Control: the cells treated with C. butyricum. Discussions The intestinal epithelial cell surface represents the largest exposed surface of the body that must be protected by the immune system against toxic substance and pathogenic bacteria. All intestinal epithelial cells are usually capable of regulating the immune response through different mechanisms,

one of which is the secretion of anti-inflammatory cytokines. buy PRN1371 Throughout the present study, we have focused on the role of IL-10 in regulating epithelial cell function. IL-10 is a potent GSK126 inhibitor of pro-inflammatory cytokine production, and has been shown to inhibit production of IL-6 and IL-1β in macrophages [18, 19]. Supporting evidence for a role for IL-10 in inflammation is derived from studies in mice deficient in IL-10 or harboring mutated IL-10, which are a model of enterocolitis [20]. These IL-10−/− mice under normal conditions show increased inflammatory responses and develop inflammatory bowel disease. Moreover, these IL-10−/− mice are extremely susceptible to infection-induced immunopathology [21]. All these data suggest that endogenous IL-10 synthesis plays an important role in vivo in down-regulating immune responses and preventing host immunopathology. Moreover, beneficial effects

in colitis patients have been obtained via probiotic bacteria-induced IL-10 production [22]. In our current study, C. butyricum stimulates elevated levels of IL-10 in HT-29 cells. Because this website this probiotic strain is frequently used in the management of allergic diseases or gastroenteritis, it is hypothesized that it promotes mucosal tolerance mediated through

IL-10. Therefore, we further assessed the role of IL-10 in probiotic-mediated immune modulation by neutralizing or knocking down IL-10 in HT-29 cells. It was found that disruption of IL-10 enhanced effects of C. butyricum-induced NF-κB activation and IL-8 secretion. The results demonstrate that C. butyricum modifies the mucosal immune response to modulate the levels of specific molecules such as cytokines by increasing IL-10 levels and consequently decreasing inflammatory cytokines. The viability of cells is dependent on cytokines. However, high-dose cytokines can induce apoptosis and necrosis. Bacteria and their metabolites can induce an anti-proliferative effect through induction of apoptosis [23–25]. Fluorometholone Acetate In the current study, disruption of IL-10 enhanced C. butyricum-induced IL-8 secretion. We further assessed whether this probiotic strain induced apoptosis and necrosis of HT-29 cells due to a lack of effect of IL-10. The results showed that the number of abnormal cells significantly increased compared to the control, indicating that disruption of IL-10 caused a loss of suppression of the mucosal immune response and even excessive apoptosis and necrosis. This study confirmed that C. butyricum exerts anti-inflammatory effects and enhances tolerance to bacteria through increasing IL-10 production.

Colour development was monitored at 450 nm in a multiwell plate reader (Thermo Fisher Scientific, Shangai). Caspase-3 activity evaluation Caspase-3 activity was determined in leukemia cells using a colorimetric kit from Biovision (Milpitas, CA, USA) in accordance with the manufacturer’s

instructions. The assay is based on the spectrophotometric detection at 405 nm of the chromophore p-nitroaniline (pNA) after cleavage from the labeled substrate DEVD-pNA by caspase-3. Protein concentration in the cytosolic extracts was measured using the Bradford method [24]. DNA fragmentation analysis The genomic DNA fragmentation was evaluated by agarose gel electrophoresis of DNA isolates obtained by the salting-out method [25]. For this purpose, leukemia cells were grown in the presence or absence of CF 5 μl/ml up to 72 h; a positive control (cells treated for 6 h with 25 μM etoposide) was also included. After counting signaling pathway and washing, cells were subjected to DNA extraction. The DNA samples were carefully resuspended in TE buffer; the nucleic acid concentration and purity were measured using a NanoDrop® ND-1000 spectrophotometer (Thermo-Scientific,

Wilminton, DE, USA). 2 μg of each sample was loaded onto 1.5% TAE agarose gel; DNA laddering was visualized on a UV transilluminator by ethidium bromide staining. Images were obtained using a Gel Doc 2000 (Bio-Rad Laboratories S.r.l, Segrate, MI, Italy). HIF-1α measurement HIF-1α quantification was performed in leukemia cells using an enzyme-linked immunosorbent assay kit from Abcam (Cambridge, UK), in accordance with the manufacturer’s STI571 clinical trial instructions. Colour development was evaluated at 450 nm in a multiwell plate reader (Thermo

Fisher Scientific, Docetaxel chemical structure Shangai). Protein concentration in cell extracts was measured using the Bradford method [24]. Western blot assay of BKM120 research buy GLUT-1 Leukemia cells were grown in presence or absence of CF 5 μl/ml up to 72 h. After counting and washing, cells were resuspended in 1X SDS loading buffer to 20×106 cells/ml. Cell lysis was achieved by vortex, and the viscosity was reduced by passing through a syringe needle. 15 μl of each samples were run on 0.8% SDS-polyacrylamide gel and the resolved proteins were electrophoretically transferred to supported nitrocellulose membranes (Bio-Rad Laboratories S.r.l, Segrate, MI, Italy) using a Bio-Rad Semidry Transfer system. Non-specific binding to membranes was blocked by incubation in blocking solution (50 mM Tris–HCl, 150 mM NaCl and 5% (w/v) non-fat dried milk, pH 7.5). After blocking solution removal, membranes were incubated in a new blocking solution with a rabbit polyclonal GLUT-1 antibody (PA1-46152, Thermo Scientific) at 4°C overnight. Membranes were then washed three times with TTBS (50 mM Tris–HCl, 150 mM NaCl and 0.05% (v/v) Tween 20, pH 7.

Passlick B, Pantel K, Kubuschok B, Angstwurm M, Neher A, Thetter O, Schweiberer L, Izbicki JR: Expression of MHC molecules and ICAM-1 on non-small cell lung carcinomas: association with early lymphatic spread of tumour cells. Eur J CP673451 Cancer 1996, 32A:141–145.PubMed 26. Vitale M, Rezzani R, Rodella L, Zauli G, Grigolato P, Cadei M, Hicklin DJ, Ferrone S: HLA class I antigen and

transporter associated with antigen processing (TAP1 and TAP2) down-regulation in high-grade primary breast carcinoma lesions. Cancer Res 1998, 58:737–742.PubMed 27. Saio M, Teicher M, Campbell G, Feiner H, Delgado Y, Frey AB: Immunocytochemical demonstration of down regulation of HLA class-I molecule expression in human metastatic breast carcinoma. Clin Exp Metastasis 2004, 21:243–249.PubMed 28. Ryschich E, Notzel T, Hinz U, Autschbach F, Ferguson SBE-��-CD mw J, Simon I, Weitz J, Frohlich B, Klar E, Buchler MW, Schmidt J: Control of T-cell-mediated immune response by HLA class I in human pancreatic carcinoma. Clin Cancer Res 2005,11(2 Pt 1):498–504.PubMed 29. Sharpe JC, Abel PD, Gilbertson JA, Brawn P, Foster CS: Modulated expression of human leucocyte antigen class I and class II determinants in hyperplastic and malignant human selleck inhibitor prostatic epithelium. Br J Urol 1994, 74:609–616.PubMed 30. Brasanac D, Markovic-Lipkovski J, Hadzi-Djokic J, Muller GA, Muller CA: Immunohistochemical analysis of HLA class II antigens and tumor infiltrating mononuclear cells in renal cell carcinoma:

correlation with clinical and histopathological data. Neoplasma 1999, 46:173–178.PubMed 31. Hilders CG, Houbiers JG, van Ravenswaay Claasen HH, Veldhuizen RW, Fleuren GJ: Association between HLA-expression and infiltration of immune cells in cervical carcinoma. Lab Invest 1993, 69:651–659.PubMed 32. Hilders CG, Munoz IM, Nooyen Y, Fleuren GJ: Altered HLA expression by metastatic cervical carcinoma cells as a factor in impaired immune surveillance. Gynecol Oncol 1995, 57:366–375.PubMed 33. Cruz I, Meijer CJ, Walboomers JM, Snijders PJ, Van der Waal I: Lack of MHC class I surface expression on neoplastic cells and poor activation of the secretory pathway of cytotoxic cells in oral squamous cell carcinomas.

Br J Cancer 1999, 81:881–889.PubMed 34. Grandis JR, Falkner DM, Melhem MF, Gooding WE, Drenning SD, Morel Oxalosuccinic acid PA: Human leukocyte antigen class I allelic and haplotype loss in squamous cell carcinoma of the head and neck: clinical and immunogenetic consequences. Clin Cancer Res 2000, 6:2794–2802.PubMed 35. Gati A, Da Rocha S, Guerra N, Escudier B, Moretta A, Chouaib S, Angevin E, Caignard A: Analysis of the natural killer mediated immune response in metastatic renal cell carcinoma patients. Int J Cancer 2004, 109:393–401.PubMed 36. Lanier LL: Natural killer cells: from no receptors to too many. Immunity 1997, 6:371–378.PubMed 37. Doubrovina ES, Doubrovin MM, Vider E, Sisson RB, O’Reilly RJ, Dupont B, Vyas YM: Evasion from NK cell immunity by MHC class I chain-related molecules expressing colon adenocarcinoma.

Physiol Plant 100:214–223 Asada K, Heber U, Schreiber U (1992) Pool size of electrons that can be donated to P700+ as determined in intact leaves: donation to P700+ from stromal components via the intersystem chain. Plant Cell Physiol 33:927–932 Bailey S, Walters RG, Jansson S, Horton P (2001) Acclimation of Arabidopsis thaliana to the light environment: the existence of separate low light and high light responses. Planta 213:794–801PubMed Bailey S, Horton P, Walters RG (2004) Acclimation

of Arabidopsis thaliana to the light environment: the relationship between photosynthetic function and chloroplast composition. Planta 218:793–802PubMed Bilger W, Björkman O (1990) Role of the xanthophyll cycle in photoprotection elucidated by measurements of light-induced absorbance changes, fluorescence and photosynthesis in Selleck 4-Hydroxytamoxifen leaves of Hedera canariensis. EPZ5676 molecular weight Photosynth Res 25:173–185PubMed Björkman O, Demmig

B (1987) Photon yield of O2 evolution and chlorophyll fluorescence Alpelisib cell line characteristics at 77 K among vascular plant of diverse origins. Planta 170:489–504PubMed Bradbury M, Baker NR (1981) Analysis of the slow phases of the in vivo chlorophyll fluorescence induction curve. Biochim Biophys Acta 63:542–551 Bradbury M, Baker NR (1984) A quantitative determination of photochemical and non-photochemical quenching during the slow phase of the chlorophyll fluorescence induction curve of bean leaves. Biochim Biophys Acta 765:275–281 Brestic M, Zivcak M (2013) PSII fluorescence techniques for measurement of drought and high temperature stress signal in crop plants: protocols and applications. In: Rout GR, Das AB (eds) Molecular stress physiology of plants. Springer, Berlin Brestic M, Cornic G, Fryer M, Baker N (1995) Does photorespiration protect the photosynthetic apparatus in French bean leaves from photoinhibition during drought stress? Planta 196:450–457 Brestic M, Zivcak M, Kalaji MH, Allakhverdiev SI, Carpentier R (2012) Photosystem II thermo-stability in situ: environmentally

induced acclimation and genotype-specific reactions in Triticum aestivum L. Plant Physiol Biochem 57:93–105PubMed Briantais JM, Merkelo H, Govindjee (1972) Lifetime of the excited state τ in vivo. III. Chlorophyll Glutathione peroxidase during fluorescence induction in Chlorella pyrenoidosa. Photosynthetica 6:133–141 Bussotti F, Desotgiu R, Cascio C, Pollastrini M, Gravano E, Gerosa G, Marzuoli R, Nali C, Lorenzini G, Salvatori E, Manes F et al (2011) Ozone stress in woody plants assessed with chlorophyll a fluorescence. A critical reassessment of existing data. Environ Exp Bot 73:19–30 Butler WL (1978) Energy distribution in the photochemical apparatus of photosynthesis. Annu Rev Plant Physiol 29:345–378 Cascio C, Schaub M, Novak K, Desotgui R, Bussotti F, Strasser RJ (2010) Foliar responses to ozone of Fagus sylvatica L. seedlings grown in shaded and in full sunlight conditions.

In this study, we have characterized the effect

of the MgFnr protein on growth and magnetite biomineralization in MSR-1. Deletion of Mgfnr did not affect the growth yield, but impaired magnetosome formation under microaerobic conditions only in the presence of nitrate (i.e., when denitrification was active) but not in its absence. This implies that MgFnr might be involved in magnetite synthesis by regulation of denitrification genes, whereas check details expression of terminal oxidases for O2 respiration is likely not under the control of MgFnr, similar to Fnr from Shewanella oneidensis[33]. In fact, we found that neither the rates of oxygen consumption nor transcription of terminal oxidase genes [34] displayed any difference between the WT and ΔMgfnr mutant. The presence of putative Fnr binding sites in the promoter regions of all operons of denitrification further indicates that MgFnr is involved in controlling the transcription of denitrification genes in response to different oxygen concentrations. Consistent with this, transcription patterns of denitrification genes in ΔMgfnr mutant were different from WT. For example, in the ΔMgfnr strain the expression of nap was no longer GM6001 cost upregulated by oxygen, expression of nirS was much higher under aerobic conditions than WT, and aerobic expression of nor and nosZ was no longer repressed but upregulated by oxygen. Furthermore,

we failed to identify a putative Fnr protein encoded in the genome of the nondenitrifying magnetotactic bacteria Magnetococcus marinus or Desulfovibrio magneticus strain RS-1, which also suggests that Histone Methyltransferase inhibitor & PRMT inhibitor Fnr of MTB is likely only responsible to regulate genes encoding for denitrification, but not required for aerobic respiration. In addition, we also observed significantly decreased N2 evolution in deep slush agar tubes in ΔMgfnr mutant. This raised the question at which step(s) of

denitrification is affected by the loss of MgFnr. We propose that this is Sclareol not likely caused by the reduction steps from NO3 – to N2O based on the following observations: (i) The consumption rate of NO3 – and NO2 – did not decrease in ΔMgfnr mutant; (ii) NO is lethal to the cells while no defective growth was found in ΔMgfnr mutant, and no NO emission was observed during mass spectrometry experiments which also implies that the activity of NO reductase is not decreased; (iii) The N2O emission rate after addition of nitrate was similar for ΔMgfnr mutant and WT. Therefore, we conclude that loss of MgFnr affects the last step of denitrification, the reduction of N2O to N2. In agreement, the emission rate of N2 was lower for ΔMgfnr mutant than for the WT. However, we cannot exclude the possibility that loss of MgFnr has an impact on further pathways involved in biomineralization other than denitrification.

). Images were analyzed with Fingerprinting II Informatix software (Version 3.0, Bio-Rad). Band matching and cluster analysis was performed using an unweighted pair group method with arithmetic averages (UPGMA) and the Dice coefficient with 1% optimization and tolerance levels. Based on the dendrogram obtained from the cluster analysis, letters were assigned

to designate fla types and numbers were assigned to designate PFGE types. Isolates with > 90% similarity were assigned to the same fla type or PFGE type. GSK2118436 purchase composite cluster analysis including fla typing, PFGE, and antimicrobial resistance testing selleck inhibitor data was performed using the Fingerprinting II Informatix software. The composite dendrogram was determined by UPGMA using the average from

the experiment as a coefficient for similarity and correction for internal weights. Statistical analysis The χ2 test was used to analyze the significance of the difference between ciprofloxacin and erythromycin resistance rates, including C. jejuni compared to C. coli in each plant, and pre chill compared to post chill in plant A. An α of 0.01 was used for statistical significance. The discriminatory ability of fla typing, PFGE, antimicrobial resistance profiling, and composite analysis was calculated using the numerical index of discrimination (D) according to the method of Hunter and Gaston [60]. The discriminatory index represents the probability that two unrelated strains sampled from the test population will be placed into different typing groups [60]. Acknowledgements The authors gratefully acknowledge the see more U.S. this website Food and Drug Administration for financial and technical assistance. We also thank Curt Doetkott, North Dakota State University (NDSU), for statistical consultation and Dr. Mohamed Fakhr, University of Tulsa, for assistance with data analysis and manuscript review. References 1. Mead PS, Slutsker L, Dietz V, McCaig LF, Bresee JS, Shapiro C, Griffin PM, Tauxe RV: Food-related illness and death in the United States. Emerg Infect Dis 1999, 5:607–625.CrossRefPubMed 2. Butzler JP:Campylobacter , from

obscurity to celebrity. Clin Microbiol Infect 2004, 10:868–876.CrossRefPubMed 3. Allos BM:Campylobacter jejuni infections: update on emerging issues and trends. Clin Infect Dis 2001, 32:1201–1206.CrossRefPubMed 4. Jacobs-Reitsma W:Campylobacter in the food supply. Campylobacter American Society for Microbiology, Washington, D.C 2 Edition (Edited by: Nachamkin I, Blaser MJ). 2000, 467–481. 5. Cox NA, Stern NJ, Craven SE, Berrang ME, Musgrove MT: Prevalence of Campylobacter and Salmonella in the cecal droppings of turkeys during production. J Appl Poult Res 2000, 9:542–545. 6. Luangtongkum T, Morishita TY, Ison AJ, Huang S, McDermott PF, Zhang Q: Effect of conventional and organic production practices on the prevalence and antimicrobial resistance of Campylobacter spp. in poultry.

In: Bell SS, McCoy ED, Mushinsky HR (eds) Habitat structure: the physical arrangement of objects in space. Chapman and Hall, London Bohnsack JA, Eklund AM, Szmant AM (1997) Artificial reef research: is there more than the attraction-production issue? Fisheries 22:14–16 Bortone SA (1998) Resolving the attraction-production dilemma in artificial reef research: some yeas and nays. Fisheries 23:6–10CrossRef Brattegard T, Holthe T (1997) Distribution of marine, benthic macro-organisms in Norway. Research report for DN 1997-1. Directorate for Nature Managment Bros WE (1987) Effects of removing or adding Caspase activity assay structure (Barnacle Shells) on recruitment

to a fouling community in Tampa-Bay, Florida. J Exp Mar Biol Ecol 105:275–296CrossRef Buckley LB et al (2010) Phylogeny, niche conservatism and the latitudinal diversity gradient in mammals. Proc Royal Soc B 277:2131–2138CrossRef HDAC inhibitor Coull BC, Wells JBC (1983) Refuges from fish predation: experiments with phytal meiofauna from the New Zealand rocky intertidal. Ecology 64:1599–1609CrossRef Dean T (1981) Structural aspects of sessile invertebrates as organizing forces in an estuarine fouling community. J Exp Mar Biol Ecol 53:163–180CrossRef Dipper F (1991) Colonisation and natural changes in a newly established ‘artificial reef’ in Wnt activation Gulf waters. In: Elliott M, Ducrotoy J-P (eds) Estuaries and coasts: spatial and temporal intercomparisons. Olsen and Olsen, University of Caen Eilertsen

HC, Taasen JP (1984) Investigations on the plankton community of Balsfjorden, northern Norway. The phytoplankton 1976–1978. Environmental factors, Phosphoglycerate kinase dynamics of growth, and primary production. Sarsia 69:1–15 Faulkner GH (1930) The anatomy and the histology of bud-formation in the Serpulid Filograna implexa, together with some cytological observations on the nuclei of the neoblasts. J Linn Soc (Zool) XXXVII:109–191CrossRef Fischer AG (1960) Latitudinal variations in organic diversity. Evolution 14:64–81CrossRef Freiwald A et al (2004) Cold-water coral reefs.

UNEP-WCMC, Cambridge Gaarder KR (1938) Phytoplankton studies from the Tromsø district 1930–1931. Yearbooks of Tromsø Museum 55:1–159 Gabriele M et al (1999) Sublittoral hard substrate communities of the northern Adriatic Sea. Cah Biol Mar 40:65–76 Gaston KJ (1996) Biodiversity––latitudinal gradients. Prog Phys Geogr 20:466–476CrossRef Gaston KJ (2000) Global patterns in biodiversity. Nature 405:220–227PubMedCrossRef Gray JS (2001) Marine diversity: the paradigms in patterns of species richness examined. Sci Mar 65:41–56CrossRef Gulliksen B, Sandnes O (1980) Marine bunndyrsamfunn, “nøkkelarter” og felteksperimenter på hardbunn (In Norwegian). Fauna 33:1–9 Haines JL, Maurer D (1980a) Quantitative faunal associates of the Serpulid polychaete Hydroides dianthus. Mar Biol 56:43–47CrossRef Haines JL, Maurer D (1980b) Benthic invertebrates associated with a Serpulid polychaete assemblage in a temperate estuary.

A difference between F4 and F5/F6 is that the core-shell structures of the latter can be clearly seen in the CB-839 solubility dmso projection of the core from the shell. This is thought phosphatase inhibitor library to be associated with the increase of drug content, which makes the nanofibers brittle. The higher contents of quercetin in the shell of fibers F5 and F6 made them easier to fracture, and thus the core projects a little from the shell after breaking. TEM images of fibers F2, F4, F5, and F6 are shown in Figure 5. The uniform contrast of F2 suggests that the quercetin is distributed in the EC matrix at the molecular level, with no aggregates (Figure 5a). Fibers F4, F5, and F6 have evident core-shell structures (Figure 5b,c,d).

Except for the heterogeneous region in the shell of F6 (see Figure 5d), no nanoparticles were observed in the three core-shell fibers, indicating uniform structures. The heterogeneous region in Figure 5d may be the result of a migration of the core components to the shell, or phase separation may have happened within the shell due to the high quercetin content in F6. Figure 5 TEM images. (a) F2, (b) F4, (c) F5, and (d) F6. Physical state of quercetin XRD analyses were conducted to determine the physical status of

the drug in the nanofibers. Quercetin, a yellowish green powder to the naked eye, comprises polychromatic crystals in STA-9090 clinical trial the form of prisms or needles. The crystals exhibit a rough surface under cross-polarized light (Figure 6a). The data in Figure 6b show the presence of numerous distinct Bragg reflections in the XRD pattern of pure quercetin, demonstrating

its existence as a crystalline material. The PVP and EC diffraction patterns Adenosine exhibit a diffuse background with two diffraction haloes, showing that the polymers are amorphous. The patterns of fibers F2, F4, F5, and F6 show no Bragg reflections, instead consisting of diffuse haloes. Hence, the composite nanofibers are amorphous, and quercetin is not present as a crystalline material in the fibers. Figure 6 Physical form investigation. (a) Crystals of quercetin viewed under cross-polarized light and (b) XRD patterns of the raw materials and nanofibers. These results concur with the SEM and TEM observations. No crystalline features are observed for any of the nanofibres. The heterogeneous region in Figure 5d is thus thought unlikely to be because of the recrystallization of quercetin, but most probably this anomaly comprises a composite of the drug and PVP with a higher concentration of quercetin than its surroundings. In vitro drug release profiles The in vitro drug release profiles of the four different nanofibers are given in Figure 7. As anticipated, the monolithic nanofibers F2 (containing only quercetin and EC) exhibited a sustained release profile as a result of the poor water solubility of quercetin and the insolubility of EC. In contrast, the core-shell fibers F4, F5, and F6 showed an initial burst release of 31.7%, 47.2%, and 56.

Therefore, we assumed that

measuring changes in foot volumes using plethysmography was an accurate method as well. A limitation in our study is the fact that we did not determine total body water as it has been reported in studies investigating changes in total body water during exercise for example through the diluted isotope method [42, 43]. This might provide more insight into the hydration status in ultra-marathoners, since we can only assume that total body water was increased in the slower runners leading to peripheral oedemas in these subjects. Furthermore, we did not ask our athletes about wearing compression stockings [47]. Elastic compression stockings can prevent the development of oedema in long-haul

flights [48]. It would be interesting to determine in future field-studies, whether compression stockings have an influence on the development of peripheral check details oedemas in ultra-marathoners. The foot swelling might also be a high protein interstitial space fluid swelling and may be associated with markers of skeletal muscle damage. Leg swelling might also be due to venous insufficiency with a higher prevalence at advanced ages [49]. However, when plotting changes in foot volume versus age, we found no association between changes in foot volume and an increase in age (Figure 10). Figure 10 The change in the volume of the right foot was not associated with the age of the subjects ( r = 0.01, p = 0.91). Conclusions In summary, this study demonstrated that fluid intake was positively related to the volume of the foot in 100-km ultra-marathoners. https://www.selleckchem.com/products/Belinostat.html An increase in the foot volume

occurred in athletes with an increased fluid intake. In addition, slower running speed was associated Vildagliptin with an increase in the foot volume and the change in foot volume was negatively correlated to the change in plasma [Na+]. Therefore, we concluded that fluid overload occurred in slower runners and was responsible for the development of oedemas in the foot. In addition, post-race plasma [Na+] decreased in those runners. Our data support the finding that fluid overload is the main risk factor for developing EAH [19–21]. For AZD9291 clinical trial practical application, athletes performing an ultra-marathon should be aware that excessive drinking with fluid overload increases the risk for EAH [19–21] and can lead to the development of peripheral oedemas in the foot. Acknowledgements The authors thank the race director of ’100 km Lauf Biel’ for his support to perform this study. We are in great debt to the athletes who enabled us for the data collection. References 1. Knechtle B, Senn O, Imoberdorf R, Joleska I, Wirth A, Knechtle P, Rosemann T: Maintained total body water content and serum sodium concentrations despite body mass loss in female ultra-runners drinking ad libitum during a 100 km race. Asia Pac J Clin Nutr 2010, 19:83–90.PubMed 2.

aeruginosa is a frequently isolated bacterium Gamma-secretase inhibitor that causes septicemia and death [17]. It is a ubiquitous opportunistic, non-fermenting, gram-negative rod that can infect patients with impaired immune systems. Treatment ofP. aeruginosa

infection is frequently hindered by antibiotic resistance, and multi-drug resistant find more strains are mostly isolated from burn wound infections [3,4,20]. An efficient vaccine is therefore needed. After colonizing the site of the burn,P. aeruginosa produces several virulence factors, such as exotoxin A, alkaline protease and elastase, which affect the host tissue. High titers of antitoxin against exotoxin A in patients infected withP. aeruginosa reduces the risk septicemia and death [9,21]. Table 3 Survival rates, presence of exotoxin A, culture results and colony counts in the experimental group (immunized mice) inoculated withP. aeruginosa

Post-inoculation time (day) Number of animals alive (survival rate, %) CFU/mL from inoculated burns Exotoxin A in sera (%)* Positive culture (%) Number of animals alive (survival rate, %)           Liver Spleen Blood 1 48 (100) 1.5 × 108 ND 48 (100) – - – 4 48 (100) 1.4 × 107 ND 48 (100) – - – 7 47 (98) 1.3 × 106 ND 47 (100) 1 (2) 1 (2) 1 (2) 11 46 (96) 1.2 × 105 ND 47 (98) 1 (2) RG7112 1 (2) 1 (2) 14 45 (94) 1 × 104 ND 45 (94) 1 (2) 1 (2) 1 (2) ND, not detectable by CIEP; * neutralizing antibody detected Conclusion Exotoxin A is the principal lethal factor ofP. aeruginosa. It seems logical that a toxoid of exotoxin A could be used as an effective vaccine. Our study shows that in mice immunized with semi-purified exotoxin Fossariinae A, a protective titer of antitoxin developed that effectively prevented the experimentally infected animals from septicemia and death. The majority (93.8%) of immunized infected mice survived

during 70 days of observation after a burn wound was inoculated withP. aeruginosa while all the non-immunized mice in the control group died. The rising antibody titer in the surviving mice and the decrease in the mortality rate indicate the presence of an effective antitoxin in the immunized mice. Pavlovskis et al. [22] found that the survival rate did not increase significantly following active immunization with a toxoid of exotoxin A and infection withP. aeruginosa in burned mice. However, Matsumato et al. [5] found that immunization with a combination of alkaline protease and toxoid of exotoxin A decreased mortality. Some investigators have reported that active immunization with a lipopolysaccharide and an outer membrane protein (OMP) ofP. aeruginosa could control the infection in the burned area [23,24]. Our study, using a semi-purified exotoxin A that contained trace amounts of LPS and OMP, points to a higher efficacy than a toxoid prepared from purified exotoxin A.