Therefore, the generated mutant can be readily screened on an aga

Therefore, the generated mutant can be readily screened on an agar plate containing sucrose. Figure 1 Plasmids constructed to introduce an unmarked mutation into a large gene of non-competent bacteria. (A, B) Multiple cloning VS-4718 cell line sites (MCS) of pJQ200sk and pK18mob were substituted with that of pLOI2224, generating pJQFRT and pKFRT, respectively. The pJQFRT plasmid contains a single FRT site adjacent to a multiple

cloning site; p15A origin, a replication origin of E. coli; oriT, origin of transfer; SacB, a counter-selection marker; and GmR, a gentamicin resistance marker. The arrows indicate the primers used in PCR to amplify the substitute MCS. The nucleotide sequences of these primers are shown in Table 2. (C) A cassette containing tetR-Ptet

promoter and flp recombinase amplified by PCR from pFT-A was ligated with the inverse-PCR product of pKFRT. The resultant pKFRT/FLP plasmid contains a single FRT site adjacent to a multiple cloning site; TetR-FLP, flp recombinase gene under the control of the tetR regulation system; KmR, a kanamycin resistance marker; oriT, origin of transfer; and ColE1 origin, a replication origin of E. coli. Figure 2 Scheme for the unmarked deletion of a large gene by FLP/FRT recombination. The plasmid pJQFRT with the insertion of the upstream region of the target Autophagy inhibitor gene is integrated into the host chromosome by homologous recombination. Next, the plasmid pKFRT/FLP with the insertion of the downstream region of the target gene is integrated into the host chromosome by homologous recombination. As a result, the target gene is sandwiched between the two integrated plasmids. The expression of flp is induced by adding anhydrotetracycline, and then the target region is excised together with the integrated plasmids bracketed by the two FRT sites, leaving a single FRT site. In our methodology, the new gene replacement plasmids pJQFRT and pKFRT/FLP are used for introducing

the unmarked mutation. Since these plasmids are mobilized by bacterial conjugation, there is no concern about the nucleolytic degradation of the introduced plasmid DNA, unlike linear DNA. Loperamide Besides, flp recombinase is cloned under the regulation of the tet promoter in pKFRT/FLP and is integrated into the chromosome of the recipient strain after homologous recombination. Therefore, our method obviates the need for helper plasmids expressing FLP recombinase and λ Red recombinase, which prevents degradation of the introduced linear DNA [30]. Our method can be used in various species of Gram-negative bacteria except for E. coli and some enterobacteria, independent of their competency and recombination ability. Implementation of the new method for the deletion of a large gene from the Acinetobacter sp.

Therefore, sliding means for 20 adjacent dots were calculated and

Therefore, sliding means for 20 adjacent dots were calculated and plotted to help visualise patterns (red

dots, Figure 5). Again no general relationship between position along one axis and position along the other could be established. Nevertheless the ori and right loci appeared this website to behave similarly and the NS-right locus tended to be closer than ori and right to the cell centre. The ter locus was more peripheral than other loci in cells with a single focus (red dots). The same analysis was performed for the ori and ter loci after Ndd treatment (Figure 5). For the ter locus, distributions of the two cell classes were combined since they were not significantly different (Additional file 1, Figure S4D). In both cases, the sliding mean was consistent with the peripheral location of the loci. Equivalent patterns were obtained for the right and NS-right

loci in Ndd-treated cells (not shown). Foci located in the 0-0.1 cell length slice were more central than the other foci. This cell length slice corresponds VRT752271 molecular weight to the cell poles, where the membrane curvature modifies the cell width distribution of foci. This effect was detected only in Ndd-treated cells due to the enrichment of loci in this cell slice compared to control cells (Additional file 1, Figure S4C). Figure 5 Analysis of correlation of the position of foci along the cell length with that along the cell diameter. Graphs show the positions of foci of four loci in wt and Ndd-treated cells, as indicated in each panel, along the cell diameter (Y-axis) as a function of their position along the cell length (X-axis). The grey dots are individual foci. The red dots are sliding means of twenty adjacent foci (with a step of one focus). For the ori, right and NS-right loci in Ndd-untreated cells and for the ori and ter loci in Ndd-treated cells, the data from the different cell classes were combined, as these dataset do not statistically differ (see Figure 2). In the case of the ter locus in Ndd-untreated cells, only the data from cells with a single

focus are plotted. The dotted lines show the mean position of foci calculated from the 90% central model. Discussion We report that it is possible to assess the mean position of chromosome loci across the width of a rod-shaped bacterium using two-dimensional Protirelin pictures. We recorded the apparent position of fluorescence-tagged chromosomal loci along the diameter of a large number of cells and compared the resulting distributions to simulated distributions calculated from different positioning models. We analysed five loci mapping in four different chromosomal regions that behave differently during the cell cycle. For these five loci, we detected three different patterns, showing that our method can detect differences in cell width localisation. The ori and right loci appeared randomly distributed through a cell volume corresponding to the nucleoid, whereas the NS-right locus was more central and ter loci more peripheral.

1 (ATCC TIB-67TM) cell lines were maintained in Dulbecco’s Modifi

1 (ATCC TIB-67TM) cell lines were maintained in Dulbecco’s Modified Eagle Medium(DMEM), and Human Lung Carcinoma, A-549 cells (ATCC CCL-185TM) were maintained in Ham’s F-12 K medium (F-12 K) supplemented

with 10% Fetal bovine serum (FBS) at 37°C with 5% CO2. Cytotoxicity assays Bacteria were cultured in SS media overnight and were then sub-cultured in SS media to an optical density of ~0.5 at 600 nm. For cytotoxicity assays, bacteria were added to previously seeded cell monolayers in 12- or 24-well tissue CRT0066101 price culture plates at the indicated MOIs. The plates were centrifuged for 5 min at 60 x g and incubated for up to 4 h at 37°C with 5% CO2. To measure cell cytotoxicity, Lactate dehydrogenase (LDH) release was used as a surrogate marker for cell death. LDH release in the supernatant media was assayed using a CytoTox 96® non-radioactive cytotoxicity assay kit (Promega, Madison, WI), according to the manufacturer’s instructions. selleck compound The maximal LDH release was defined as 100% and was determined by adding lysis solution to uninfected monolayers, determining the absorbance at 490 nm,

and then subtracting the background value. Each sample was measured in triplicate in at least three independent experiments. Animal infection experiments Wild-type female C57BL/6NCr (B6) mice, 4–6 weeks of age, were purchased from Charles River Breeding Laboratories (Wilmington, MA). The animals were lightly sedated with isoflurane (Novation Laboratories, TX) prior to intranasal infection with the indicated number of CFU of bacteria in a total volume of 40 μl of phosphate-buffered

saline (PBS, Mediatech Inc, VA). Bacteria were cultured in SS media overnight and were then sub-cultured in SS media to an optical density of ~0.5 at 600 nm. Inocula were confirmed by plating serial dilutions. For survival curves, groups of four mice were inoculated with the indicated dose, and the percent survival was monitored over a 30-day period. Mice with lethal bordetellosis, indicated by ruffled fur, labored breathing, and diminished responsiveness, were euthanized to alleviate unnecessary suffering [24]. To enumerate the number of bacteria in respiratory organs, groups of three to four mice were sacrificed at the indicated time points and bacterial numbers in the lungs and tracheas were Oxymatrine quantified by plating dilutions of tissue homogenates on BG plates with appropriate antibiotics, following incubation at 37°C for 2 days. The mean ± the standard error was determined for each group. The statistical significance between different groups was calculated by Student’s two-tailed t-test. A significance level was set at P values of ≤0.05. All animal experiments were repeated at least three times with similar results. Murine survival percentage was analyzed with the Log-Rank (Mantel-Cox) test. All mice were maintained in UCLA animal research facilities according to National Institutes of Health and University of California Institutional Animal Care Committee guidelines.

The two characteristics of gluQ-rs described in this work, co-tra

The two characteristics of gluQ-rs described in this work, co-transcription with the upstream gene and the presence of a terminator immediately upstream, allow us to propose that both the transcription

and translation process could be regulated in the gluQ-rs gene. It has been described, that the presence of terminators upstream of the coding region might be part of a regulatory system such as a riboswitch [35]. Riboswitches for genes involved in queuosine formation have been described, in which the precursor preQ1 is the ligand of the mRNA structure [36]. Using the riboswitch server ( [37]), we did not identify any potential riboswitch (data not shown). However we cannot discount that the terminator described here might be part of a regulatory circuit RAD001 in vitro similar to STA-9090 supplier a riboswitch, or that an unidentified protein might bind the terminator structure. GluQ modification and codon bias tRNA modifications present at the anticodon loop might be important for the accuracy of codon reading during the translation processes [13]. Morris et al., 1999 [14] proposed, based on molecular

modeling, that the tRNAAspQ34 might improve recognition of both GAC and GAU codons, consequently the interaction of the codon GAU with the anticodon of tRNAAspG34 could be less efficient. In fact, in S. flexneri there are a few genes such as sitA, virF and proX (an inducible gene under osmotic stress) that have a bias toward those codons that favor the Farnesyltransferase modified tRNA. Thus, while there is no obvious loss of plaque formation in the gluQ-rs mutant (data not shown), the absence of GluQ-RS may influence the expression of proteins such as SitA that are required for fitness of Shigella in the host [38]. Conclusions In this work we have shown that the expression of gluQ-rs, a gene codifying an enzyme involved in the formation of GluQ present on the tRNAAsp, is under the control of the dksA promoter. Also, we show the presence of a functional terminator that controls the expression of gluQ-rs. Finally, we present data that suggest that the presence of modification of the tRNAAsp is important for survival of the human pathogen Shigella flexneri under osmotic stress conditions. Methods Bacterial growth conditions

The bacterial strains and plasmids used in this study are described in Table 1. E. coli strains were maintained on LB-agar (10 g of tryptone per liter, 5 g of yeast extract per liter, 10 g of NaCl per liter and 15 g of agar per liter), Shigella strains were maintained on Trypticase Soy Agar plus 0.01% congo red. All strains were stored at −80°C in LB broth plus 20% glycerol. The bacteria were grown in LB broth adjusted to pH 7.4 with 40 mM MOPS (Merck) or M9 minimal media [24]. When necessary, ampicillin was added to a final concentration of 100 μg/ml. Bacterial growth was monitored by optical density at 600 nm (OD600). Bioinformatics tools to construct the phylogenetic tree The protein sequences were obtained from the Uniprot database (http://​www.

amazonensis (GenBank acc no EF559263); Lm,

amazonensis (GenBank acc. no. EF559263); Lm, selleck screening library L. major (TrEMBL acc. no. Q4QDR7); Li, L. infantum (GenBank acc. no. XP_001464939.1); Lb, L. brasiliensis (GenBank acc. no. XP_001564056.1); Tc, Trypanosoma cruzi (GenBank acc. no. XP_819954.1); Tb, Trypanosoma brucei (GenBank acc. no. AY910010); h, human (hTRF1 GenBank Acc. no. P54274.2; hTRF2 GenBank acc. no. Q15554). Figure 1 LaTRF is a homologue of mammalian and T. brucei telomeric TRFs.

(top) Position of the TRFH and Myb domains in LaTRF, according to rpsblast and bl2seq sequence analysis with T. brucei TRF. (bottom) ClustalW multiple alignment of the Myb-like DNA binding domains of human (hTRF2 and hTRF1), L. amazonensis (LaTRF), T. brucei (TbTRF) and T. cruzi (TcTRF) TRFs. In addition, like TbTRF, LaTRF shared sequence similarities with the canonical Myb-like domain and with the TRFH dimerization domain of human TRF1 and TRF2 (Fig 1-bottom and Table 1), but no sequence similarities were found with any other telobox CHIR-99021 in vitro protein (data not shown). Together, these results indicate that although LaTRF shares high sequence similarity with TbTRF, probably because the two species are phylogenetically related [26], further studies are required

to confer any functions to the Leishmania TRF homologue identified here. LaTRF is a nuclear protein that co-localizes with L. amazonensis telomeres In exponentially growing L. amazonensis promastigotes, LaTRF was detected only in nuclear protein extracts. A single ~82.5 kDa protein band was 3-mercaptopyruvate sulfurtransferase detected using anti-LaTRF serum (Fig 2 – top panel: lane 1). No protein was detected in cytoplasmic and total protein extracts (Fig 2 – top panel: lanes 2 and 3), indicating that LaTRF is a nuclear protein with very low intracellular abundance. As a control, Western blots were revealed with anti-LaRPA-1 serum, which recognizes a ~51.2 kDa telomeric protein band [23] (Fig 2 – bottom panel: lane 1) and also its phosphorylated forms (Fig 2 – bottom panel: lane 2; da Silveira & Cano, unpublished data). Figure 2 Expression of LaTRF in L. amazonensis promastigotes

extracts. Western blot analyses of extracts from 107 promastigotes/lane, grown in mid-log phase, were probed with anti-LaTRF serum (top panel) and anti-LaRPA-1 serum [31] as the loading control (bottom panel). Lane 1 – total protein extract (T), lane 2 – nuclear extract (N), lane 3 – cytoplasmic extract (C). We also developed an immunofluorescence assay combined with FISH, using anti-LaTRF serum and a PNA-telomere probe specific for TTAGGG repeats. As shown in Fig 3 (panels p1-4, merged images a and b), LaTRF is a nuclear protein that partially co-localizes with parasites telomeres, since some of the LaTRF signal coincided with telomeric foci and some did not (Fig 3, panels p1-4). In most cells, LaTRF appears as a diffuse signal spread all over the nucleoplasm and only in some cases it forms large punctuated foci, which seems to co-localize with the telomeric DNA (yellow dots in Fig 3, panels p2 and p4).

BC finished the characterization of CNTs and GNRs LC finished th

BC finished the characterization of CNTs and GNRs. LC finished the surface modification of MWNTs and GNRs. DM and FH finished the RGD conjugation with the surface of GNRs. WK and CD finished the result analysis. FH and WC finished the draft. LQ and CD finished the experiment design and manuscript revision. All authors of this paper have read and approved the final manuscript.”
“Background Enhancement of optical signals (Raman scattering, VX-680 cell line infrared absorption (IR), and luminescence) from

molecules adsorbed on the surface of nanostructured metals was considered in many papers published recently. The nanostructured gold, platinum, silver, copper, and other metals were used for the achievement of the enhancement effect. The enhancement

factor could achieve 106 for Raman scattering and 103 for IR absorption and luminescence [1, 2]. Moreover, surface-enhanced Raman scattering (SERS) effect allowed registration of the signal from a single molecule adsorbed on the nanostructured surface [3]. The mechanism of this effect possesses dual electromagnetic (EM) and chemical (CM) nature and is the matter of debate in the literature [1–4]. Earlier, we have registered enhancement in Raman and IR spectra SBE-��-CD supplier of different biomolecules adsorbed on carbon nanostructures: single-wall carbon nanotubes (SWCNTs) and graphene nanoflakes [5–7]. The maximum enhancement factor for Raman scattering of such nucleobases as thymine and adenine adsorbed on SWCNT was 10. It could be up to 80 on graphene oxide (GO) [8]. It is known from the literature that graphene could be used as enhancing support with enhancement factor from 17 to 69 [9–11]. The coherent anti-Stokes Raman scattering (CARS) technique is rather complex [12–14], and we found only a few papers devoted to its application for studying biomolecules [15–18]. The enhancement of CARS signal for molecules localized on nanostructured gold surface with an enhancement factor of approximately 105 was published in [17]. It was also established that this method is attractive for visualization of macromolecules medroxyprogesterone and cell components [19]. In the present paper, we used CARS to study

different carbon nanostructured materials (highly oriented pyrolytic graphite (HOPG), multiwall carbon nanotubes (MWCNTs), graphene nanoplatelets (GNPs), and GO) as well as the surface-enhanced coherent anti-Stokes Raman scattering (SECARS) effect for thymine (Thy) adsorbed on GO. Methods Samples Thy was purchased from Sigma-Aldrich (St. Louis, MO, USA) and used as received. The MWCNTs (Spetsmash, Kiev, Ukraine) have been synthesized by CVD method using Al2FeMo0,21 as a catalyst. The carbon content in the sample was 99.2% with soot as a residue; the catalyst was not found. The diameters of the MWCNTs varied from 2 to 40 nm; the surface area was 350 m2/g. The material has been certified by high-resolution transmission electron microscopy and Raman scattering [20].

A density of >650 mg cm-3 was used to define cortical bone Endos

A density of >650 mg cm-3 was used to define cortical bone. Endosteal and periosteal circumference were derived using a circular ring model. 4502 pQCT scans were performed, of which 88 were excluded due to major motion artifacts. Coefficients of variation

for pQCT scans, based on 139 subjects scanned a mean of 31 days apart, were 2.7%, 1.3% and 2.9% for BMCC, BMDC Roscovitine and cortical bone area, respectively. Other variables At 15.5 years research clinics, standing height (mm) was measured using the Harpenden Stadiometer (Holtain, Crymych, Wales, UK), and weight using the Tanita Body Fat Analyzer (model TBF 305; Tanita, Arlington Heights, IL, USA). Whole body DXA scans were performed using a Lunar Prodigy scanner with paediatric scanning software (GE Lunar Prodigy, Madison, WI, USA), providing measures of total body fat and lean mass. Maternal SEP was recorded at 32 weeks gestation by questionnaire and categorised according to the Office of Population Censuses and Surveys. Maternal GS-9973 research buy education was assessed at the same time by questionnaire. Pubertal stage was assessed using a Tanner stage (pubic hair domain) questionnaire completed

at age 14.7 years [22]. Moderate and vigorous physical activity was assessed by actigraph accelerometre at age 11, and subsequently found to be related to BMD in ALSPAC [23]. Date of birth and sex was obtained from birth notification, and date of the scan was recorded automatically, allowing age at scan to be calculated. Statistical analyses Descriptive statistics show means, standard deviation (SD), medians and lower and upper quartiles. Analyses were performed using seasonally adjusted 25 (OH)D3, which was modelled according to date of blood sampling using linear regression with trigonometric sine and cosine functions. 25(OH)D3 was loge transformed to reduce

heteroscedasticity. The residual was used as the primary 25(OH)D3 exposure variable in subsequent regression analyses. All analyses were performed C59 mw on standardised variables, i.e. subtracting the mean and dividing by the SD. To include all participants on whom a 25(OH)D2 was assayed, those with a value below the detectable limit of the assay (0.5 ng ml-1) were assigned a binary variable indicating whether an individual was at or below the lower limit, which was used as a covariable in all regression models. No individuals had 25(OH)D3 below the detectable limit of the assay. Models were checked for linearity by adding higher-order terms into the linear predictor and by comparing the likelihood of nested models. Further analyses were performed using a nonparametric bootstrap procedure in conjunction with OLS linear regression, based on 5,000 replications. Beta (β) estimates and standard errors were calculated from the mean and SD of the bootstrap distribution, respectively. All P values were calculated using bootstrap means and standard errors, compared to a Z-distribution and 95% percentile confidence intervals calculated.

Numerous gene target studies have shown the importance of CD4+ ac

Numerous gene target studies have shown the importance of CD4+ activation in resistance to Salmonella infection [41, 42]. Our data indicates a cellular immune response in mice immunized with the gidA mutant STM strain. Although the flow cytometric analysis showed no induction of memory T cells, or difference in CD8+ cells, it shows an increase in CD4+ population in the immunized mice at both day 7 and 42 post-immunization. It has been shown that CD4+ cells are more important than CD8+ in resistance to Salmonella infection [43,

44]. The passive transfer of cells to naïve mice from immunized mice did not confer full protection, and was not as significant as the serum passive transfer, but there was enough cell mediated immunity activated to protect a portion of the mice from a lethal dose challenge. Furthermore, Selleckchem ON-01910 splenocytes from immunized mice proliferated at a much higher rate than splenocytes from control mice when treated with STM cell lysate. The IgG1 induction was significantly more prominent than the induction of IgG2a, but the level of IgG2a was still significantly higher in the immunized mice than in that of the sera of the control mice. Furthermore,

the induction of the Th1 cytokines, IL-2 and IFN-γ, shows a strong indication of cell mediated immunity induced by immunization. In particular, IFN-γ showed a marked increase in cell culture find more supernatant when splenocytes from immunized mice were treated with STM cell lysate. The general consensus is that

the ideal Salmonella vaccine should generate both humoral and cell mediated immunity. This is due to protective immunity to Salmonella in mice being attributed to a balance between humoral and cell mediated immunity with an emphasis on development of the Th1 and Th2 subsets [45, 46]. In this study, the gidA mutant vaccine strain generated both Th1 and Th2 immunity with the Th2 immune response being the more prominent of the two. This was somewhat surprising since Salmonella is a facultative intracellular pathogen. One possible explanation for this could be found in our initial GidA study comparing the gidA mutant to the WT STM strain. The gidA mutant showed an approximate Anacetrapib 1000-fold reduction in the ability to invade T84 intestinal epithelial cells, as well as a marked reduction in ability to cause systemic infection in mice. Additionally, transcriptional and proteomic profiling identified a significant down-regulation in numerous genes and proteins responsible for invasion. Overall, the gidA mutant vaccine strain provides full protection to mice when challenged with a highly lethal dose of WT STM. The passive transfer experiments show the importance of both humoral and cell mediated immunity in this protective mechanism. This is an initial study in which a proof of principle of protective immunity has been established suggesting a gidA mutant STM strain could be a good candidate for use in a live-attenuated Salmonella vaccine.

Firstly, nucleotide sequences, as whole contigs were directly ali

Firstly, nucleotide sequences, as whole contigs were directly aligned using the MUMmer program [16]. Secondly, ORFs of a given pair of genomes were reciprocally compared each other, using the BLASTN, BLASTP and TBLASTX programs (ORF-dependent comparison). Thirdly, a bioinformatic pipeline was developed to identify

homologous regions of a given query ORF. Initially, a segment on a target contig homologous to a query ORF was identified using the BLASTN program. This potentially homologous region Selleck AZD2014 was expanded in both directions by 2,000 bp, after which, nucleotide sequences of the query ORF and selected target homologous region were aligned using a pairwise global alignment algorithm [40]. The resultant matched region in the subject contig was extracted and saved as a homolog (ORF-independent comparison). Orthologs and paralogs were differentiated by reciprocal comparison. In most cases, both ORF-dependent and -independent comparisons yielded the same orthologs, though the ORF-independent

method performed better for draft sequences of low quality, in which sequencing errors, albeit rare, hampered identification of correct ORFs. To determine average nucleotide (ANI) and average amino acid identities (AAI) for the purpose of assigning genetic distances between strains and strains to species groups, a recripocal best match BLASTN analysis was performed for each genome. The average similarity between genomes was measured Selleck Foretinib as the average nucleotide identity (ANI) and average amino acid identity (AAI) of all conserved protein-coding genes, following Fludarabine the methods of Konstantinidis and Tiedje [41]. By this method, AAI>95% and ANI>94% with >85% of protein-coding genes conserved between the pair of genomes, is judged to correspond to strains

of the same species, whereas AAI<95% and ANI <94% and <85% conservation of protein-coding genes indicate different species. Dinucleotide relative abundances were determined for each genome used in this analysis. Genomic dissimilarities between genomes were determined following the methods of Karlin et al. [42]. A multi-locus sequence analysis (MLSA) was determined following standard methods for the Vibrionaceae [21]. Data for the MLSA were reported as percent similarity between concatenated homologous ORFs for the genomes which encoded these ORFs. These criteria were applied to results of the analyses employed in this study. Identification and annotation of genomic islands Putative genomic islands (GIs) were defined as a continuous array of five or more ORFs discontinuously distributed among genomes of test strains following the methods of Chun et al [17]. Correct transfer or insertion of GIs was differentiated from deletion events by comparing genome-based phylogenetic trees and complete matrices of pairwise orthologous genes between test strains.

Adv Funct Mater 2013, 23:608–618 CrossRef 24 Scott TF, Kowalski

Adv Funct Mater 2013, 23:608–618.CrossRef 24. Scott TF, Kowalski BA, Sullivan AC, Bowman CN, McLeod RR: Two-color single-photon photoinitiation and photoinhibition for subdiffraction photolithography. Science 2009, 324:913–917.CrossRef 25. Li LJ, Gattass RR, Gershgoren E, Hwang H, Fourkas JT: Achieving λ/20 resolution by one-color initiation

and deactivation of polymerization. Science check details 2009, 324:910–913.CrossRef 26. Cao YY, Gan ZS, Jia BH, Evans RA, Gu M: High-photosensitive resin for super-resolution direct-laser-writing based on photoinhibited polymerization. Opt Express 2011, 19:19486–19494.CrossRef 27. Andrew TL, Tsai HY, Menon R: Confining light to deep subwavelength dimensions to enable optical nanopatterning. Science 2009, 324:917–921.CrossRef 28. Tanaka T, Sun HB, Kawata S: Rapid sub-diffraction-limit laser micro/nanoprocessing in a threshold material system. Appl Phys Lett 2002, 80:312–314.CrossRef 29. Thiel M, Fischer J, Freymann G, Wegener M: Direct laser writing of three-dimensional submicron structures using a continuous-wave laser at 532nm. Appl Phys Lett 2010, 97:221102.CrossRef 30. Qi FJ, Li Y, Tan DF, Yang H, Gong QH: Polymerized nanotips via two-photon photopolymerization. Opt. Soc. Am. 2007, 15:971–976. 31. Hell SW: Far-field optical

nanoscopy. Science 2007, 316:1153–1158.CrossRef 32. Kant R: Effect of primary spherical aberration on high-numerical aperture focusing of a Laguerre-Gaussian beam. J. Opt. Soc. Am. A 2008, MDV3100 25:1307–1318.CrossRef Competing interests The authors declare that they have no competing interests. Authors’ contributions CZ carried out specimen preparation, data acquisition and analysis of measurement and simulation and drafted the manuscript. KW, JB, GW and CG conceived the experiment, designed the plan and directed the drafting of the manuscript. SW, WZ and FY contributed to the simulation program improvement and participated in drafting the manuscript.

All authors read and approved the final manuscript.”
“Background Selleck Dolutegravir Industrial operations such as spin coating, painting, and lubrication are based on spreading of fluids over solid surfaces. The fluid may be simple [1–3] or particulate such as paint, ink, or dye [4]. For many years, capillary flow of simple fluids has received considerable attention, and physics of capillary action is known for a long time [5–9]. In addition, capillary flow of micellar surfactant solutions which contain monodisperse and naturally stabilized nanoparticles has been studied [10–14]. However, the same study on liquids laden with metallic and oxide nanoparticles such as silver, copper, zinc oxide, and titanium oxide is scarce. These fluid suspensions are termed as nanofluids after the seminal work by Choi and Eastman [15]. The application of nanofluids is coined with enhanced heat transfer performance compared with their base fluids.