(PDF 116 KB) Additional file 3: Table S3. Secreted proteins from Leishmania donovanii and their corresponding Trypanosoma orthologs. contains

the list of 358 proteins from L. donovanii identified in Silverman et al., 2008 [20] which were blasted against the T. brucei genome. The blast e scores > e-50 were reported as positive identification of T. brucei orthologs. Functional categories were assigned to L. donovanii-secreted proteins as well as the transmembrane span prediction (TMHMM) of these proteins. (PDF 28 KB) Additional file 4: Table S4. Proteins identified in glycosome from T. brucei [19]. contains the list of 163 proteins from the glycosome proteome which were classified into functional categories (MapMan bins nomenclature). (PDF 10 KB) Additional file 5: Table S5. Proteins identified in total proteome from T. brucei [18]. contains the list of 1071 proteins from the total proteome which were STA-9090 classified into functional categories (MapMan bins nomenclature). (PDF 40 KB) Additional file 6: Table S6. Genome-wide prediction of secreted proteins using SignalP and secretomeP. contains the list of 1445 SignalP-predicted proteins (containing a putative transit peptide) from T. brucei and classified according to the number of predicted transmembrane spans (TMHMM prediction) (sheet 1). SecretomeP-predicted proteins from T. brucei were reported

according to their p-value (sheet 2). The 3 highest classes p>0.9, 0.9>p>0.8, and 0.8>p>0.7 containing, respectively, 128, 583, and 875

proteins and their number of predicted transmembrane spans (TMHMM prediction) were reported. (PDF 119 KB) Additional Protein Tyrosine Kinase inhibitor file 7: Table S7. Proteins identified in sucrose fractionated membranes from infected rat serum (IRS). contains the list of the IRS proteins. IRS proteins shared with ESPs or exosome are boxed in yellow and orange, respectively. (PDF 9 KB) Additional file 8: Table S8. Additional informations on proteins identified in secretome. contains the list of the proteins identified in 1D and BN-PAGE gels spots. Protein score, number of peptides Terminal deoxynucleotidyl transferase identified and number of peptides that fit to our stringent filter are provided. (PDF 90 KB) References 1. Robinson NP, Burman N, Melville SE, Barry JD: Predominance of duplicative VSG gene conversion in antigenic variation in African trypanosomes. Mol Cell Biol 1999, 19:5839–46.PubMed 2. Dubois ME, Demick KP, Mansfield JM: Trypanosomes expressing a mosaic variant surface glycoprotein coat escape early detection by the immune system. Infect Immun 2005, 73:2690–7.PubMedCrossRef 3. MacGregor P, Matthews KR: Modelling trypanosome chronicity: VSG dynasties and parasite density. Trends Parasitol 2008, 24:1–4.PubMedCrossRef 4. WHO: Human African Trypanosomiasis (sleeping sickness): epidemiological update. Wkly Epidemiol Rec 2006, 81:71–80. 5. Stich A, Abel PM, Krishna S: Human African Trypanosomiasis.

3 × 105 S/cm) and the creation of new electrical contacts by nanowires. In the case of AgNWs alone, the AgNW/PVDF composites show no

percolation up to 2 vol % filler loading. By adding small amounts of TRGs (0.04 and 0.08 vol %), the hybrids display a steady increase in conductivity with increasing Ag content. Interestingly, the conductivity of AgNW/TRG/PVDF hybrids is much higher than the total Roscovitine solubility dmso conductivity of both TRG/PVDF and AgNW/PVDF composites. Thus, there exists a synergetic effect between these two types of nanofillers [42]. It seems that AgNWs can bridge the TRG sheets effectively, facilitating the transport of electrons among them [43]. The presence of conducting network can be detected by the alternating current (AC) response that manifested itself in a

conductivity plateau. Figure  3b shows the AC conductivity of PVDF filled with TRGs, AgNWs, and hybrid nanofillers. For the TRG/PVDF and AgNW/PVDF composites, electrical conductivity rises almost linearly with the frequency, click here implying these materials are insulators. In contrast, the conductivity of AgNW/TRG/PVDF composite is frequency independent from 102 to 107 Hz. This sample exhibits a DC conductivity plateau over a broad frequency range, showing the formation of good conducting network. Figure  3c is a schematic diagram illustrating the occurrence of synergistic effect between the AgNW and TRG fillers in a conductive network. On the contrary, the AgNW or TRG filler alone does not form a conducting path. The percolated AgNW/TRG/PVDF composite exhibits higher conductivity compared to a combined total conductivity of TRG/PVDF and AgNW/PVDF composites. From Figure  3a, the conductivity of 1 vol % AgNW/0.04 vol % TRG/PVDF hybrid is more than nine orders of magnitude higher than that of the 1 vol % AgNW/PVDF composite. Furthermore, the conductivity

of 2 vol % AgNW/0.08 vol % TRG/PVDF, i.e., 10 S/cm is comparable to that of measured graphite paper with a conductivity of 12 S/cm [44]. Figure  4a,b is the SEM micrographs showing typical morphologies of hybrid composites. The AgNWs are well dispersed within the polymer matrix. The use of sonication during the composite Ponatinib fabrication process can reduce the aspect ratio of AgNWs as expected.The effect of temperature (40 to 180°C) on electrical resistivity (a reciprocal of conductivity) of AgNW/TRG/PVDF hybrids is now discussed (Figure  5). All hybrid composites show a slow increase in resistivity with increasing temperature initially followed by a sharp increase in resistivity as the temperature approaches melting point of PVDF. This behavior is commonly referred to as the positive temperature coefficient (PTC) effect of resistivity. A maximum increase in resistivity is particularly apparent for the composite with 0.04 vol % TRG and 1 vol % AgNW loadings, being more than four orders of magnitude higher than that at 40°C. Above the melting temperature of PVDF, a reverse effect, i.e.

Virus Res. 2008;132:257–62.PubMed 101. Jiang J-H, Wang N, Li A, Liao W-T, Pan Z-G, Mai S-J, et al. Hypoxia can contribute to the induction of the Epstein–Barr virus (EBV) lytic cycle. SCH772984 J Clin Virol. 2006;37:98–103.PubMed 102. Keely S, Glover LE, Weissmueller T, MacManus CF, Fillon S, Fennimore B, et al. Hypoxia-inducible factor-dependent regulation of platelet-activating factor receptor as a route for Gram-positive bacterial translocation across epithelia. Mol Biol Cell.

2010;21:538–46.PubMedCentralPubMed 103. Spear W, Chan D, Coppens I, Johnson RS, Giaccia A, Blader IJ. The host cell transcription factor hypoxia-inducible factor 1 is required for Toxoplasma gondii growth and survival at physiological oxygen Tyrosine Kinase Inhibitor Library high throughput levels. Cell Microbiol. 2006;8:339–52.PubMed 104. Wiley M, Sweeney KR, Chan DA, Brown KM, McMurtrey C, Howard EW, et al. Toxoplasma gondii activates hypoxia-inducible factor (HIF) by stabilizing the HIF-1α subunit via type I activin-like receptor kinase receptor signaling. J Biol Chem. 2010;285:26852–60.PubMedCentralPubMed 105. Degrossoli A, Bosetto MC, Lima CBC, Giorgio S. Expression of hypoxia-inducible factor 1α in mononuclear phagocytes infected with Leishmania amazonensis. Immunol Lett. 2007;114:119–25.PubMed

106. Singh AK, Mukhopadhyay C, Biswas S, Singh VK, Mukhopadhyay CK. Intracellular pathogen Leishmania donovani activates hypoxia inducible factor-1 by dual mechanism for survival advantage within macrophage. PLoS ONE. 2012;7:e38489.PubMedCentralPubMed 107. Zhao S, Wu J. Hypoxia inducible factor stabilization as a novel strategy to treat anemia. Curr Med Chem. 2013;20:2697–711.PubMed 108. Peyssonnaux C, Cejudo-Martin P, Doedens A, Zinkernagel AS, Johnson RS, Nizet V. Cutting edge: essential role of hypoxia inducible factor-1α in development of lipopolysaccharide-induced sepsis. J Immunol. 2007;178:7516–9.PubMed 109. Glycogen branching enzyme Thiel M, Caldwell CC, Kreth S, Kuboki S, Chen P, Smith P, et al. Targeted deletion of HIF-1α gene in T cells prevents their inhibition in hypoxic inflamed tissues and improves septic mice survival. PLoS ONE. 2007;2:e853.PubMedCentralPubMed

110. Schafer ST, Frede S, Winning S, Bick A, Roshangar P, Fandrey J, et al. Hypoxia-inducible factor and target gene expression are decreased in patients with sepsis: prospective observational clinical and cellular studies. Anesthesiology. 2013;118:1426–36.PubMed 111. Keely S, Campbell EL, Baird AW, Hansbro PM, Shalwitz RA, Kotsakis A, et al. Contribution of epithelial innate immunity to systemic protection afforded by prolyl hydroxylase inhibition in murine colitis. Mucosal Immunol. 2014;7:114–23.PubMed 112. Campbell EL, Bruyninckx WJ, Kelly CJ, Glover LE, McNamee EN, Bowers BE, et al. Transmigrating neutrophils shape the mucosal microenvironment through localized oxygen depletion to influence resolution of inflammation. Immunity. 2014;40:66–77.PubMed 113. Weigert A, Weichand B, Sekar D, Sha W, Hahn C, Mora J, et al.

The most similar genus is Botryosphaeria. Cophinforma has morphologically unique ascospores which are hyaline and aseptate. Generic type: Cophinforma eucalypti signaling pathway Doilom, J.K. Liu & K.D. Hyde. Cophinforma eucalypti Doilom, J.K. Liu & K.D. Hyde., sp. nov. MycoBank: MB 801316 (Fig. 16) Fig. 16 Cophinforma eucalyptus (MFLU 12–0752, holotype) a-b. Ascostromata on dead twigs of Eucalyptus sp. c. Ascostromata cut horizontally showing the white contents. d–e. Vertical section through ascostromata. f. Immature asci and mature asci. g. Immature ascus. h–j. Asci. k–m. Ascospores. n. Germinating ascospore. Scale bars: d–e = 100 μm, f = 50 μm, g–j, n = 20 μm, k–m = 10 μm

Etymology: Referring to the host “Eucalyptus sp.,” on which the fungus was collected. Saprobic

on recently fallen wood. Ascostromata (88-)112–125(−130) μm high × (135-)172–185(−195) μm wide \( \left( \overline x = 112 \times 165\,\upmu \mathrmm,\mathrmn = 10 \right) \), initially immersed under host epidermis, becoming semi-immersed to erumpent, breaking through cracks in bark, gregarious and fused, uniloculate, globose to subglobose, membraneous, visible white contents distinct when cut, ostiolate. Ostiole (33-)43–52 μm high, (31-)40–48 μm wide, central, papillate, pale brown, relatively broad, periphysate. Peridium (13-) 28–34 μm wide, broader at the base, comprising several layers of relatively think-walled, dark brown

to black-walled cells arranged in a textura angularis. Pseudoparaphyses XL184 hyphae-like, numerous, embedded in a gelatinous matrix. Asci 74–90(−123) × 17–23 μm \( \left( \overline x = 89 \times 20\,\upmu \mathrmm,\mathrmn = 10 \right) \), 8-spored, bitunicate, fissitunicate, clavate to cylindro-clavate, sometimes short pedicellate, mostly long pedicellate, apex rounded with an ocular chamber. Ascospores 21–26 × 8–11 μm \( \left( \overline x = 23.5 \times 9\,\upmu \mathrmm,\mathrmn = 20 \right) \), overlapping uniseriate to biseriate, hyaline, aseptate, ellipsoidal to obovoid, slightly wide above the centre, minutely guttulate, smooth-walled. 17-DMAG (Alvespimycin) HCl Asexual state not established. Culture characteristics: Ascospores germinating on PDA within 8–15 h. Germ tubes produced from both ends of the ascospore. Colonies growing on PDA 80 mm diam after 3 d at 25–30 °C, fast growing; fimbriate, flat or effuse, dense, initially white after a few days becoming pale grey starting form the centre, finally dark grey to black, convex with papillate surface, reaching the edge the Petri dish after 4 d. Material examined: THAILAND, Chiang Rai Province, Muang District, Thasud Sub District, on dead branch of Eucalyptus sp., 5 October 2011, M. Doilom, (MFLU 12–0752, holotype), ex-type living culture MFLUCC 11–0425; Ibid., living culture MFLUCC 11–0655. Lasiodiplodia Ellis & Everh., Bot. Gaz.

J Bacteriol 2007,189(6):2540–2552.PubMedCentralPubMedCrossRef 54. Spratt BG, Maiden MC: Bacterial population genetics, evolution and epidemiology. Philos Trans R Soc Lond B Biol Sci 1999,354(1384):701–710.PubMedCentralPubMedCrossRef AZD2014 mw 55. Thompson FL, Iida T, Swings J: Biodiversity of vibrios. Microbiol Mol Biol Rev 2004,68(3):403–431.PubMedCentralPubMedCrossRef Competing interests The authors declare that they have no competing interests. Author’s contributions SU did the experimental design, performed the experiments, analyzed the data and drafted the manuscript. TA and SH participated in study design, data analysis and drafting the manuscript. GG participated

in selection of strains and drafting the manuscript. MK, LS and UM took part in preparing and performing the experiments. All authors have read and approved the manuscript.”
“Background Hospital Acquired Infections (HAI) have exacted a heavy toll worldwide with over 2 million patients annually contracting an infection in the US [1], being one of the leading causes of death in the US behind cancer and strokes [2]. In Europe, out of 3 million HAI [3] approximately

50,000 resulted in death [4], and in Australia more than 177,000 HAI occur per year [5] whilst in the click here province of Quebec, Canada the rate of HAI are estimated to be around 11% [6]. The HAI rates in developing countries are significantly higher [7–9]. According to the USA Center for Disease Control (CDC) some of the predominant HAI organisms are Staphylococcus aureus, Pseudomonas aeruginosa, and Enterobacter species [10]. Methicillin resistant S. aureus accounts for 50% of HAI associated with multidrug resistant pathogens [10]. The Extended Prevalence of Infection in Intensive Care (EPIC II) study demonstrated a 50% HAI

rate in ICU patients sampled from over 75 countries and two of the most predominant organisms were resistant Staphylococci and BCKDHA P. aeruginosa[11]. HAI are associated with considerable mortality, morbidity and costs [2, 12]. Recent intervention efforts including improvement of national surveillance, use of aggressive antibiotic control programs, healthcare staff education for improved hygiene, isolation of infected patients, use of disposable equipment, cleaning and disinfection of environmental surfaces and equipment, improvement of cleaning equipment and sanitary facilities, increase in nursing and janitorial resources and better nutrition [13–17], have been shown to reduce HAI rates. However further supplemental interventions are required. The link between contaminated hard surfaces to HAI has been demonstrated [18–28] and an antimicrobial protected touch surface would assist in reduction of pathogen buildup upon touch surfaces as long as that activity can be indisputably demonstrated.

A small chelate constant (lg β) would benefit the combination of F- and Ln3+ ions resulting in the NaLuF4 lattice [28]. According to coordination chemistry, the chelate constants increase for sodium citrate, SDS, DDBAC, and PEG according to priority [27], resulting in gradually increasing size of UCNPs. Another reason may be attributed to the diverse viscosity of interface of dual phase system after adding surfactant [29]. Figure 1 TEM image of (a) ILs-UCNPs, (b,c) Cit-UCNPs. Figure 2 SEM images of (a) SDS-UCNPs, (b) DDBAC-UCNPs, and (c) PEG-UCNPs. To evaluate the ligand stability in each sample, TGA was performed (Additional file 1: Figures S1c, S2c,

ABT-199 molecular weight S3c, S4c, S5c). TGA curves showed two weight loss stages in the range of 20°C to 900°C. The first weight loss stage in the temperature range of 20°C to 200°C was due to the loss of absorbed water. The second stage from 200°C to 900°C was attributed to the combustion of the organic groups in the samples. A common feature was that weight

of each sample decreased rapidly at 600°C to 700°C. Additionally, when temperature reached 600°C, the weight loss was still less than 10% of the total weight, indicating good stability of each ligand linking. Notably, Cit-Na had shown priority in chelate ability, whose weight loss was only 1.82% until temperature risen up to 900°C. Based on EDX spectrums (Additional file 1: Figures S1d, S2d, S3d, S4d, S5d), fluorine had occupied majority RG7204 ic50 weight of UCNPs, demonstrating that the lead role of capping agent was still ILs, and other surfactants worked as cooperative assistants to develop functional surface. The successful ligand links between surfactants and surface of UCNPs were further verified by FTIR spectroscopy. Figure 3 showed the FTIR spectra of the five UCNP samples. The transmission band peaks at approximately 2,930 this website and 2,854 cm-1 can be assigned to the asymmetric and symmetric stretching vibrations, respectively. However, these features were lost in the spectrum of the Cit-UCNPs sample, suggesting

the disappearance of the –CH2-CH2– groups. What is more, bands peaks at 1,641 and 1,520 cm-1 belonged to the C = O vibrations, indicating the presence of carboxylic groups in Cit-UCNPs. Band peak at 1,206 cm-1 in Figure 3 (c) suggested that the sulfonic acid groups have been attached in the surface. In Figure 3 (d), band peaks at 2,924, 1,532, and 749 cm-1 indicate the presence of phenyl group. Peak at 1,524 cm-1 in Figure 3 (e) could indicate new groups had been attached. On the basis of the above described FTIR results, it can be deduced that the active groups of surfactants capped successfully onto UCNP surface during the synthetic process though part of surface still linked with long alkyl chains from ILs. As a consequence, ILs and surfactants participate synthesis process together as capping agents, competing with each other to cap for UCNPs.

Karaman MW, Herrgard S, Treiber DK, Gallant P, Atteridge CE, Campbell BT, Chan KW, Ciceri P, Davis MI, Edeen PT, Faraoni R, Floyd M, Hunt JP, Lockhart DJ, Milanov ZV, Morrison MJ, Pallares G, Patel HK, Pritchard S, Wodicka LM, Zarrinkar PP: A quantitative analysis of kinase inhibitor selectivity. Nat Biotechnol 2008, 26: 127–132.CrossRefPubMed click here 14. Di Leo A, Moretti E: Anthracyclines: the first generation of cytotoxic targeted agents? A possible dream. J Clin Oncol 2008, 26: 5011–5013.CrossRefPubMed

15. Llovet JM, Ricci S, Mazzaferro V, Hilgard P, Gane E, Blanc JF, de Oliveira AC, Santoro A, Raoul JL, Forner A, Schwartz M, Porta C, Zeuzem S, Bolondi L, Greten TF, Galle PR, Seitz JF, Borbath I, Haussinger D, Giannaris T, Shan M, Moscovici M, Voliotis D, Bruix J: Sorafenib in advanced hepatocellular carcinoma. N Engl J Med 2008, 359: 378–390.CrossRefPubMed 16. Motzer RJ, Hutson TE, Tomczak P, Michaelson MD, Bukowski RM, Rixe O, Oudard S, Negrier S, Szczylik C, Kim ST, Chen I, Bycott PW, Smoothened antagonist Baum CM, Figlin RA: Sunitinib versus interferon alfa in metastatic renal-cell

carcinoma. N Engl J Med 2007, 356: 115–124.CrossRefPubMed 17. Freidlin B, Simon R: Adaptive signature design: an adaptive clinical trial design for generating and prospectively testing a gene expression signature for sensitive patients. Clin Cancer Res 2005, 11: 7872–7878.CrossRefPubMed 18. Paz-Ares L, Sanchez JM, Garcia-Velasco A, Massuti B, Lopez-Vivanco G, Provencio M, Montes A, Isla D, Amador ML, Rosell R, G Spanish Lung Cancer: Monoiodotyrosine A prospective phase II trial of erlotinib in advanced non-small cell lung cancer (NSCLC) patients (p) with mutations in the tyrosine kinase (TK) domain of the epidermal growth factor receptor (EGFR). J Clin Oncol (Meeting Abstracts) 2006, 24: 7020. 19. El-Maraghi RH, Eisenhauer EA: Review of phase II trial designs used in studies of molecular targeted agents: outcomes and predictors of success

in phase III. J Clin Oncol 2008, 26: 1346–1354.CrossRefPubMed 20. Ratain MJ, Glassman RH: Biomarkers in phase I oncology trials: signal, noise, or expensive distraction? Clin Cancer Res 2007, 13: 6545–6548.CrossRefPubMed 21. Stone A, Wheeler C, Barge A: Improving the design of phase II trials of cytostatic anticancer agents. Contemp Clin Trials 2007, 28: 138–145.CrossRefPubMed 22. Kopec JA, Willison KD: A comparative review of four preference-weighted measures of health-related quality of life. J Clin Epidemiol 2003, 56: 317–325.CrossRefPubMed 23. Rosner GL, Stadler W, Ratain MJ: Randomized discontinuation design: application to cytostatic antineoplastic agents. J Clin Oncol 2002, 20: 4478–4484.CrossRefPubMed 24. Karapetis CS, Khambata-Ford S, Jonker DJ, O’Callaghan CJ, Tu D, Tebbutt NC, Simes RJ, Chalchal H, Shapiro JD, Robitaille S, Price TJ, Shepherd L, Au HJ, Langer C, Moore MJ, Zalcberg JR: K-ras mutations and benefit from cetuximab in advanced colorectal cancer. N Engl J Med 2008, 359: 1757–1765.CrossRefPubMed 25.

6 U of TrueStart Taq DNA polymerase (Fermentas, Lithauen) and 10 μM of both oligos in a 20 μl volume was performed. The program consisted of activation step at 95 °C

for 3 min and 5 cycles of denaturation at 95 °C for 30 s, annealing at 55 °C for 30 s and extension at 72 °C for 15 s. Final extension was 15 min at 72 °C. Template Ponatinib in vivo oligo sequences are listed in Additional file 3. Ninety-six templates were divided into four pools and each pool was tested separately with all of the probes on the microarray. Ligation reaction Ligation reactions were carried out in a 10 μl volume containing 1X Pfu ligase buffer (Agilent Technologies, Santa Clara, CA, USA), herring sperm DNA (Sigma-Aldrich, Steinheim, Germany), 30 mM tetramethylammonium chloride (TMAC; Sigma-Aldrich, Steinheim, Germany), about 200 ng of environmental template DNA, 400 amol of each probe and 2 U of Pfu ligase (Agilent Technologies, Santa Clara, CA, USA). The reaction was LDE225 nmr cycled for 20 rounds at 94 °C for 30 s and at 56 °C for 8 min in a thermal cycler (MJ Research, MA, USA). PCR from ligated probes The PCR reaction mixture for amplification of circularised ligation products contained 1X Paq HS buffer (Agilent Technologies, Santa Clara,

CA, USA), 200 μM of each dNTP, 0.5 μM forward primer (5′-Cy3-CGACGTTGTAAAACGACGGCCAGT-3′), 0.5 μM reverse primer (5′-phosphate-TTTCACACAGGAAACAGCTATGAC-3′), 2.5 U of Paq5000 DNA polymerase (Agilent Technologies, Santa Clara, CA, USA) and 10 μl of ligation reaction in a final volume of 30 μl. The PCR program consisted of activation step at 95 °C for 3 min and 35 cycles of denaturation

at 95 °C Exoribonuclease for 20 s, annealing at 58 °C for 14 s and extension at 72 °C for 5 s. The PCRs were done in Arktik thermal cycler (Finnzymes, Espoo, Finland) with block-mode temperature control using manufacturer’s PCR tubes. Microarrays The microarray experiments were performed on Arrayit or Agilent microarray platforms. The 16 compartment slides purchased from Arrayit (Sunnyvale, CA, USA) were designed and used as described previously [42]. Briefly, for hybridisation to Arrayit microarrays, a mixture containing 20 μl of PCR/lambda exonuclease reaction, 5X SSC, 20 μg of herring sperm DNA (Sigma-Aldrich, Steinheim, Germany) and 5 pmol of control oligo in a final volume of 60 μl was applied to each subarray according to manufacturer’s instructions. The hybridisation was carried out in the dark at 55 °C for 2 h. After hybridisation, the microarray was washed for 3X15 min in 0.1X SSC, 0.1% SDS and briefly with water. Finally, the slide was air dried. The high-density custom oligo microarrays were manufactured by Agilent (Santa Clara, CA, USA) in 8 X 15 K format. Each of eight subarrays contained 1500 cZipCode oligos in ten replicates. Hybridisation to Agilent microarrays was performed according to manufacturer’s instructions.

The explorative approach of this study, combined with the finding of a decreased intracellular DCFH2 oxidation, revealed

an additional stimulation of cellular antioxidative mechanisms when exposed to CMH. This may contribute to an improved performance through increased ability to cope with training-induced Belnacasan nmr increases in oxidative stress. Combined effects of increased energy load and improved antioxidative defences may thus be the key to the performance improvement experienced by some athletes following creatine supplement, but this approach needs further investigation [41]. Acknowledgements The authors wish to thank Hanne S. Møller, Inge Lise Sørensen and Anne-Grete Dyrvig Petersen for excellent technical assistance. The Danish Technology and Production Research Council (FTP) is thanked for financial support through the project “”NMR-based metabonomics on tissues and biofluids”" (project no. 274-05-339). The Danish National Research Foundation and the Danish Biotechnological Instrument Centre (DABIC) is acknowledged. References 1. Balsom PD, Söderlund K, Ekblom B: Creatine in humans with special reference to creatine supplementation. Sports Med 1994, 18:268–280.CrossRefPubMed 2. Casey A, Constantin-Teodosiu D, Howell S, Hultman E, Greenhaff PL: Creatine ingestion favorably affects performance AG-014699 in vitro and muscle metabolism

during maximal exercise in humans. Am J Physiol 1996, 271:E31-E37.PubMed 3. Dentowski ABH, Opaszowski D, Blachnio D, Polanowski B: Effect of creatine supplementation

on the performance in supramaximal, intermittent exercise. Biol Sport 1997, 14:291–298. 4. Mujika I, Padilla S: Creatine supplementation as an ergogenic acid for sports performance in highly trained athletes: a critical review. Int J Sports Med 1997, 18:491–496.CrossRefPubMed 5. Gotshalk LA, Kraemer WJ, Mendonca MAG, Vingren JL, Kenny AM, Spiering BA, Hatfield DL, Fragala MS, Volek JS: Creatine supplementation improves muscular performance in older women. Eur J Appl Physiol 2008, 102:223–231.CrossRefPubMed 6. Balsom PD, Söderlund K, Sjödin B, Ekblom B: Skeletal muscle 17-DMAG (Alvespimycin) HCl metabolism during short duration high-intensity exercise: influence of creatine supplementation. Acta Physiol Scand 1995, 154:303–310.CrossRefPubMed 7. Burke DG, Candow DG, Chilibeck PD, MacNeil LG, Roy BD, Tarnopolsky MA, Ziegenfuss T: Effect of creatine supplementation and resistance-exercise training on muscle insulin-like growth factor in young adults. Int J Sport Nutr Exerc Metab 2008, 18:389–398.CrossRefPubMed 8. Safdar A, Yardley NJ, Snow R, Melov S, Tarnopolsky MA: Global and targeted gene expression and protein content in skeletal muscle of young men following short-term creatine monohydrate supplementation. Physiol Genomics 2008, 32:219–228.PubMed 9. McMorris T, Mielcarz G, Harris RC, Swain JP, Howard A: Creatine supplementation and cognitive performance in elderly individuals.

Moreover,

since other next generation sequencing platforms will allow a greater sequencing depth, this may allow a deeper characterization of the microbial community and could reveal additional differences in the microbial community composition for the various conditions measured in this study. Finally, our study also reveals that microbial disruption by bead-beating allows greater detection of Gram-positive bacteria such as Blautia (Firmicutes phylum) and Bifidobacterium (Actinobacteria phylum), commonly detected in human PI3K inhibitor stools. In conclusion, the hydration of faecal samples and their degree of homogenisation do not significantly alter their microbial community composition and structure. However, although the mechanical disruption of microbial cells causes genomic DNA degradation in simulated diarrhoeic stool samples, our findings confirm that this step is necessary for the detection of Gram-positive bacteria such as Blautia and Bifidobacterium. Methods Ethics statement Subjects provided their written consent to participate see more in this study, and the Institutional Review Board of the Vall d’Hebron Hospital (Barcelona, Spain)

approved this consent procedure. Sample collection protocol Stools were collected from eight healthy participants. The collection protocol involved providing participants with an ice bag containing an emesis basin (Ref. 104AA200, PRIM S.A, Spain), a 50-mL sterile sampling bottle (Ref. 409526.1, Deltalab, Spain), a sterile spatula (Ref. 441142.2, Deltalab, Spain), and gloves (Additional file very 3: Figure S2) during their visit to the laboratory. For the purpose of stool collection, the participants were instructed to do the following once at home: 1) use the emesis basin to collect the stool; 2) after the deposit, transfer it to the sampling bottle ensuring no homogenisation; 3) take it to the lab within the first 3 hours after deposit; and 4) in the laboratory, the samples were processed as mentioned in the experimental design, and then the samples were stored at -80°C. Naming convention Since

the samples from same individuals were used to test different factors that could affect microbial composition, a labeling nomenclature had to be settled down as indicated in Table 1. The “D” stands for “diarrhoea” in the water content study. The “L” stands for “layer”, “O” for “outer”" and “I” for the “inner”" layer of the stool, and “H” for “homogenised stool” in the homogenisation evaluation. The “P” stands for samples that contained PBS to simulate diarrhoea not undergoing bead-beating, while “B” stands for samples that did not contain PBS, but underwent bead-beating. Samples with the “C” label are controls that did not contain PBS and did not undergo bead-beating. The numbers 1–8 signify the 8 different volunteers. Genomic DNA extraction To evaluate the need for stool homogenisation during collection, aliquots (250 mg) of each sample were suspended in 0.1 M Tris (pH 7.