in this study was previously described [35]. A set of 281 gene-specific

57-60 mer oligonucleotide probes were designed using the program Array Designer 4.1 (Premier Biosoft, Palo Alto, CA, USA). The oligonucleotides were spotted on glass slides using a QArray Mini Arrayer (Genetix, New Milton, UK). Hybridized spots were visualized in a GenePix Selumetinib 4000B laser scanner (Axon, Foster City, CA). Each oligonucleotide allows the detection of the presence or absence of a characteristic sequence previously described in Salmonella. The microarray used gives no information on the location of a gene or target sequence and can only score its presence or absence. Uncertain array results were resolved by PCR using primers described previously [35]. Data analysis Analysis of the DNA microarray data was performed as previously described [35]. A comparison was made by importing array values, gene present or absent, into BioNumerics 5.1 (Applied Maths, Sint-Martens-Latem, Belgium) as character data. An Unweighted Pair Group Method with Arithmetic mean (UPGMA) dendrogram (Fig. 2) was calculated by simple matching of binary coefficients on the basis of a geneset consisting of all genes from the array except the serotyping markers and the resistance markers. Multiple-locus

variable-number of tandem-repeat analysis (MLVA) MLVA was performed as described previously [36] to ensure that the strains were likely to be epidemiologically unrelated. The MLVA repeats were calculated selleck inhibitor and named according to the method described recently [37]. Pulsed-field gel electrophoresis (PFGE) PFGE was carried out with XbaI restriction enzyme according to the Pulse-Net protocol [38], gels were analyzed in BioNumerics 5.1, and profiles were assigned by comparison to a database with known Danish profiles (see additional file 1: Xba I PFGE profiles of all isolates). Sequence typing Multilocus sequence typing (MLST) was carried out as previously described [39] and the alleles and the sequence types were assigned according to the MLST scheme on http://​mlst.​ucc.​ie/​mlst/​mlst/​dbs/​Senterica/​. Acknowledgements EL was partly funded by the Danish Research Council. We are grateful to the patients without whose kind participation this study would not have been possible.

Matullo G, Palli D, Peluso M, Guarrera S, Carturan S, Celentano E, Krogh V, Munnia A, Tumino R, Polidoro S, Piazza A, Vineis P: XRCC1, XRCC3, XPD gene polymorphisms, smoking and (32)P-DNA adducts in a sample of healthy subjects. Carcinogenesis 2001, 22: 1437–1445.CrossRefPubMed 22. Pachkowski

BF, Winkel S, Kubota Y, Swenberg JA, Millikan RC, Nakamura J: XRCC1 genotype and breast cancer: functional studies and epidemiologic data show interactions between XRCC1 codon 280 His and smoking. Cancer Res 2006, 66: 2860–2868.CrossRefPubMed 23. Butkiewicz D, Rusin M, Enewold L, Shields PG, Chorazy Cabozantinib solubility dmso M, Harris CC: Genetic polymorphisms in DNA repair genes and risk of lung cancer. Carcinogenesis 2001, 22: 593–597.CrossRefPubMed 24. Sancar A: Excision repair in mammalian cells. J Biol Chem 1995, 270: 15915–15918.PubMed 25. Benhamou S, Sarasin A: ERCC2/XPD gene polymorphisms and lung cancer: a HuGE review. Am ZD1839 order J Epidemiol 2005, 161: 1–14.CrossRefPubMed 26. Qiao Y, Spitz MR, Shen H, Guo Z, Shete S, Hedayati M, Grossman L, Mohrenweiser H, Wei Q: Modulation of repair of ultraviolet damage in the host-cell reactivation assay by polymorphic XPC and XPD/ERCC2 genotypes. Carcinogenesis 2002, 23: 295–299.CrossRefPubMed 27. Spitz MR, Wu X, Wang Y, Wang LE,

Shete S, Amos CI, Guo Z, Lei L, Mohrenweiser H, Wei Q: Modulation of nucleotide excision repair capacity by XPD polymorphisms in lung cancer patients. Cancer Res 2001, 61: 1354–1357.PubMed 28. Au WW, Salama SA, Sierra-Torres CH: Functional characterization of polymorphisms in DNA repair genes using cytogenetic challenge assays. Environ Health Perspect 2003, 111: 1843–1850.CrossRefPubMed 29. Au WW, Navasumrit P, Ruchirawat M: Use of biomarkers to characterize functions of polymorphic DNA repair genotypes. Int J Hyg Environ Health 2004, 207: 301–313.CrossRefPubMed 30. Costa S, Pinto D, Pereira

D, Vasconcelos A, fonso-Lopes C, Osorio T, Lopes C, Medeiros R: Importance of xeroderma pigmentosum group D polymorphisms in susceptibility to ovarian cancer. Cancer Lett 2007, 246: 324–330.CrossRefPubMed 31. Lunn RM, Helzlsouer KJ, Parshad R, Umbach DM, Harris EL, Sanford KK, Bell DA: XPD polymorphisms: effects on DNA repair proficiency. Carcinogenesis 2000, 21: 551–555.CrossRefPubMed 32. Seker H, Butkiewicz D, Bowman ED, Rusin M, Hedayati from M, Grossman L, Harris CC: Functional significance of XPD polymorphic variants: attenuated apoptosis in human lymphoblastoid cells with the XPD 312 Asp/Asp genotype. Cancer Res 2001, 61: 7430–7434.PubMed 33. Wei Q, Cheng L, Amos CI, Wang LE, Guo Z, Hong WK, Spitz MR: Repair of tobacco carcinogen-induced DNA adducts and lung cancer risk: a molecular epidemiologic study. J Natl Cancer Inst 2000, 92: 1764–1772.CrossRefPubMed 34. Benhamou S, Sarasin A: ERCC2/XPD gene polymorphisms and cancer risk. Mutagenesis 2002, 17: 463–469.CrossRefPubMed 35. Caggana M, Kilgallen J, Conroy JM, Wiencke JK, Kelsey KT, Miike R, Chen P, Wrensch MR: Associations between ERCC2 polymorphisms and gliomas.

According to [17], the appearance of these two low-temperature phases of Ni silicides after annealing in vacuum would be evidence that the original Ni film has been completely (or nearly completely) consumed by the growing Ni2Si phase).a In this case, the volume fraction of Ni2Si/NiSi 85:15 (taking into account all uncertainties, the maximum estimate yields 100% of Ni2Si); the mass fraction of Ni2Si exceeds 88%. This obviously contradicts our TEM observations and makes us assume the presence of the heaviest of the Ni silicides,

Ni3Si [18], which also may form at low temperatures, especially taking into account the possible presence of oxygen in the metal film that, according to [17, 22], impedes diffusion of Ni atoms to VX-765 molecular weight LY2157299 mouse Ni/Ni2Si interface and, in our opinion, may result in simultaneous formation of Ni2Si and Ni3Si phases in the silicide film. If our assumption is true, the silicide film might be composed,

by a rough estimate, of 20% to 40% of Ni3Si, 30% to 60% of Ni2Si, and 10% to 30% of NiSi in respective proportions to give a total of 100% of the silicide film volume. The lightest (the least dense) silicide phase having a Si-rich stoichiometry (disilicide) may also be available in the form of a thin diffusion layer at the Ni silicide/poly-Si interface (this does not contradict our observations) [23]; it may affect the barrier height of the whole silicide layer, however [20]. I-V characteristics of the structures (Figure 2a,b) with low-resistance

poly-Si ( ), which forms in our process, manifest a diode behavior with the rectification ratios changing from about 100 to about 20 for the temperature varied from 22°C to 70°C (Figure Lenvatinib order 2c). At liquid nitrogen temperature, the rectification becomes more pronounced and exceeds 1,000 at biases exceeding 2 V (Figure 3). It should be noticed that at forward bias, the negative lead was set on the silicide top contact pad, whereas the positive one was set on the contact pad to the polysilicon film. Figure 2 I – V characteristics of the Ni silicide/poly-Si structure and its rectification ratios at different temperatures. (a) Forward and (b) reverse biases; (c) rectification ratio vs. the applied voltage. Figure 3 I – V characteristics of the Ni silicide/poly-Si structure and its rectification ratios at liquid nitrogen and room temperatures. (a) I-V characteristics and (b) rectification ratio as a function of the applied bias. Photo-electromotive force (emf) spectra obtained at 300 and 80 K (Figure 4) demonstrate photoresponse for photons with energies greater and less than the Si bandgap width (E g) as well as the presence of a number of potential barriers in the diode film. Room temperature measurements with and without a silicon filter have revealed the only barrier with the height Φ rt≈0.

The little discrepancy between these two spectra might have originated from the resonant excitation of Er3+. Besides, the peak around 3.8 eV which appears in the PLE spectra might be related to the optical excitation of the Si NCs since the introduction of the Si NCs would enhance the PL intensity of both Si=O states and Er3+. Conclusions In summary, the efficient luminecence of Er3+ in the SROEr film is achieved by the energy transfer process from fast recombination centers Alpelisib price (LCs). The SROEr films with abundant LCs (WOBs, NOVs, and Si=O states) and Si NCs are prepared by electron beam evaporation following a post-annealing process. Intense

and stable PL of LCs dominated by the Si=O states is obtained in the SROEr matrix. From the investigation of the evolution of the PL properties and AG-014699 mouse microstructures from the SROEr films, we consider the fast energy transfer from the Si=O states to Er3+ as the main transfer mechanism. The introduction of the Si NCs induces the Si=O states and facilitates the photon absorption of the

Si=O states, which is essential to obtain intense PL from both Si=O states and Er3+. Further improvement of the PL property of both the Si=O states and Er3+ might be achieved by optimizing the annealing condition of the SROEr films. Authors’ information DL received his Ph.D. degree in the State Key Laboratory of Silicon Materials and Department of Material

Science and Engineering from Zhejiang University, Hangzhou, China, in 2002. He is currently an Associate Professor Protirelin in the Department of Material Science and Engineering at Zhejiang University. His current research interests include the synthesis of plasmonic microstructure, application of plasmonic microstructure on solar cells, Raman and luminescence, and silicon photonics. LJ, LX, and FW are currently Ph.D. students in the State Key Laboratory of Silicon Materials and Department of Materials Science and Engineering, Zhejiang University, Hangzhou, China. Their current research interests include luminescence from erbium-doped silicon-rich oxide matrix, silicon-rich nitride matrix, and dislocations in silicon, silicon nitride-based light-emitting devices, and localized surface plasmon resonance of metal nanostructures. DY received his B.S. degree from Zhejiang University, Hangzhou, China, in 1985, and Ph.D. degree in Semiconductor Materials from the State Key Laboratory of Silicon Materials in Zhejiang University, Hangzhou, China, in 1991. He has been with the Institute of Metal Materials in Tohoku University, Japan, and worked for Freiberg University, Germany, from 1995 to 1997. He is currently the director of the State Key Laboratory of Silicon Materials.

The ubiquitous expression of a ‘humanized’ Cdh1 in this mouse allows the investigation of InlA-Cdh1 and InlB-Met interactions in vivo. We have previously taken a different route to generate an InlA and InlB permissive L. monocytogenes mouse infection model through an approach we call pathogen ‘murinisation’ [12]. Based on structural information on the recognition complex of InlA with the N-terminal

domain of Cdh1, two amino acids in InlA were replaced (Ser192Asn and Tyr369Ser), dramatically increasing the binding affinity of murine Cdh1 to InlA [12]. By introducing these two mutations into the listerial inlA locus, a variant strain of L. monocytogenes EGD-e (Lmo-InlAm) was generated which was able to cross the murine intestinal barrier and to

induce symptoms of listeriosis click here after oral inoculation [12]. In contrast to the Cdh1 transgenic mouse models, this mouse model permits the analysis of orally acquired listeriosis without the need to cross in ‘humanized’ alleles of Cdh1. In this study, we have employed a previously generated bioluminescent L. monocytogenes EGD-e strain (Lmo-InlA-mur-lux) ‘murinised’ for the two Ser192Asn and Tyr369Ser inlA mutations [17] and a ‘non-murinised’, isogenic control strain (Lmo-EGD-lux) to analyse host responses after oral infection in four different inbred strains of mice. C3HeB/FeJ, A/J, BALB/cJ, and C57BL/6J mice were intragastrically inoculated with Lmo-InlA-mur-lux and Lmo-EGD-lux and bacterial

mTOR inhibitor dissemination to internal organs was analysed using bioluminescent in vivo imaging (BLI). These mouse inbred strains were chosen for the C1GALT1 study as they represent priority strains for the mouse phenome project [18] and their degree of host resistance to oral L. monocytogenes infection has never been investigated and compared in a single study under identical infection challenge conditions. We report here that infection with murinised Listeria resulted in earlier onset of listeriosis compared to infections with the non-murinised Listeria strain in different mouse genetic backgrounds. BLI enabled accurate measurement of bacterial dissemination over consecutive days in the acute stage of disease and showed that Lmo-InlA-mur-lux disseminated earlier from the intestine to target organs in the C3HeB/FeJ, A/J, and BALB/cJ mice. However, no increase in dissemination to the brain was detected, revealing that Listeria uses different mechanisms to cross the intestinal epithelium and to cross the blood–brain barrier. Results Dynamics of Lmo-InlA-mur-lux and Lmo-EGD-lux dissemination visualized by BLI To compare the dissemination dynamics of the murinised and wildtype L.

aureus. (iii) Increased sensitivity to UV irradiation and mitomycin C, a phenotype in agreement with a role of RecU in DNA damage repair. (iv) Increased recruitment of the DNA translocase SpoIIIE. In B. subtilis, RecU has been shown to bias homologous recombination towards non-crossover

products [7, 11], decreasing the formation of chromosome dimers that would not be properly segregated into the daughter cells [46–48]. When present, chromosome dimers can be resolved by dedicated recombinases in a process that requires the presence of at least one of the two DNA translocases, SpoIIIE or SftA [49]. Furthermore, the presence of septal SpoIIIE foci was proposed to be associated with its role in post-septational chromosome partitioning Rapamycin ic50 [38]. Therefore, the fact that approximately half of the S. aureus cells grown in the absence of RecU had SpoIIIE-YFP foci (compared to 10% of the cells grown in its presence), suggests that RecU has a major role in chromosome segregation, maybe through biasing recombination towards non-crossover

products. (v) The presence of septa placed over the DNA, a phenotype that could be caused by segregation defects or, alternatively, by the lack of a cell division checkpoint required to prevent septum formation over the DNA (see below). Together, the phenotypes observed for RecU depleted cells strongly point to an important role of this protein in DNA repair and chromosome segregation, in agreement with what would be expected for a Holliday junction resolvase. In the course of S. aureus cell division, the synthesis of cell wall occurs Selleckchem VX 809 at the septum, which progressively closes to originate the two daughter cells. During this process the chromosome is replicated and the two resulting DNA molecules are segregated. Tight coordination between chromosome segregation (which requires

RecU) and septum synthesis (which requires PBP2, encoded in the same triclocarban operon as RecU), two biosynthetically unrelated events, is therefore essential for proper division, to ensure that the septum does not form over the nucleoid, which would result in DNA damage. Given that the genetic organization of the recU-pbp2 operon is maintained in other gram-positive bacteria [19, 21, 22], we hypothesized that co-regulation of the expression of these two proteins could be central for the coordination of cell division events. We have abolished this co-regulation (but maintained the presence of RecU in the cell) in strain 8325-4recUi by placing an inducible copy of recU in the distant spa locus, under the control of the P spac promoter and deleting the native gene from the recU-pbp2 operon. When this mutant is incubated with IPTG, RecU is produced from the ectopic spa locus while PBP2 is expressed from its native locus, under the control of its native promoters.

Aspergillus-specific

IgG antibodies in the sera of all patients were determined by an indirect ELISA using filtrate proteins of A. fumigatus (1 μg/ml) as the coating antigen (sera diluted 1:1000). All sera were stored at -70°C. Sera of IA patients and controls were pooled separately for immunoproteomics analysis. According to EORTC-MSG criteria, proven IA refers to histopathologic evidence of tissue invasion by septated, acutely-branching filamentous fungi, together with Deforolimus a positive culture (sputum and/or bronchoalveolar lavage) [39]. The study protocol was approved by the Ethics Committee of the hospital and informed consent was obtained from all patients included in the study. Preparation of extracellular proteins A. fumigatus (strain CMCC (f) A1a) was obtained from the Microbial Culture Collection Management Committee of China, Medical Mycology 17-AAG concentration Center. The fungus was first grown on Sabouraud agar plates at 37°C for 3 days. The conidia were collected and incubated in yeast-extract-peptone-glucose (YEPG) broth (1% yeast extract, 2% peptone, and 2% glucose) in a 500-ml

flask on a shaker at 37°C for 14 days. Then, the culture supernatant was collected by filtration. The proteins were recovered by trichloroacetic acid (TCA) precipitation, as described previously [40]. Finally, the precipitates were resuspended in two-dimensional electrophoresis (2-DE; 7 M urea, 2 M thiourea, 4% [w/v] CHAPS, 1% [w/v] DTT, 1% protease inhibitor cocktail [v/v], and 2% [v/v] IPG buffer [pH 3-10]) lysis buffer, and stored at -70°C. The protein concentration was determined by the Bradford method using BSA as the standard. Two-dimensional electrophoresis and Western blot analysis Samples Flucloronide containing

150 μg of filtrate protein were separated by 2-DE, as described elsewhere [41], using immobilized, non-linear pH 3-10 gradient strips (24 cm; Amersham Biosciences, Uppsala, Sweden) for isoelectric focusing, and 12.5% sodium dodecylsulfate polyacrylamide gels for the second dimension separation. All gels were silver-stained according to published procedures [42] or electrotransferred to polyvinylidene fluoride (PVDF) membranes [43]. Three replicates were run for each sample. Western blot was performed as described previously [44]. Briefly, the membranes were probed with primary antibody (pooled sera of patients with proven IA and pooled control sera [1:1000 dilution in each case]) at 4°C overnight. Subsequently, the membranes were thrice washed with Tris-buffered saline (pH 7.5) containing 0.05% (v/v) Tween-20 (TBST) for 10 min and incubated with horseradish peroxidase (HRP)-conjugated goat anti-human IgG (1:2000 dilution) for 2 h at room temperature. The membranes were then washed with TBST and the signal was detected with an enhanced chemiluminescence detection kit (Amersham Biosciences, Uppsala, Sweden).

Nanoscale Res Lett 2011, 6:438–442.CrossRef 25. Bhattacharjee B, Ganguli D, Iakoubovskii K, Stesmans A, Chaudhuri S: Synthesis and characterization of sol–gel derived ZnS: Mn 2+ nanocrystallites embedded in a silica matrix. Bull Mater Sci 2001, 25:175–180.CrossRef 26. Wang L, Dai J, Liu X, Zhu Z, Huang X, Wu P: Morphology-controlling synthesis of ZnS through a hydrothermal/solvthermal Palbociclib datasheet method. Ceram Int 2010, 38:1873–1878.CrossRef 27. Amaranatha Reddy D, Murali G, Vijayalakshmi RP, Reddy BK, Sreedhar B: Effect of Cr doping on the structural and optical properties of ZnS nanoparticles. Cryst Res Technol 2011, 46:73–736.CrossRef 28. Poornaprakash B, Amaranatha Reddy

D, Murali G, Madhusudhana Rao N, Vijayalakshmi RP, Reddy https://www.selleckchem.com/products/Metformin-hydrochloride(Glucophage).html BK: Composition dependent room temperature ferromagnetism and PL intensity of cobalt doped ZnS nanoparticles. J Alloys Compd 2013, 577:79–85.CrossRef 29. Amaranatha Reddy D, Murali G, Poornaprakash B, Vijayalakshmi RP, Reddy BK: Structural, optical and magnetic properties of Zn 0.97− x Cu x Cr 0.03 S nanoparticles. Appl Surf Sci 2012, 258:5206–5211.CrossRef 30. Pal M, Mathews NR, Morales ER, Jimenez JM, G y , Mathew X: Synthesis of Eu +3 -doped ZnS nanoparticles

by a wet chemical route and its characterization. Opt Mater 2013, 35:2664–2669.CrossRef 31. Hu H, Zhang W: Synthesis and properties of transition metals and rare-earth metals doped ZnS nanoparticles. Opt Mater 2006, 28:536–550.CrossRef 32. Yang P, Lu M, Xu D, Yuan D, Zhou G: ZnS nanocrystals co-activated by transition metals and rare-earthmetals-a new class of luminescent materials. J Lumin 2001, 93:101–105.CrossRef 33. Iqbal MJ, Iqbal S: Synthesis of stable and highly luminescent beryllium and magnesium doped ZnS quantum dots suitable for design of photonic and sensor material. J Lumin 2013, 134:739–746.CrossRef 34. Chen Z, Li XX, Chen N, Du G, Li Y, Liu G, Suen AYM: Study on

the optical properties of Zn 1− x Mg x S (0 ≤  x  ≤ 0.55) quantum dots prepared by precipitation Pyruvate dehydrogenase lipoamide kinase isozyme 1 method. Mater Sci Eng B 2012, 177:337–340.CrossRef 35. Pathak CS, Mishra DD, Agarwal V, Mandal MK: Blue light emission from barium doped zinc sulfide nanoparticles. Ceram Int 2012, 38:5497–5500.CrossRef 36. Shi Q, Wang Z, Liu Y, Yang B, Wang G, Wang W, Zhang J: Single-phased emission-tunable Mg-doped ZnO phosphors for white LEDs. J Alloys Compd 2013, 553:172–176.CrossRef 37. Vinodkumar E, Roshith R, Kumar V: Mg-doped ZnO nanoparticles for efficient sunlight-driven photocatalysis. Appl Mater Interfaces 2012, 4:2717–2725.CrossRef 38. Justin Raj C, Karthick SN, Hemalatha KV, Son MK, Kim HJ, Prabakar K: Magnesium doped ZnO nanoparticles embedded ZnO nanorod hybrid electrodes for dye sensitized solar cells. J Sol–gel Sci Technol 2012, 62:453–459.CrossRef 39. Jin C, Park S, Kim H, Soyeon A, Lee C: CO Gas-sensor based on Pt-functionalized Mg-doped ZnO nanowires.

Using high concentrations of hydrogen in the staining procedure has the advantage that Hyd-3 activity is detectable after a few minutes’ exposure, while Hyd-2 is not detectable under these conditions, possibly due to the low abundance of the enzyme in extracts of E. coli coupled with the brief exposure to hydrogen. Hyd-3, like Hyd-1, is a more abundant

enzyme and this possibly explains the rapid visualization of both these enzymes after only 10 min exposure to high hydrogen concentrations. selleck The fact that the FHL complex is active in H2 oxidation contrasts the physiological direction of the reaction in the E. coli cell. This, therefore, might be an explanation for the comparatively high H2 concentrations required to drive the reaction in the direction of hydrogen oxidation. The similar redox potentials of formate and hydrogen do, however, indicate that this reaction should be freely reversible, possibly pointing to a role of a progenitor of the FHL complex in CO2 fixation [44]. Another possible explanation for the effect of hydrogen concentration on Hyd-3 activity is that high hydrogen concentrations drive the redox potential of a solution to more negative E h values [10]. For example

a 100% hydrogen atmosphere will result in a E h = -420 mV in anaerobic cultures, while a 5% hydrogen concentration in the headspace equates to a redox potential of around -370 mV and Anti-infection Compound Library a dissolved hydrogen concentration in cultures of maximally 40 μM at 25°C [36]. Our recent studies have shown that the [Fe-S]-cluster-containing small subunit of the hydrogenase must be associated with the large subunit in order for hydrogen-dependent BV reduction to occur [20]. It is possible that BV receives electrons from a [Fe-S] cluster. If this is the case, then hydrogen-dependent BV reduction by a component of Hyd-3 also possibly occurs via a [Fe-S] cluster; however, due to the considerable number of [Fe-S] cluster-containing subunits in the complex (HycB, HycF, HycG and the Fdh-H enzyme itself [20, 45]) future studies will PtdIns(3,4)P2 be required to elucidate whether BV can interact with one or several

sites in the complex. The use of the electron acceptor NBT enabled a clear distinction between Hyd-1 and Hyd-2 activities. Previous experiments have shown that PMS/NBT staining is sometimes non-specific due to interaction with protein-bound sulfhydryl groups and even BSA was shown to be capable of staining gels incubated with PMS/NBT [46]. We could clearly show in this study, however, that, of the hydrogenases in E. coli, only Hyd-1 was capable of the specific, hydrogen-dependent reduction of PMS/NBT. Notably, both respiratory Fdhs also showed a strong NBT-reducing activity, which correlates well with previous findings for these enzymes [21]. Hyd-1 is similar to the oxygen-tolerant hydrogenases of R. eutropha and it is equipped with two supernumerary cysteinyl residues, which coordinate the proximal [4Fe-3S]-cluster [9, 47].

6 mW

and the integration time 20 s. In each sample, we measured 10 points to obtain average Raman intensity as the reference used in the SERS enhancement factor calculation. The Raman peaks fitted from the baseline-removed Raman spectra using a Guassian-Lorentzian lineshape. Calculation of SERS enhancement factors We calculated the SERS enhancement factors of the single R6G molecule absorbed on our 3D nanostructures by the equation [3, 4, 12] (1) where EF was the enhancement factor, I SERS and I bulk are the Raman signal SAHA HDAC nmr intensities at 1,365 cm-1 band, which is a characteristic representative vibration wave number of R6G molecules adsorbed on the 3D nanostructure and from the bulk R6G, respectively; N surf and N bulk are the numbers of the R6G molecules absorbed on the 3D nanostructures and the bulk R6G molecules exposed to the laser spot, respectively. Results and discussion

The 3D nanostructural quartz substrates for SERS enhancement were fabricated by NSL. In detail, monolayer hcp-packed PS nanospheres were coated on the quartz substrate by self-assembly. Consequently, the PS-coated quartz substrate was precisely tailored by O2 plasma in a RIE system after removing solvents, using a recipe as radio frequency (RF) power of 40 W, pure O2 gas flow of 40 sccm, and chamber pressure of 2 Pa. We found that the lateral and vertical etching rates of PS nanosphere under this condition were both 300 nm/min. Such high lateral BTK inhibitor etching rate was suitable to tailor PS nanosphere, while for etching PS nanosphere, the O2 gas flow should be changed to 5 sccm so that the lateral etching rate

can be lowered to 10 nm/min and the vertical etching rate as 30 nm/min. Figure 1 illustrates the results after tailoring PS nanosphere under above recipe with different operating time. Figure 1a is a typical SEM image of hcp-packed PS nanosphere without tailoring and the inset picture is its cross-sectional SEM image; both of them demonstrated that the sample was a monolayer PS nanosphere dispersed on quartz substrate. With the O2 plasma operating time increased from 3 to 5 and 10 s, and the gaps between two adjacent nanospheres were increased from 10 nm to 25 and 37 nm, as shown in Figure 1b,c,d, respectively. Figure 1d also illustrates substantially that the top morphologies were bleary after 10-s RIE treatment. Branched chain aminotransferase Since PS nanosphere contacted with the quartz substrate only at one point, the whole sphere was etched through gaps. The geometry of the etched nanosphere is a crucial factor for the followed substrate etching to achieve 3D nanostructures. Figure 1 SEM images of PS nanospheres on quartz substrate. (a) Top morphology after self-assembly, and after O2 plasma tailoring with a typical gas pressure of 2 Pa, and O2 gas flow of 40 sccm, a radio-frequency (RF) power of 40 W, with the treatment time as (b) 3, (c) 5, and (d) 10 s, respectively. The quartz substrate was directly nanopatterned by RIE. The tailored PS nanosphere performed as the sacrificial mask.