Adv Mater 2009, 21:4087–4108 CrossRef 11 Zhang Q, Cao G: Nanostr

Adv Mater 2009, 21:4087–4108.CrossRef 11. Zhang Q, Cao G: Nanostructured photoelectrodes for dye-sensitized buy Semaxanib solar cells. Nano Today 2011, 6:91–109.CrossRef 12. Martinson ABF, Elam JW, Hupp JT, Pellin MJ: ZnO nanotube based dye-sensitized solar cells. Nano Lett 2007, 7:2183–2187.CrossRef 13. Zhang Q, Myers D, Lan J, Jenekhe SA: Applications of light scattering in dye-sensitized solar cells. Phys Chem Chem Phys 2012, 14:14982–14998.CrossRef 14. Wang ZS, Kawauchi H, Kashima T, Arakawa H: Significant influence of TiO2 photoelectrode morphology on the energy conversion efficiency

of N719 dye-sensitized solar cell. Coord Chem Rev 2004, 248:1381–1389.CrossRef 15. Kang SH, Kim JY, Kim HS, Koh HD, Lee JS, Sung YE: Influence of light scattering particles in the TiO2 photoelectrode for solid-state dye-sensitized solar cell. J Photochem Photobiol A 2008, 200:294–300.CrossRef 16. Ito S, Nazeeruddin M, Liska P, Comte P, Charvet R, Péchy P, Jirousek M, Kay A, Zakeeruddin S, Grätzel M: Photovoltaic characterization of dye-sensitized solar cells: effect of device masking on conversion efficiency. Prog Photovolt Res Appl 2006, 14:589–601.CrossRef 17. Hore S, Vetter C, Kern R, Smit H, Hinsch A: Influence of scattering layers on efficiency of dye-sensitized solar cells. Sol Energy Mater Sol Cells 2006, 90:1176–1188.CrossRef 18. Ito S, Nazeeruddin M, Zakeeruddin S, Péchy P, Comte P, Grätzel M, Mizuno T, Tanaka A, Koyanagi T: Study

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20. Qiu Y, Chen W, Yang S: Double-layered photoanodes from variable-size anatase TiO2 nanospindles: a candidate for high-efficiency dye-sensitized solar cells. Angew Chem Int Ed 2010, 49:3675–3679.CrossRef 21. Tan B, Wu YY: Dye-sensitized solar cells based on anatase TiO2 nanoparticle/nanowire composites. J Phys Chem B 2006, 110:15932–15938.CrossRef 22. Kevin M, Fou YH, Wong ASW, Ho GW: A novel maskless approach towards aligned, density modulated and multi-junction ZnO nanowires for enhanced surface area and light trapping solar cells. Nanotechnology 2010, 21:315602–315610.CrossRef 23. Tetreault N, Horvath E, Moehl T, Brillet J, Smajda R, Bungener S, Cai N, Wang P, Zakeeruddin SM, Forro L, Magrez A, Grätzel M: High-efficiency solid-state dye-sensitized solar cells: fast charge extraction through self-assembled 3D fibrous network of crystalline TiO2 nanowires. ACS Nano 2010, 4:7644–7650.CrossRef 24. Lin CJ, Yu WY, Chien SH: Effect of anodic TiO2 powder as additive on electron transport properties in nanocrystalline TiO2 dye-sensitized solar cells. Appl Phys Lett 2007, 91:233120.CrossRef 25.

aeruginosa mutants, KG7004 and KG7050, lacking quorum sensing and

aeruginosa mutants, KG7004 and KG7050, lacking quorum sensing and efflux protein genes were constructed by allele exchange using the plasmids listed in Table 1, as described previously [30, 35, 42]. Construction of P. aeruginosa mutants in this study followed the order: PAO1 to KG7001 with plasI (for deletion of lasI), KG7001 to KG7004 with pAF2071 (for deletion of rhlI), and KG7004 to KG7050 with pMexB (for deletion of mexB), respectively. Construction of QS reporter strains pSQG was

selleck screening library constructed by subcloning a 700-bp EcoRI digested fragment derived from pGreen into the KpnI site of mini-CTX1 [38, 39]. A lasB promoter-gfp translational fusion was constructed by ligating a 591-bp fragment including the region encoding N-terminal ten amino acids of LasB derived from the P. aeruginosa PAO1 chromosome. The resulting plasmid, pSQG003, was mobilized into KG7004 and KG7050 via E. coli S17-1. To accomplish excision, pFLP2, encoding Flp recombinase, was introduced into the P. aeruginosa KG7403 and KG7503

strains containing the lasB promoter-gfp translational fusion constructs by using the high transformation method and previously described procedures [40, 43]. In addition, the multicopy reporter plasmid pMQG003 was constructed. A lasB promoter-gfp translational fusion fragment from pSQG003 was cloned into pME6012 [41]. The lasB promoter-gfp translational fusion fragment Pritelivir solubility dmso was prepared by using PCR with the primers CTX1-F (5′-CGATAGATCTGCCGTCCTTGCTGAATTAGC-3′) and CTX1-R (5′-AACTAGATCTCGCTTTTGAAGCTGATGTGC-3′) containing an engineered restriction site BglII (forward and reverse). This fragment was restricted with BglII, and then ligated to the

BglII site of pME6012. Construction of the plasmids expressing the wild-type and mutant mexB genes in P. aeruginosa The stable E. coli–P. aeruginosa shuttle vector Rebamipide pKTA113 carrying mexB was constructed in three steps. The first mexB fragment amplified by PCR using the chromosomal DNA of P. aeruginosa PAO1 as a template and a pair of primers containing the engineered restriction sites HindIII (5′-ACATAAGCTTATGTCGAAGTTTTTCATTGATAGG -3′) and SalI (5′- GCAATCTAGATTGCCCCTTTTCGACGGACG -3′). Next, mexB fragments were ligated to the multicloning site of pUC18 to yield pYT06. To obtain the MexB expression plasmid, a 3138-bp HindIII-XbaI fragment from pYT06 was ligated to the large HindIII-XbaI fragment of pTO003. The resulting construct containing MexB-6His under the lac promoter shall be referred to as pKTA113 in this paper. To produce mexB mutants, the Stratagene Quickchange site-directed mutagenesis kit (Stratagene) was used according to the manufacturer’s protocol. The Phe136Ala or Asp681Ala substitution was introduced into pYT06, respectively. Then the mutated mexB fragments of the pYT06 mutants were subcloned into pTO003.

Cells were harvested by centrifugation and resuspended in SDS sam

Cells were harvested by centrifugation and resuspended in SDS sample buffer (SSB) [21] according to the following formula: resuspension volume (in μl) = 100 × A600 × vol harvested (in LY3009104 solubility dmso ml). These concentrated cell lysates were diluted 1:100 in SSB for SDS-PAGE. Cell-free supernatants were concentrated ~10-fold by filtration using Centricon spin columns (Millipore, Billerica, MA, USA), and added to concentrated SSB for SDS-PAGE. Samples were

separated on 4-12% SDS-polyacrylamide gels and stained with selleck chemicals llc silver to visualize protein bands [21]. SslE secretion experiments were repeated 2–4 times, and single representative gels are shown. To produce the images in Figure 2, the stained gels were digitally photographed

and gel images were enhanced using Adobe Photoshop software. Linear transformations (contrast and brightness adjustments) were applied to the images for clarity; such transformations were applied uniformly across any given gel image. Fusion protein localization by enzyme activity To measure secretion and surface display of SslE-enzyme fusions, cultures of WT and ΔpppA::FRT strains bearing the indicated plasmids were grown in LB at 37°C with aeration for 16–20 hours. Cells were harvested by centrifugation, and cell-free supernatants were removed; an aliquot of collected cells was removed and lysed using the PopCulture reagent from Novagen (Madison, WI, USA). Enzymatic activities associated with intact cells, lysed cells, and cell-free supernatants were then immediately measured. SslE-Cel45A SCH727965 chemical structure activity was measured using the CRACC assay [27], and Sitaxentan SslE-Pel10A activity was measured using the pectate lyase assay described by Collmer [28]. Growth comparisons Phenotypic microarray experiments were performed

using an OmniLog reader (Biolog, Hayward, CA, USA) as per the manufacturer’s instructions using plate types PM-9 and PM-10. Cultures were grown at 37°C for 48 hours, and respiration data were analyzed using the PM software provided with the OmniLog reader. Strains used were wild-type W and Δgsp::FRT (unmarked deletion of gspC-M). To compare urea tolerances in 96-well plates, wild-type, Δgsp::FRT, and ΔpppA::FRT strains were cultured in 200 μl aliquots of LB containing 0, 0.9 M, or 1.15 M urea in 96-well plates (inoculated as 1:100 dilutions from LB overnight cultures). Plates were grown with shaking at 37°C in a Tecan M1000 plate reader (Durham, NC, USA). Growth and survival were followed by regular measurement of A595 for each culture. To compare urea tolerances in glass culture tubes, wild-type, Δgsp::FRT, and ΔsslE::FRT strains were cultured in 8 ml volumes of LB containing no urea or 1.15 M urea on a rolling wheel at 37°C. Biological duplicate cultures of each strain were inoculated with 1:1000 dilutions from LB overnight cultures after verification that all overnight cultures grew to equivalent A600 turbidity readings.

5–4 5 Gy (dose) Figure 2M–P presents the minimum growth rate (27

5–4.5 Gy (dose). Figure 2M–P presents the minimum growth rate (27–58%) for 42 h obtained using the irradiation parameters, 90 MeV u-1 (energy), 60–120 keV μm-1 (LET), and 1.5–4.5 Gy (dose). These data suggest that the cellular growth rate of the D. natronolimnaea svgcc1.2736 strain is dependent on the irradiation energy of the 12C6+ions. Significant differences in the effects of 12C6+ ions at the same doses were also observed. This suggests a strong dependence of low-dose effects on LET (Figure 2I-L). Figure 2 12 C 6+ -ions see more of different parameters irradiation level and

its effect on the growth rate of D. natronolimnaea smgcc1.2736 strains cells in %. (A-D) 12C6+-ions were accelerated up to 30 MeV/u, and their LETs were 60, 80, 100 and 120 keV/μm, with a dose rate of 0.5-1.5Gy. (E-H) 12C6+-ions were accelerated up to 45 MeV/u, and their LETs were 60, 80, 100 and 120 keV/μm, with a dose rate of 0.5-1.5Gy. (I-L) 12C6+-ions were accelerated up to 60 MeV/u, and their LETs were 60, 80, 100 and 120 keV/μm, with a dose rate of 0.5-1.5 Gy. (M-P) 12C6+-ions were accelerated up to

90 MeV/u, and their LETs were 60, 80, 100 and 120 keV/μm, with a dose rate of 0.5-1.5 Gy. Effect of irradiation dose on productivity of D. natronolimnaea svgcc1.2736 Different irradiation doses showed a notable affect on the growth rate and conidia aggregation in D. natronolimnaea svgcc1.2736. CX production in 1 L cultures of D. natronolimnaea svgcc1.2736 mutants was, shown to be sensitive to irradiation dose

(Figure 3). XAV-939 manufacturer Overall, for CX producing strains of D. natronolimnaea svgcc1.2736 mutants, increasing the irradiation dose from the standard 0.5 to 4.5 Sepantronium concentration Gy led to a considerable much decline in dry cell weight (BDW), from around 8.71 ±0.04 to 2.23 ±0.06 g L-1, respectively. The CX yield, however, showed an almost two-fold increase from 8 ±0.9 to 12 ±0.2 mg L-1. To find the optimal 12C6+ irradiation dose for the process, a considerable amount of cell culture was carried out using similar irradiation experiments. natronolimnaea svgcc1.2736 strains productivity increases by almost six-fold. Optimal production of 0.81 mg L-1 h-1 was detected at a irradiation dose of approximately 4.5 Gy at an 80 keV μm-1 LET and 60 MeV u-1 energy level (Figure 3B). In contrast, 12C6+ irradiation with a LET of more than 100 keV μm-1, and energy level of greater than 45 MeV u-1 reduced the rate of production (Figure 3D). 12C6+ irradiation with LET (80 keV μm-1), energy (60 MeV u-1) and dose (1.5 Gy) led to perfect mycelial growth (Figure 3A). The increased irradiation dose of 12C6+ however led to a decrease in biomass in this strain (Figure 3). Figure 3B depicts the BDW and productivity of the strains with respect to different energy (45 and 60 MeV u-1) versus an irradiation dose with a LET of 80 keV μm-1.

Supplements that were defined as “”herbal supplements”" were prod

selleckchem supplements that were defined as “”herbal supplements”" were products mainly derived from plant sources such as echinacea, garlic and ginseng. “”Other supplements”" included products that couldn’t be categorized any other way, such as

fibres, beastings and conjugated linoleic acid. “”Vitamin supplements”" included multivitamins, vitamins A, B, C, D and E, beta-carotenes and antioxidant agents. “”Mineral supplements”" consisted of iron, calcium, magnesium and other mineral products such as zinc, fluorine, potassium and multi-minerals. Statistical methods Odds ratios (ORs) for use of dietary supplements and their 95% CIs AG-881 for athlete subgroups in 2009, compared with athlete subgroups in 2002, were analyzed using logistic

regression model with the aid of SPSS 16.0 software. Age, sex and type of sport were included in the analysis as independent covariates. Results Frequency of supplement use in 2002 and 2009 The questionnaire p53 activator was completed by 446 of 494 (90.3%) athletes in 2002 and 372 of 405 (91.7%) athletes in the follow-up study. Of the 446 athletes, 81% reported supplement use during previous 12 months in 2002 and 73% of the 372 athletes in 2009. Decreased consumption of dietary supplements between study years was seen in all subgroups except for amino acids (3.8% in 2002 and 7.3% in 2009), oils and fatty acids (11% and 19%), homeopathic supplements (0.4% and 1.6%), multivitamins (54% and 57%) and antioxidants (0.7% and 2%). Differences in supplement use Selleck Baf-A1 between study years are illustrated in Figure 1. Dietary supplement use in different sports in 2002 and 2009 are illustrated in Figures 2 and 3. Figure 1 Dietary supplement use between study years. Figure 2 Dietary supplement

use in different sports in 2002. Figure 3 Dietary supplement use in different sports in 2009. Mean number of supplements consumed were 3.4 ± 3.1 in 2002 and 2.6 ± 2.7 in 2009. In 2002, the highest amount of different dietary supplements consumed per athlete was 18. In 2009, the highest amount of different dietary supplements was 14. In 2009, among all athletes the most often declared subgroup used was vitamin supplements (56%) and most of the vitamin supplement users consumed multivitamins (57%). Nutritional supplements were used by 52% of the athletes, proteins (38%) and oils and fatty acids (19%) being the biggest subgroups. All dietary supplement use After adjusting for age-, sex- and sport type, the OR (95% CI) for use of any dietary supplement was significantly less in 2009 sample as compared with 2002 sample (OR, 0.62; 95% CI, 0.43-0.90). Athletes in speed and power events and endurance events reported use of any dietary supplement significantly more often than team sport athletes both in 2002 and 2009 (Table 3). In 2002, all DS use among athletes in skill-based sports was significantly less than among athletes in team sports (OR, 0.46; CI 0.25-0.85).

Study subject

Study subject Selleck MCC-950 The subjects of this study included all patients who

were operated for perforated peptic ulcers at Bugando Medical Centre during the period under study. Patients with incomplete data were excluded from the study. Patients treated conservatively and those who failed to consent for HIV infection were also excluded from the study. The details of patients who presented from April 2006 to March 2008 were retrieved retrospectively from patient registers kept in the Medical record departments, the surgical wards, and operating theatre. Patients who presented to the A & E department between April 2008 and March 2011 were prospectively enrolled in the study after signing an informed written consent for the study. A detailed history and thorough physical examination were followed by investigations like full blood count, blood grouping, serum urea, serum creatinine and buy Anlotinib random blood sugar. Patients were also screened for HIV infection using rapid test/ELISA test. A determination of CD 4 count was also performed in all HIV positive patients. Radiological investigations like X-ray abdomen erect and chest X-ray were done in all patients on the suspicion of diagnosis of perforated PUD. Other investigations included hematological profile, serum urea and electrolytes and urinalysis. The diagnosis of perforated

PUD was made from history, plain abdominal and chest radiographs, and confirmed at laparotomy. Patients were put on intra-venous fluids, nasogastric suction, intravenous antibiotics and intravenous

anti-ulcer drugs; adequate hydration was indicated by an hourly urine MLN2238 manufacturer output of 30 ml/hour. After adequate resuscitation, laparotomy was done through midline incision and identified the perforation site. Simple closure of the perforation and reinforcement with pedicled omental patch (Graham’s omentopexy) was done. Thorough peritoneal lavage with 3 to 4 liters of normal saline was followed by placement of intraperitoneal drain. The operations were performed either by a consultant surgeon or a senior resident under the direct supervision Etofibrate of a consultant surgeon. The Boey score [11] as a tool for outcome prediction was calculated based on data recorded at the time of admission to hospital. The Boey risk stratification in perforated peptic ulcer consists of associated medical illness, preoperative shock and long-standing perforation (more than 24 hours). Preoperative shock was defined as a preoperative systolic blood pressure of less than 90 mmHg. All the patients were put on triple regime consisting of Amoxicillin (500 mg TID), Metranidazole(400 mg TID) and Omeprazole (20 mg BID), all given orally for 14 days to eradicate H. Pylori. Patients were followed up on an out patient basis for up to 12 months after surgery.

Plasmids were extracted from overnight samples using QIAprep Spin

Plasmids were extracted from overnight samples using QIAprep Spin Mini Prep kit (Qiagen, Sussex, UK) according to the manufacturer’s instructions and sent for Sanger sequencing (Source BioSciences, Dublin, Ireland). Bioinformatic analysis Following Sanger sequencing, sequence

reads were analysed using the NCBI protein database (BlastX; (http://​blast.​ncbi.​nlm.​nih.​gov/​)). In the event where multiple hits occurred, the BLAST hit which displayed greatest homology is reported. Results and discussion A PCR-based approach highlights the presence of β-lactamase gene homologues in the gut microbiota The results of the β-lactamase-specific PCRs demonstrated the presence and diversity of class 2 β-lactamase genes in the gut microbiota of healthy adults (Table 2[32]). Of the β-lactam primers used, the primers designed find more to amplify bla TEM genes yielded the greatest number of unique sequence hits (42% of selected TOPO sub-clones gave a unique hit). The majority of these AG-881 nmr genes exhibited a high percentage identity with genes from various members of the Proteobacteria including E. coli, Klebsiella, Salmonella, Serratia, Vibrio parahaemolyticus and Escherichia vulneris. The resistance of PRIMA-1MET cell line strains of Salmonella and Serratia to β-lactams via bla TEM genes has been noted [33–35] and such strains have been associated with nosocomial infections [36]. In contrast, there have been relatively

few studies of bla TEM genes in Vibrio parahaemolyticus and Escherichia vulneris[37, 38]. The identification of genes homologous to those from Enterobacteriaceae is not surprising given the prevalence of resistance genes among

members of this family [12]. It was notable that the bla TEM primers also amplified genes that resembled bla TEM genes from some more unusual sources, including two genes from Baf-A1 clinical trial uncultured bacteria and from a Sar 86 cluster (a divergent lineage of γ-Proteobacteria) bacteria. This approach can thus provide an insight into possible novel/unusual sources of resistance genes, including those that culture-based approaches would fail to detect. Such results also highlight that had initial screening for resistant isolates been completed prior to PCR amplification of the resistance genes, such unusual sources of resistance genes may have been overlooked. Additionally, genes encoding ESBLs, including bla TEM-116, bla TEM-195 and bla TEM-96 amongst others, were also identified, with their closest homologues being members of the Proteobacteria (Table 2). Table 2 Homologues of β-lactamase genes detected in the human gut microbiota via PCR techniques Accession # Gene description Closest homologue E value % identity Bla TEM         ADE18890.1 β-lactamase TEM-1 S. enterica subsp. enterica 5e-154 99 AAS46844.1 β-lactamase TEM-1 S. marcescens 2e-156 100 AEN02824.1 β-lactamase TEM-1 K. pneumoniae 3e-111 99 AEN02817.

The AZO films AZO films with overall 1,090 cycles of ZnO plus Al2

The AZO films AZO films with overall 1,090 cycles of ZnO plus Al2O3 layers were alternatively deposited on quartz substrates

at 150°C. The ALD cycles in the ZnO/Al2O3 supercycles are 50/1, 22/1, 20/1, 18/1, 16/1, 14/1, 12/1, and 10/1, where monocycle Al2O3 doping layers were inserted between different cycles of ZnO sublayers. Since the real Smoothened Agonist in vitro Al U0126 solubility dmso concentration matches the ‘rule of mixtures’ formula well at lower Al concentration below 5%, in which the growth rate of the AZO is close to pure ZnO [19]. The Al concentration in the AZO films was calculated using the following formula: (1) where is the percentage of Al2O3 cycles, ρ Al, and ρ Zn are the densities of Al and Zn atoms deposited during each ALD cycle for the pure Al2O3 and ZnO films, respectively. The densities of Al2O3 and ZnO growth by ALD are 2.91 and 5.62 g/cm3[20], So ρ Al and ρ Zn were Selleck Tariquidar calculated to be 5.89 × 10−10 mol/cm2/cycle and 1.27 × 10−9 mol/cm2/cycle, respectively. Figure  3 shows the XRD patterns of the AZO films grown on quartz substrate with different ZnO/Al2O3 cycle ratios that are varied

from 50:1 to 10:1 (corresponding to Al concentration from 0.96% to 4.42%). The diffraction pattern of the pure ZnO film without Al2O3 doping layer is also shown as a reference. The X-ray diffraction pattern from pure ZnO film exhibits multiple crystalline ZnO structure with (100), (002), and (110) peaks [17]. With increasing the Al doping concentration, the (002) and (110) diffraction peaks decrease strongly, thus the AZO films exhibiting (100) dominated the orientation. The intensity of the (100) diffraction peak

reaches a maximum at 2.06% (with the ratio of ZnO/Al2O3 layers is 22/1), and then it decreases at Clostridium perfringens alpha toxin higher Al concentration above 3%. The preferred (100) orientation of the AZO films in our samples is consistent with the results reported by Banerjee et al. [18]. It is worthy to note that the Al2O3 layer by ALD is amorphous at the growth temperature of 150°C, so the decrease of the (100) peak at higher Al concentration can be explained that the amorphous Al2O3 doping layers destroy the crystal quality during the growth of AZO films. Figure  3 also shows that the (100) peak of ZnO shifts to larger diffraction angle with increasing the concentration of Al in AZO films. This can be interpreted as that the increase of the Al concentration will reduce the lattice constant by substitutions of Zn2+ ions (ion radius 0.74 Å) with smaller Al3+ (0.53 Å) ions; therefore, the (100) peak of ZnO shifts to larger diffraction angle in AZO films. Figure 3 XRD patterns of the AZO films with different Al content from 0% to 4.42%. Figure  4 plots the resistivity of AZO films as a function of Al concentration, which was measured by four-point probe technique. As the Al concentration increases from 0% to 2.26%, the resistivity initially decreases from 1.11 × 10−2 to a minimum of 2.38 × 10−3 Ω·cm, and then increases at higher Al doping concentration.

90%~99 70% and the deduced amino acid identities among them were

90%~99.70% and the deduced amino acid identities among them were 92.30%~100.00%, indicating that changes in amino acids were fewer than

those in nucleotides. The vp4s from 10 out of these 14 field strains of EV71 were also sequenced and analyzed with vp4s from other 22 strains of EV71 obtained from GenBank (see Additional file 1). The nucleotide identities in these vp4s were similar to those in vp1s but the deduced amino acid sequences for these vp4s were 98.60%~100.00%. In addition, nucleotide sequence comparisons BTK inhibitor cell line between sequences of EV71 isolated from mild cases and those of EV71 isolated from severe cases in the present study showed that there were no consistent divergences of nucleotides in vp1s or vp4s (data not shown). The vp1s from 14 strains of CA16 isolated from clinical specimens in this study were sequenced and analyzed with vp1s from 14 strains of CA16 obtained from GenBank (see Additional file 1). The nucleotide identities among them were 75.40% ~99.90% while the deduced amino acid identities of them were 91.20%~100.00%. The

nucleotide identities among those CA16 VP4s were 80.20%~100.00% and the deduced amino acids of them were identical (Table 1). Table 1 The nucleotide identities and amino acid identities for the corresponding genes Sequence name Number of strains Nucleotide identity (%) Amino acid identity (%) EV71 vp1s 35 80.90~99.70 click here 92.30~100.00 CA16 vp1s 28 75.40~99.90 91.20~100.00 EV71 vp1s/CA16 vp1s 35/28 62.00~66.80 70.00~72.70 EV71 vp4s 32 79.20~100.00 98.60~100.00 CA16 vp4s 15 80.20~100.00 100.00 EV71 vp4s/CA16 vp4s 32/15 64.30~73.90 78.30~79.70 The nucleotide sequences of vp1s between EV71 and CA16 were also compared using MegAlign of DNAStar. Both vp1 encoding gene from EV71 and CA16 was 891 nucleotides in length and the deduced amino acids of them were 297 in length. The identities of nucleotides for them were 62.50%~66.80% and the deduced amino acid identities for them were 70.00%~72.70%. The comparison between vp4s from EV71 and CA16 using MegAlign of DNAStar showed that the number of nucleotides was 207 in length and the

deduced amino acids of them were 69 in length. The identities of nucleotides among them were 64.30%~73.90% and the identities of the deduced amino acids were 78.30%~79.70% (Table 1). Phylogenetic analysis of complete vp1s and vp4s L-gulonolactone oxidase from EV71 and CA16 Phylogenetic analysis of EV71 was based on the alignment of complete vp1 and vp4 gene sequences from EV71. A total of 36 strains were included in the phylogenetic analysis of the EV71 vp1 genes. Among them, vp1s from 14 EV71 field strains were sequenced in this study, 8 strains available in GenBank were reported in other studies in China, 13 strains obtained from GenBank were used as genotype reference and CA16 strain G-10 was used as an outgroup. Phylogenetic analysis of complete EV71 vp1s showed that these 14 EV71 strains isolated in this study from 2007 to 2009 was closest to C4 sub-genotype (Figure 1A).

Iran J Environ Heal Sci Eng 2005,2(4):251–254 59 Rhoades JD: Sa

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of protein database search programs. Nuc Acid Res 1997,25(17):3389–3402.CrossRef 73. Thompson JD, Vildagliptin Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG: The CLUSTAL-X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucl Acid Res 1997,25(24):4876–4882.CrossRef 74. Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S: MEGA 5: molecular evolutionary genetic analysis using maximum likelihood, evolutionary distance and maximum parsimony methods. Mol Biol Evol 2011. doi:10.1093/molbev.msr121. Competing interests The authors declare that they have no competing interests. Authors’ contributions RCK, PC, LN and SS planned the study. PC performed the experiments. PC and RCK analyzed the results. RCK, PC, LN and SS drafted the manuscript.