It remains unclear whether FT actively suppresses innate immune responses during the early stages of infection, or if the delayed response is due to poor recognition of FT through host pattern recognition receptors. It has been

well documented that FT produces an atypical LPS that is not recognized via TLR4 [49–51] and that FT is recognized via the TLR2 signaling pathway [52–55]. Because the galU gene has been shown to be important for LPS production [27, 31, 32, 43, 56] in a selleck screening library number of other bacterial systems, we performed a series of studies to determine whether differences in the LPS expressed by the FT galU mutant might contribute to its reduced virulence. A western blot of both bacterial extracts and LPS preparations revealed no obvious differences in the O-antigen laddering between the galU mutant and WT strains of FT,

suggesting that mutation of galU did not have any gross effects on O-antigen synthesis. Because it has been reported elsewhere [57] and confirmed here (wbtA mutant) that the absence of O-antigen is a major determinant of susceptibility to complement-mediated killing, our findings that the galU mutant displayed a WT serum sensitivity phenotype also LY3023414 suggested that O-antigen synthesis was not significantly altered by mutation https://www.selleckchem.com/products/pnd-1186-vs-4718.html of the galU gene. This finding contrasted with reports that galU mutant strains of P. aeruginosa and V. cholerae displayed increased serum sensitivity [31, 44]. We also observed no differences between the galU mutant and WT strains of Teicoplanin FT with respect to signaling via the TLR2 and TLR4 recognition pathways. It remains possible that mutation of galU results in minor O-antigen compositional changes, alterations in the core oligosaccharides, or differences in the carbohydrate modification of surface proteins of FT. Moreover, in light of the published finding that mutations causing alterations in the lipid A of FT novicida [17, 20] are highly attenuating for virulence in vivo (possibly due

to altered kinetics of cytokine/chemokine production and neutrophil mobilization), we posit that mutation of the galU gene may have an impact on the lipid A moieties of FT. A complete analysis of the carbohydrate components of the FT galU mutant is needed to identify such differences. Recent studies have revealed that the innate immune response to FT infection is complex and involves multiple signaling pathways. Others and we have previously shown that FT elicits a powerful inflammatory response that is primarily mediated by TLR2 and caspase-1 activation [52–55]. More recently, it has been demonstrated that the AIM-2 inflammasome mediates caspase-1 activation and secretion of mature IL-1β and IL-18 during FT infection [42, 58, 59].

A protein strongly contributing to the stability of p53 is poly(ADP-ribosyl) polymerase-1 (PARP-1) [38, 42, 43], a protein that enzymatically modifies p53 [19, 41] thereby preventing its nuclear export [19, 39] by impeding the binding to CRM1 [19]. A protein that retains p53 in the cytoplasm preventing its nuclear functions, is mortalin, a member of the heat shock protein 70 (HSP70) family.

Mortalin binds p53 [31] and inhibits its pro-apoptotic functions what leads www.selleckchem.com/products/azd9291.html to increased tumor MLN2238 nmr development [31, 37]. The constitutive overexression of p53 in cells or animals is not feasible because this would trigger apoptosis or at least cell cycle arrest, making a functional study of the proteins’ features impossible. Fortunately, a temperature-sensitive (ts) mutant of p53 that displays wt properties at 32˚C but mutant character at elevated temperature [25], can be used to perform experiments aimed to elucidate its functions. This ts mutant demonstrates selleck screening library clear properties of mutant p53 at 39°C. At 37°C the cells also behave like mutant cells although a small portion of

p53 protein is in wt conformation. However, mutated p53 protein localized in the cytoplasm impedes the action of the wt protein. Thereby, the conformation and activity of p53 can be changed at will by simply growing the cells at 37 or 39˚C. The decision of p53 to trigger cell cycle arrest or apoptosis depends on the severity of the damage and is also regulated on the transactivational level by the use of p53 responsive elements to which the protein has different binding affinity [16]. In general, p53 binds to targets P-type ATPase mediating cell cycle arrest with a higher affinity

than to those which induce apoptosis [16]. A recent publication also showed that p53 is capable of inducing anti-apoptotic targets [17], adding further complexity to the functions and activities of the tumor suppressor protein. Also the Ras proteins are important for tumor development. In their active form they reside in the cytoplasmatic membrane and transmit signals from growth factor stimulation and downstream targets involve Raf-1 and PI3-kinase. Gain of function mutations lead to a constitutively active Ras protein that sustains growth-promoting signals, irrespective of extracellular stimulation, resulting in uncontrolled proliferation. For its proper anchoring in the cytoplasmic membrane and activity, Ras has to be isoprenylated by farnesyl protein transferases (FPTases) or/and geranylgeranyl protein transferases. Therefore, inhibitors of farnesylation have been used for treatment of cancers with constitutively activated RAS. Interestingly, tumor cells with constitutively activated RAS are rendered prone to treatment with pharmacological inhibitors of cyclin-dependent kinases (CDKs) like roscovitine (ROSC) and olomoucine (OLO) when they are pre-treated with FTIs [45].

In the present study, certain building-associated basidiomycetes including Serpula lacrymans (the causative agent of timber dry rot), Antrodia sitchensis, Trametes versicolor and Gloeophyllum sepiarium [45, 46], were found, mostly from the water-damaged, wood-framed Index-1 building. These species may have had an intramural source also in the present study. However, this connection could not be verified by examination of the building materials. Several opportunistically pathogenic taxa [47] were also identified, including Candida zeylanoides, Cryptococcus

see more albidus, Exophiala xenobiotica, Mucor spp. and Trichosporon mucoides. In addition to a wide diversity of fungi, we also found DNA signatures of an impressively diverse array of plants including cultivated crops (fruits, vegetable crops and tobacco), deciduous trees,

grasses, mosses and weeds. The amplification of plant DNA was likely due to a lack of specificity in our forward PCR primer [23]. Despite the fact that the inclusion of plant targets was not our intent, their recovery further confirms the biological complexity of dust, and indicates that DNA-based methods may be useful for the detection of dust-borne plant particles. Like fungal particles, those originating from I-BET151 price plants may also have allergenic potential, and obviously persist in indoor dust, long past the respective pollen season. The representativeness of different dust sample types has been discussed in the context of airborne exposure analysis; for example, Cediranib (AZD2171) the presence of heavy, non-resuspending particulate material in floor dusts, as well as potential microbial proliferation in dusts collected from locations with elevated relative humidity have been suspected to bias dustborne measurements [48–50]. A comparison of our above-floor surface samples with floor dust samples collected earlier during the cold season from the same

geographic region [23] indicated differences in fungal community composition. Especially, lower frequencies of basidiomycetous yeasts (mainly www.selleckchem.com/products/azd3965.html Malassezia and Cryptococcus) and rusts were found in dusts collected from elevated surfaces. This difference was also reflected in the differential ratios of Ascomycetes and Basidiomycetes (NAsc:NBas) between the two sample types; the average NAsc:NBas ratio was 3.03 for the elevated surface dust, but lower (0.95) for floor dust. The differences may relate to the aerodynamic properties of different fungal particles; while the spores of the mentioned genera are not distinguishingly large, they are commonly carried along with larger particles (i.e. Malassezia cells on human skin scales and Cryptococcus cells on plant debris), which makes them more prone to deposit on floor surfaces. In contrast, many ascomycetous particles are small, air-dispersed microconidia that stay airborne for long periods, resuspend efficiently and deposit on elevated surfaces.

10 ml samples were centrifuged, washed with 10 ml RN media and 10 ml H2O. Pellets were resuspended in 100 μl H2O and lipids extracted through the addition of 360 μl of chloroform:methanol:HCl (1/2/0.02) and incubated at room temperature for 20 minutes. 120 μl chloroform and 120 μl 2 M KCl were added to separate

phases and after centrifugation, the organic phase was removed and radioactivity quantified by scintillation counting. Thin-layer chromatography Radiolabelled lipids were analyzed by 1-dimensional and 2-dimensional thin-layer chromatography. The 1-dimensional system used to separate phospholipids SBI-0206965 from diacylglycerol and fatty acid was Silica Gel G layers developed with chloroform:methanol:acetic acid (98/2/1) and visualized using Bioscan imaging detector. The 2-dimensional

system also employed Silica Gel G layers and was developed first with chloroform:methanol:water (65/25/4) and secondly tetrahydrofuran/dimethoxyethane/methanol/4 M ammonium hydroxide (10/6/4/1). The resulting thin-layer plate exposed Belnacasan ic50 to a PhosphoImager screen and visualized using a Typhoon 9200. Lipid mass spectrometry Mass spectrometry of phospholipids was performed using a Finnigan TSQ Quantum (Thermo Electron, San Jose, CA) triple quadrupole mass spectrometer. Samples were prepared in 50:50 (v/v) chloroform:methanol. The instrument was operated in the negative ion mode. Ion source oxyclozanide parameters were as follows: spray voltage

of 3,000 V, capillary temperature of 270°C, capillary offset of 35 V, and the tube lens offset was set by infusion of polytyrosine tuning and calibration solution (Thermo Electron, San Jose, CA) in the electrospray mode. Acquisition parameters were as follows: full scan, scan range 600 – 100 m/z, scan time 0.5 s, peak width Q1 0.7 FWHM. Instrument control and data acquisition was performed with the Finnigan™ Xcalibur™ software (Thermo Electron, San Jose, CA). Mass spectrometry malonyl-CoA measurement Cultures of strain PDJ28 were grown in RN medium supplemented with 0.1% glycerol to OD600 = 0.6. Cells were pelleted and washed with 50 ml RN medium to remove glycerol and used to inoculate RN medium with and without 0.1% glycerol. Cultures were grown for 120 minutes and harvested at room temperature. Cells were extracted using the Bligh and Dyer method [27], and 50 pmol of [13C3]malonyl-CoA (Stable Isotope Products; Isotec) was added. The aqueous phase was applied to a 100-mg 2-(2-pyridyl)ethyl functionalized silica gel column (Supelco) equilibrated with 2% acetic acid in methanol/water (1:1) [28]. The column was washed two times with 1 ml of equilibration buffer and 1 ml water. CoAs were eluted with 1 ml of 50% acetonitrile containing 15 mM ammonium hydroxide. Mass spectrometry of acyl-CoA was performed using a Finnigan TSQ Quantum (Thermo Electron) 10058-F4 triple-quadrupole mass spectrometer [29].

After 48-72 h the parasites were harvested in PBS and centrifuged (200 g for 7-10 min) at room temperature in order to

discard blood cells and cellular debris. The supernatant was collected and then centrifuged again at 1000 g for 10 min. The final pellet was resuspended in DMEM and used in the interaction assays. T. gondii infection Proteases inhibitor during skeletal muscle cell myogenesis Aiming to verify the infectivity of T. gondii in myoblasts and myotubes, we developed the following protocol: 2-day-old cultures were infected with tachyzoite forms (1:1 parasite-host cell ratio) and, after 24 h of interaction, the total number of infected myoblasts and myotubes was quantified independent of the number of internalized parasites. For evaluation of the potential interference of T. gondii in myotube formation, after the initial seeding, cultures were maintained for 48 h in medium without calcium, in order to not stimulate myoblast fusion. Ganetespib manufacturer After this time, the cultures, enriched in myoblasts, were infected for 24 h. Cell fusion in the presence or absence of T. gondii was determined by morphological analysis of myoblast alignment and the observation of the percentage of multinucleated cells. The quantitative analysis was based on 3 independent experiments performed in duplicate with at least

200 cells in each phosphatase inhibitor coverslip. Fluorescence analysis of actin microfilaments SkMC 2-day-old cultures were allowed to interact with tachyzoites (1:1 parasite: host cell ratio) for 24 and 48 h at 37°C. Non-infected and infected SkMC were fixed for 5 min at room temperature in 4%

paraformaldehyde (PFA) diluted in PBS. After fixation, the cultures were washed 3 times (10 min each) in the same buffer. Then, the cultures were incubated for 1 h at 37°C with 4 μg/ml phalloidin-rhodamine diluted in PBS. Thereafter, the cultures were washed 3 times (10 min each) in PBS, incubated for 5 min in 0.1 μg/mL DAPI (4′,6-diamidino-2-phenylindole, Sigma Chemical Co.), a DNA stain that enables the visualization of host and parasite nuclei, and washed again in PBS. The coverslips were mounted on slides with a Lepirudin solution of 2.5% DABCO (1,4-diazabicyclo-[2]-octane-triethylenediamine antifading, Sigma Chemical Co.) in PBS containing 50% glycerol, pH 7.2. The samples were examined in a confocal laser scanning microscope (CLSM Axiovert 510, META, Zeiss, Germany) from the Confocal Microscopy Plataform/PDTIS/Fiocruz, using a 543 helium laser (LP560 filter) and 405 Diiod laser (LP 420 filter). Immunofluorescence analysis of total cadherin protein distribution in SkMC myogenesis during infection with T. gondii Immunofluorescence assays were performed using specific monoclonal antibodies for pan-cadherin (Sigma Chemical Co. C3678). Briefly, tachyzoite forms were allowed to interact with 2-day-old SkMC in the ratio of 1:1.

During FIRST, the calcium and vitamin D status of all women was assessed, and they were given daily supplements of up to 1,000 mg of selleck chemicals elemental calcium and up to 800 IU of vitamin D for a period of 2 weeks to 6 months. Supplementation doses and duration were adjusted for each patient according to their baseline calcium and 25-OH vitamin D status. After the run-in period, eligible women were proposed for enrolment in either the SOTI or TROPOS studies,

and supplementation was continued at the same doses throughout the randomised treatment periods of both these studies. The SOTI study included women ≥50 years of age with low lumbar BMD (<0.840 g/cm2 measured with Hologic instruments, T-score ≤−2.4) and at least one prevalent PRI-724 manufacturer vertebral fracture confirmed by spinal radiography. The TROPOS study included women with femoral

neck BMD <0.600 g/cm2 and aged ≥74 years or 70–74 years with one additional risk factor (history of osteoporotic fracture after menopause, residence in a retirement home, frequent falls or maternal history of osteoporotic fracture of the hip, spine or wrist). Study design and efficacy measurements Patients were randomised to receive strontium ranelate 2 g/day or placebo for 5 years (TROPOS) selleck chemical or 4 years followed by a 1-year treatment-switch period (SOTI). In both studies, main efficacy analyses were performed at 3 years, and the vertebral fracture data over 3 years were used for the present analysis. Baseline refers to the commencement of the SOTI and TROPOS studies, SPTBN5 not the time of inclusion in FIRST. Vertebral fractures were determined from radiographs taken at baseline and annually thereafter and were analysed in the same way in both studies. Radiographs were analysed by the semi-quantitative method of Genant et al. [22, 23], using a four-point grading scale: grade 0—normal; grade 1—mild deformity (20–25% decrease in at least one vertebral height); grade 2—moderate deformity (25–40% decrease); and grade 3—severe deformity (>40% decrease). A new vertebral fracture was defined as a change

from a non-fractured vertebra (grade 0) to a vertebra rated grade 1 or higher. All radiographs were analysed at a central facility (CEMO, France) blinded to treatment assignment but not to temporal sequence. Lumbar L2–4 and femoral neck BMD were measured at baseline, and lumbar BMD was measured every 6 months post-baseline by dual-energy X-ray absorptiometry using Hologic devices. All scans were analysed centrally, and a programme of cross-calibration across centres was performed throughout both studies [24]. Blood samples were collected at baseline, 3 months, 6 months, and then every 6 months. Serum samples were stored at −80°C and analysed centrally after a maximum 6 months period of storage (University of Liège, Belgium).

v-viCrossRef 8. Billat VL, Demarle A, Slawinski J, Paiva M, Koralsztein JP: Physical and training characteristics of top-class marathon runners. Med Sci Sports Exerc 2001,33(12):2089–2097.check details PubMedCrossRef 9. di Prampero PE, Atchou G, Bruckner JC, Moia

C: The energetics of endurance running. Eur J Appl Physiol Occup Physiol 1986,55(3):259–266.PubMedCrossRef 10. Rapoport BI: Metabolic factors limiting performance in marathon runners. PLoS Comput Biol 2010,6(10):1–13.CrossRef 11. Hargreaves M, Angus D, Howlett K, Conus NM, Febbraio M: Effect of heat stress on glucose kinetics during exercise. J Appl Physiol 1996,81(4):1594–1597.PubMed 12. Sawka MN, Burke LM, Eichner ER, Maughan RJ, Montain SJ, Stachenfeld NS, American College of Sports M: American College of Sports Medicine position stand. Exercise and fluid replacement. learn more Med Sci Sports Exerc 2007,39(2):377–390.PubMedCrossRef 13. Bar-Or O: Effects of age and gender on sweating pattern during exercise. Int J Sports Med 1998,19(Suppl 2):S106-S107.PubMedCrossRef

14. Kelley GA, Lowing L, Kelley K: Gender differences in the aerobic fitness levels of young African-American adults. J Natl Med Assoc 1999,91(7):384–388.PubMedCentralPubMed 15. Mehnert P, Brode P, Griefahn B: Gender-related difference in sweat loss and its impact on exposure limits to heat stress. Int J Ind Ergonom 2002,29(6):343–351.CrossRef 16. Kaciuba-Uscilko H, Grucza R: Gender differences in thermoregulation. Curr Opin Clin Nutr Metab Care 2001,4(6):533–536.PubMedCrossRef 17. Hessemer V, Bruck K: Influence of menstrual-cycle on shivering, skin blood-flow, and sweating responses measured at night. J Appl Selleck Nutlin 3 Physiol 1985,59(6):1902–1910.PubMed Caspase inhibitor 18. Toner MM, Sawka MN, Foley ME, Pandolf KB: Effects of body mass and morphology on thermal responses in water. J Appl Physiol 1986,60(2):521–525.PubMedCrossRef 19. Gagge AP, Stolwijk JAJ, Hardy JD: Comfort and thermal sensations and associated

physiological responses at various ambient temperatures. Environ Res 1967,1(1):1–20.PubMedCrossRef 20. Glickman EL, Peacock C, Gunstad J, Kakos L, Burns KJ, Pollock B, Feeback M, Seo Y: A thermal perception scale for use during rest and exercise in 37°C ambient air [abstract]. Med Sci Sports Exerc 2013,45(5):S70. 21. Armstrong LE: Exertional Heat Illnesses. Champaign, IL: Human Kinetics; 2003. 22. Brooks GA, Fahey TD, Baldwin KM: Exercise Physiology: Human Bioenergetics and Its Applications with PowerWeb Bind-in Card. New York, NY: McGraw-Hill Higher Education; 2004. 23. Davis JM, Burgess WA, Slentz CA, Bartoli WP, Pate RR: Effects of ingesting 6% and 12% glucose/electrolyte beverages during prolonged intermittent cycling in the heat. Eur J Appl Physiol Occup Physiol 1988,57(5):563–569.PubMedCrossRef 24. Armstrong LE: Assessing hydration status: the elusive gold standard. J Am Coll Nutr 2007,26(Suppl 5):575S-584S.PubMedCrossRef Competing interests The authors declare that they have no competing interests.

microplus was eradicated like the USA. Conclusion Tick microbiomes remain largely unexplored. By EPZ-6438 cell line comparison to the proposed strategy to

accomplish the Human Microbiome Project, the work presented here constitutes the initial data acquisition and analysis exercise towards a comprehensive analysis of the R. microplus microbiome. A thorough understanding of the functional, ecological, and evolutionary aspects of the bacterial diversity in communities associated with the cattle tick requires additional investigations. The bacteria we found could have favorable effects on the tick’s successful infestation of its cattle host, perhaps with roles in host blood CB-839 manufacturer digestion, immunity against infection by competing microbes potentially deleterious to the tick, or

effects on population structure and fertility. Cattle AR-13324 ticks have evolved in conjunction with bovine hosts; therefore, bovine-tick interactions have likely influenced the ecology of their microbiomes. Even within the tick itself, there are feedback mechanisms influencing interactions at the host-microbiome interface. Our results further document the co-infection of cattle ticks with several bacteria, even in the presence of antimicrobial factors that are known to be produced by the tick immune system response in their hemolymph and gut tissues. Further investigations on the cattle tick microbiome are likely to enhance our understanding of the roles this cosmopolitan species serves as vector of bacteria that

may be pathogenic to its vertebrate hosts. Methods Tick samples Adult male and female ticks were obtained from a R. ifenprodil microplus infestation outbreak on cattle from Starr County, TX. Samples from the infestation were collected by USDA personnel in November, 2008, and shipped to the USDA Cattle Fever Tick Research Laboratory in Moore Field, TX, where the samples were frozen at -80°C. Prior to freezing, eggs were collected from gravid females, mixed together, and pooled and labeled as f1 generation. A portion of these f1 eggs were used to establish a laboratory colony to obtain adult ticks as described previously [79]. Two adult females and two adult males developed from these f1 eggs and three small clumps of approximately 100 f1 eggs each were used for the DNA extraction and pyrosequencing. The gut and ovary samples were obtained from the f20 generation of the La Minita strain of R. microplus that has been maintained Babesia -free at the University of Idaho Holm Research Center since 1999. The founding ticks for this strain were originally collected in Starr County, TX, in 1996. Using standard protocols approved by the University of Idaho Institutional Animal Care and Use Committee, La Minita larvae were placed on a stanchioned calf and replete females collected and dissected under sterile phosphate-buffered saline during the period of active oviposition.

Authors’ contributions SHC carried out the preparation of AuNPs, AgMSs, AgMSs@GNPs assembly, Raman and XRD, characterization, drafted the manuscript, PH modified the draft of manuscript, ZHW carried out the UV and SEM Characterization. ZW checked the manuscript grammar. MS participated

click here in the analysis of Raman results. GN gave many advices for this manuscript. DXC and XYC designed of the study and guided this work. All authors read and approved the final manuscript.”
“Background Atomic layer deposition (ALD) facilitates the deposition of a dielectric oxide onto a GaAs surface. The process differs from the one used for the deposition of ALD oxide on Si, where an OH group on the semiconductor is required to initiate the deposition. Bonding of the oxide on the III-V semiconductor is accessible to investigation with high-resolution synchrotron radiation photoemission. It provides unprecedented, precise information about the interfacial electronic structure. This information is vital because the interfacial trap density (D it) governs the performance of selleck chemicals GaAs-based devices. In order to obtain consistent information, the III-V surface must be free of impurities, such as oxygen, and other defects prior to the ALD process. Only when this condition is satisfied will the true interfacial electronic structure be revealed. The attempt to DZNeP chemical structure prepare a

clean GaAs(001) surface has generally been patterned on the procedure used to obtain a clean Si(001) surface. That neglects the fundamental difference Sclareol between the surface properties and reconstruction of a III-V semiconductor and an elemental one like Si. The reconstructed Si(001)-2 × 1 surface consists of rows of buckled dimers, with charge transfer between the tilted

atoms, and is rich in dangling bonds that trap impurities. Surface pretreatment is required prior to a final anneal in an ultra-high-vacuum end station prior to synchrotron radiation photoemission (SRPES) measurements. The pretreatment due to Ishizaka and Shiraki [1] has come into general use. It leaves a thin oxide film on a clean Si surface that is readily removed by annealing in vacuum [2, 3]. The effectiveness of this procedure has been demonstrated in [2], which shows the analysis of 2p core-level data from a clean reconstructed Si(001) surface. The photoemission spectra from the first three surface layers labeled S(0), S(1), and S(2) are identified and have intensities consistent with the expected escape depth. For multi-element (In)GaAs, a common method of surface pretreatment prior to in-vacuum annealing is As capping [4] by thermal annealing in As2 flux [5], followed by a chemical rinse [6]. Subsequent in-vacuum annealing of these samples removes the more volatile As and produces an oxygen-free surface, but one that does not have the desired surface Ga/As ratio. It turns out to be low, say 0.73 [4], compared with an untreated sample, say 1.26 (not shown).

All annealing treatments were carried out in air in a box furnace with the substrates contained in a high-purity alumina crucible. In this study, the surface morphology was examined using an atomic force microscope (AFM; Veeco DID3100, Plainview, NY, USA) and scanning electron microscope (SEM; Hitachi S-4700, Tokyo, Japan). Results and discussion Top-view SEM micrograph of

soft mold (PDMS diluted with toluene) PU-H71 ic50 molding from the quartz master is shown in Figure 3a. As shown in Figure 3a, the patterned PDMS with 550-nm-wide lines separated by 250-nm space were obtained on the surface. The result of the UV curing imprinted pattern used by the replicated soft PDMS mold on the quartz master is shown in Figure 3b. It is easily seen that the patterned AMONIL-MMS4 VX-680 order TSA HDAC nmr with 250-nm-wide and 120-nm-long lines separated by 550-nm space was obtained on the Al thin film surface, which is coincident with that of the quartz master. The residual polymer layer with 60-nm thickness was removed by RIE. The patterns were subsequently transferred into Al thin films by RIE. Top-view SEM micrograph of patterned Al thin films obtained by the UV-NIL and RIE is shown in

Figure 3c. As shown in Figure 3c, the patterned Al thin films with 250-nm-wide lines separated by 550-nm space were obtained on the sapphire surface, which is coincident with that of the quartz master. Figure 3 SEM images of the morphology of PDMS soft mold molding. From the quartz master (a), patterned AMONIL-MMS4 (b), and patterned Al thin ADP ribosylation factor films obtained by the UV-NIL and RIE (c). Dramatic changes in the pattern morphology were observed following high-temperature annealing applied to induce grain growth of the sapphire. Figure 4a shows a SEM image of the morphology of the patterned surface after annealing for 24 h at 450°C and 1 h at 1,200°C. For nanopatterned Al thin

films that subsequently experienced an annealing temperature of 1,200°C, it was found that smoothing and coalescence of the line features had occurred to such an extent that the patterning was no longer discernible. The phenomenon of surface diffusion-driven smoothing of surface features is well established in the literature [19–22] and occurs due to surface energy considerations [23, 24]. The kinetics of the smoothing of the line patterns can be used to derive information on the diffusion mechanism. Therefore, for the successful fabrication of NPSS, the relative kinetics of smoothing versus grain growth of the underlying sapphire is critical. Fortunately, for high-temperature annealing at 1,000°C and 1,100°C, the patterns were retained on sapphire substrates. Figure 4b shows a SEM image of the morphology of the patterned surface after high-temperature annealing for 1 h at 1,000°C. Figure 4c shows the AFM image of nanopatterned Al thin films with 250-nm-wide lines separated by 550-nm space after dual-stage annealing for 24 h at 450°C and 1 h at 1,000°C.