J Exp Med Selleckchem Belnacasan 2002, 195:415–422.PubMedCrossRef 18. Zhong W, Gern L, Stehle T, Museteanu C, Kramer M, Wallich R, Simon MM: Resolution of experimental and tick-borne Borrelia burgdorferi infection in mice by passive, but not active immunization using recombinant OspC. Eur J Immunol 1999, 29:946–957.PubMedCrossRef 19. Hodzic E, Feng S, Freet KJ, Borjesson

DL, Barthold SW: Borrelia burgdorferi population kinetics and selected gene expression at the host-vector interface. Infect Immun 2002, 70:3382–3388.PubMedCrossRef 20. Salazar CA, Rothemich M, Drouin EE, Glickstein L, Steere AC: Human Lyme arthritis and the immunoglobulin G antibody response to the 37-kilodalton arthritis-related protein of Borrelia burgdorferi . Infect Immun 2005, 73:2951–2957.PubMedCrossRef 21. Tunev SS, Hastey CJ, Hodzic E, Feng S, Barthold SW, Baumgarth N: Lymphadenopathy during Lyme borreliosis is caused by spirochete migration- induced specific B cell activation. PLoS Pathog 2011, 7:e1002066.PubMedCrossRef 22. Hodzic E, Feng S, Freet K, Barthold SW: Borrelia burgdorferi population dynamics and prototype gene expression during infection of immunocompetent and immunodeficient mice. Infect Immun 2003, 71:5042–5055.PubMedCrossRef 23. Liang FT, Yan J, Mbow ML, Sviat SL, Gilmore RD, Mamula M, Fikrig E: Borrelia burgdorferi changes its surface antigenic expression in response to host immune responses. Infect Immun 2004, see more 72:5759–5767.PubMedCrossRef

24. Probert WS, LeFebvre RB: Protection of C3H/HeN mice from challenge with Borrelia burgdorferi through active immunization with OspA, OspB, or OspC, but not with OspD or the 83-kilodalton

antigen. Infect Immun 1994, 62:1920–1926.PubMed 25. Bankhead T, Chaconas G: The role of VlsE antigenic variation in the Lyme disease spirochete: persistence through a mechanism that differs from other pathogens. Mol Microbiol 2007, 65:1547–1558.PubMedCrossRef 26. Labandeira-Rey M, Seshu J, Skare J: The absence of linear plasmid 25 or 28–1 of Borrelia burgdorferi dramatically alters the kinetics of experimental infection via distinct mechanisms. Infect Immun 2003, 71:4608–4613.PubMedCrossRef 27. Labandeira-Rey M, Skare JT: Decreased infectivity in Borrelia burgdorferi strain B31 is associated with 17-DMAG (Alvespimycin) HCl loss of linear plasmid 25 or 28–1. Infect Immun 2001, 69:446–455.PubMedCrossRef 28. Purser JE, Norris SJ: Correlation between plasmid content and infectivity of Borrelia burgdorferi . Proc Natl Acad Sci USA 2000, 97:13865–13870.PubMedCrossRef 29. Xu Q, Seemanapalli SV, Lomax L, McShan K, Li X, Fikrig E, Liang FT: Association of linear plasmid 28–1 with an arthritic phenotype of Borrelia burgdorferi . Infect Immun 2005, 73:7208–7215.PubMedCrossRef 30. Pal U, Wang P, Bao F, Yang X, Samanta S, Schoen R, Wormser GP, Schwartz I, Fikrig E: Borrelia burgdorferi basic membrane proteins A and B participate in the genesis of Lyme arthritis. J Exp Med 2008, 205:133–141.PubMedCrossRef 31.

Secondary antibody conjugated to horseradish

peroxidase was obtained from Bio-Rad. Visualisation was done by the enhanced chemiluminescent reaction (Stratagene). Non-denaturating PAGE was performed using 7.5% (w/v) polyacrylamide gels pH 8.5 and included 0.1% (w/v) Triton-X100 in the gels [14]. Samples (25 μg of protein) were incubated with 5% (v/v) Triton X-100 prior to application to the gels. Where indicated, the relative intensity Acalabrutinib of hydrogenase staining and protein amount from immunoblots was quantified using ImageJ from the National Institutes of Health [36]. Hydrogenase activity-staining was done as described in [14] except that the buffer used was 50 mM MOPS pH 7.0. Acknowledgements We are grateful to Nadine Taudte and Gregor Grass for supplying strains and the plasmid pFEO and to Frank Sargent for supplying anti-hydrogenase antisera. This work was supported by a grant from the Deutsche Forschungsgemeinschaft (DFG SA494/3-1). Electronic supplementary

material Additional file 1: Plasmid-encoded FeoB synthesis in MC4100 and PM06 ( feoB ::Tn 5 ). Extracts (25 μg protein in membrane sample buffer) from Selleck GDC-973 MC4100 and PM06, transformed with pECD1079 bearing feoB and pFEO bearing the whole feo operon, both cloned behind a tetracycline promotor and encoding an N-terminal StrepII-tag on FeoB encoded on pECD1079 were separated by SDS-PAGE (10% w/v polyacrylamide) and after transfer to nitrocellulose detected by incubation with Strep-tactin conjugated to horseradish peroxidase. Strains were grown either with or without aeration in TGYEP, pH 6.5 and

gene expression was induced with 0.2 μg ml-1 AHT (anhydrotetracycline) as indicated. Biotin carboxyl carrier protein (BCCP) served as a loading control. The sizes of the protein standards are shown on the right side of the gel. The angled arrow indicates the position of the Strep-FeoB polypeptide. Extracts Amino acid derived from MC4100 and PM06 transformed with pFEO did not synthesize Strep-tagged FeoB and therefore acted as a negative control. (TIFF 371 KB) References 1. Vignais P, Billoud B: Occurrence, classification, and biological function of hydrogenases: an overview. Chem Rev 2007, 4206–4272. 2. Forzi L, Sawers RG: Maturation of [NiFe]-hydrogenases in Escherichia coli . Biometals 2007, 20:565–578.PubMedCrossRef 3. Pinske C, Krüger S, Soboh B, Ihling C, Kuhns M, Braussemann M, Jaroschinsky M, Sauer C, Sargent F, Sinz A, Sawers RG: Efficient electron transfer from hydrogen to benzyl viologen by the [NiFe]-hydrogenases of Escherichia coli is dependent on the coexpression of the iron-sulfur cluster-containing small subunit. Arch Microbiol 2011, in press. 4. Lukey MJ, Parkin A, Roessler MM, Murphy BJ, Harmer J, Palmer T, Sargent F, Armstrong FA: How Escherichia coli is equipped to oxidize hydrogen under different redox conditions. J Biol Chem 2010, 285:3928–3938.PubMedCrossRef 5. Böck A, King P, Blokesch M, Posewitz M: Maturation of hydrogenases. Adv Microb Physiol 2006, 51:1–71.

P. fluorescens is a widespread gram-negative bacterium present in a variety of ecological niches such as refrigerated food products, soil, water [5] and in the digestive tract [6]. Interestingly, a highly specific antigen of P. fluorescens, designated as I2, was detected in the serum of 54% of the patients suffering from ileal Crohn’s disease (CD) [7] and a direct link between the severity of the pathology and the level of circulating I2 antigen has been demonstrated https://www.selleckchem.com/EGFR(HER).html [8]. Surprisingly, the proinflammatory potential of this bacterium or its interaction with the intestinal epithelium has never been investigated. Several studies have focused on the mucosal immune response to pathogenic bacteria.

selleck products Human IECs infected with

pathogenic bacteria generally produce proinflammatory cytokines, such as interleukin (IL)-8 [9]. The latter has a chemotactic role and can recruit polymorphonuclear cells into the infected site and promote their infiltration of the epithelial layer infected by invasive or noninvasive bacteria [10, 11]. IL-8 gene expression is regulated by two major transcriptional factors: nuclear factor kappa B (NF-κB) and activator protein (AP)-1 [12]. NF-κB has a pivotal role in the immune and inflammatory response, but also controls cell survival, proliferation and differentiation [13, 14]. Recent works demonstrated that NF-κB signaling is a critical element of the homeostatic immuno-inflammatory function in the gut. Indeed, epithelial NF-κB preserves the integrity of the gut epithelial barrier and coordinates the antimicrobial actions

of the innate and adaptive immune systems [15]. Nevertheless, hyperactivation of this transcription factor results in chronic inflammatory bowel diseases [16]. Activation of AP-1 is dependent on mitogen-activated protein kinases (MAPK) that are central in many physiological processes, including regulation of cytokine and stress responses and cytoskeletal reorganization [17, 18]. P. fluorescens MFN1032 is a clinical strain recently isolated in our laboratory [19]. It displays hemolytic activity toward sheep erythrocytes [20, 21], however, its infectious potential on human IECs is still unknown. In the present study, we investigated adhesion Metalloexopeptidase and cytotoxic properties of P. fluorescens MFN1032 on Caco-2/TC7 and HT-29 cell lines in comparison to the psychrotrophic strain, P. fluorescens MF37 and the well-known opportunist pathogen P. aeruginosa PAO1. The proinflammatory potential of P. fluorescens MFN1032 was also evaluated by the measurement of IL-8 secretion on both Caco-2/TC7 and HT-29 cells, and analysis of NF-κB and AP-1 activation using the reporter gene strategy. Results Adhesion to intestinal epithelial cells The binding index of the clinical strain P. fluorescens MFN1032 on Caco-2/TC7 and HT-29 cells was determined after 5 h of incubation and compared to P. fluorescens MF37 and P. aeruginosa PAO1.

19) FOR No specified conception on project level   FOR investigated the effects of climate

change on Swiss forests. To the question “is there any sort of forest ideal that would play a role in the project?” it was stated: “For our project not really. Well I think people that see the forest as a working forest will probably have their visions of how the STAT inhibitor forest should best look like. But for the project, it’s not really, it doesn’t play a big role” (FOR 1, p. 10) POLL Environment–development combination: sustainable land use in the Indian Kodagu region stands for a functioning, diverse landscape, containing enough natural areas for conserving biodiversity while providing important (pollination) ecosystem services for productive agricultural systems A1 (A2), B2 Biodiversity conservation and its potential benefit to crop production were at the core of the project’s underlying notion of sustainable land use, which was embedded in the greater vision “to manage the landscape in a manner that is delivering not just secure livelihoods for the people who are living Rucaparib supplier in this area, but also securing the well-being of the (…) biodiversity and the land cover, but also the esthetics of the

landscape” (POLL 2, p. 4) LIV Environment–development combination: a more sustainable development in the Madagascan Manompana corridor comprises local people using the forests (i.e., its products) without clearing them, and using the cleared agricultural land efficiently so that food production is sufficient. The context offers well-regulated land rights and income generating alternatives to agriculture. A minimal level of wellbeing is reached, replacing acute poverty A1, A3, B1, B2 LIV’s sustainability conception concerned a region with rapid forest decline, characterized by subsistence economy and acute poverty among local people. The interviewee added to this rather concrete vision: “my goal is actually to shape the agricultural planning in such a way that in all these different aspects,

as little as possible changes to the negative for the local population. And at the same time for the forest” (LIV, p. 12/13) PALM Environment–development combination: In the investigated Indonesian region, (-)-p-Bromotetramisole Oxalate a sustainable development contains an oilpalm development that allows local smallholders to reach and maintain a decent standard of living in a self-determined way, and at the same time preserves the forests A1 (A3), B1, B4 PALM was concerned with oil palm development in Indonesia. With regard to core characteristics of a sustainable land use, the interviewee said: “I think it has to be something that you can support by itself. So it’s not something that relies too much on outside inputs. It can support by itself and people and not, what do you call that? “propertied” people and not make poor because of it.

Instead, this pathophysiological effect may be restricted to infections displaying a relevant liver involvement. Further work is still necessary to define the full impact of infections in FGF15/19 function and to determine the underlying molecular mechanisms. Conclusions Through the alteration of the hepatobiliary function, bacterial pathogens of the enterohepatic system dysregulate the homeostasis of the FGF15/19-FGFR4 endocrine axis. These revealing findings have important implications for the understanding of the pathophysiology of microbial diseases.

Disruption of the FGF15/19-FGFR4 pathway may be a contributing factor to the metabolic and nutritional disorders associated with infectious diseases. Acknowledgments We thank Catherine Desrosiers, Melisange selleck compound Roux and Elora Midavaine for technical help. This work was supported by grants to A.M. from the Fonds de Recherche du Québec-Santé (26710) and the Natural Sciences and Engineering Research Council of Canada (401949–2011), and to B.B.F. from the Canadian Institutes for Health

Research. L. C. M. A. was funded by a postdoctoral fellowship from the Canadian Institutes of Health Research. A. M. is a member of the FRQS-funded Centre de Recherche Clinique Étienne-Le Bel. References 1. Powanda MC, Beisel WR: Metabolic effects of infection on protein and energy status. J Nutr 2003,133(1):322S-327S.PubMed 2. McGuinness OP: Defective glucose homeostasis during infection. Annu Rev Nutr 2005, 25:9–35.PubMedCrossRef 3. Khosla SN: Typhoyd fever. Its cause, transmission and prevention. New Delhi: Atlantic www.selleckchem.com/products/chir-99021-ct99021-hcl.html Publishers; 2008. 4. Antunes LC, Arena ET, Menendez A, Han J, Ferreira RB, Buckner MM, Lolic P, Madilao LL, Bohlmann J, Borchers CH, et al.: Impact of salmonella infection on host hormone metabolism revealed by metabolomics. Infect Immun 2011,79(4):1759–1769.PubMedCrossRef 5. Parry CM: Epidemiological and clinical aspects of human typhoid fever. In Salmonella infections:

clinical, immunological IMP dehydrogenase and molecular aspects. Edited by: Mastroeni P, Maskell D. Cambridge, New York: Cambridge University Press; 2006. 6. Inagaki T, Choi M, Moschetta A, Peng L, Cummins CL, McDonald JG, Luo G, Jones SA, Goodwin B, Richardson JA, et al.: Fibroblast growth factor 15 functions as an enterohepatic signal to regulate bile acid homeostasis. Cell Metab 2005,2(4):217–225.PubMedCrossRef 7. Jones SA: Physiology of FGF15/19. In Endocrine FGFs and Klothos. Edited by: Kuro-o M. New York: Landes Bioscience and Springer Science; 2012:171–182.CrossRef 8. Potthoff MJ, Kliewer SA, Mangelsdorf DJ: Endocrine fibroblast growth factors 15/19 and 21: from feast to famine. Genes Dev 2012,26(4):312–324.PubMedCrossRef 9. Chiang JY: Bile acids: regulation of synthesis. J Lipid Res 2009,50(10):1955–1966.PubMedCrossRef 10.

III Number of study patients not indicated; mistletoe group included 155 patients. IV Numbers given only Decitabine cost for mistletoe group. V Not applicable for retrolective studies. Table 3 Controlled Clinical Studies on VAE Treatment in Breast and Gynaecological Cancer: Survival Site Stage Intervention (evaluable patients) Survival Outcomes Author,

year, reference       Years (median) Hazard ratio 5-year survival and others P-value 95% CI   Randomized controlled trials Breast T1a-3, N0, M0 Iscador (38) 14.8 0.65   0.2 0.34–1.25 Grossarth 2006a [52, 53, 135]     None (38) 13.8             IIIA–IIIB Iscador (17) 6.3 0.46   0.13 0.16–1.31 Grossarth 2001a [59, 135, 166]     None (17) 2.3             T1-3, N0-3, M0, local recurrence Surgery, radiationI, Helixor (192) Not applicableII   69.1% 5-year survival 0.048   Gutsch 1988 [62]     Surgery, radiationI,

CMF (177)     67.7% 5-year survival 0.025         Surgery, radiationI (274)     59.7% 5-year survival       Breast, others All stages Iscador (39) 3.5 (mean)     0.04   Grossarth 2001b [59]     None (39) 2.5 (mean)           Cervix IVA-B Iscador (19) 1.83 0.46   0.12 0.18–1.21 Grossarth 2007c [51]     None (19) 1.92           Uterus IA-C Iscador (30) 6.29 0.36   0.014 0.16–0.82 Grossarth 2008a [49]     None (30) 5.17             IVA-B Iscador (26) 1.5 1   0.99 0.46–2.16 Grossarth Rapamycin ic50 2008b [49] 3-mercaptopyruvate sulfurtransferase     None

(26) 2.0           Ovary IA–IC Iscador (21) 6.75 0.40   0.058 0.15–1.03 Grossarth 2007a [50]     None (21) 5.58             IV Iscador (20) 2.75 0.33   0.033 0.12–0.92 Grossarth 2007b [50]     None (20) 1.58           Non-randomized controlled studies Breast T1-3, N0, M0 Iscador (84)III 11.75 0.42   0.0002 0.27–0.68 Grossarth 2006b [52, 53, 135]     None (84) 10.13             Local recurrence, N0, M0 Iscador (29)IV 5.17     0.0025   Grossarth 2001b [59, 135]     None (29) 4.33             T1-4, N>1, M0 Iscador (38)IV 4.04     0.0516   Ø same study     None (38) 3.17             TX, NX, M1 Iscador (53)IV 3.08     0.0056   Ø same study     None (53) 2.17             I–III Iscador, (76)     29% alive 1985, after 11–14 years not shown   Salzer 1987 [66]     Radiation, hormone (79)     24% alive 1985, after 11–14 years       Cervix IB-IVA Iscador (102)III 7.17 0.41   <0.0001 0.27–0.63 Grossarth 2007f [51]     None (102) 5.92             IV Iscador (66)III 2.33 0.54   0.015 0.32–0.89 Grossarth 2007g [51]     None (66) 1.

Following the approach of Schubert et al. [31] we detected comparable ratios of ITS signal/mycelial biomass at different this website levels of fungal mycelium. In contrast, with another approach Raidl et al. [30] quantified the ITS copy number of P. croceum by using Taqman PCRs and by measuring the extent of mycelium from thin layers of sterile mycelium. To conclude, we could here clearly demonstrate how specific qPCR assays can be a powerful tool for elucidating the relative fungal and bacterial biomass in microcosm samples of varying complexity. Promotion of AcH 505 growth by P. croceum and response to soil microbial community P. croceum promotes AcH 505

growth, which may indicate that the MHB feeds on fungal exudates. These include proteins, amino acids, and organic acids [36]; P. croceum is known to exude

compounds such as oxalic and malic acid [37]. In ectomycorrhizal fungi such as P. croceum, trehalose is the primary storage sugar [38, 39], and this disaccharide may be partially responsible for the selection of specific bacterial communities in mycorrhizospheres [4]. The positive impact of P. croceum on AcH 505 was more significant in microcosms amended with a microbe filtrate. This shows that competition by microbial community may influence the outcome of microbial ICG-001 molecular weight interactions. Schlatter et al. [40] also reported, that the microbial community has an impact: Streptomyces scabiei DL87 promoted Streptomyces lavendulae DL93 in autoclaved, but not in field soil. In general, streptomycetes are competitive because they can derive nutrients from recalcitrant substrates, possess diverse resistance genes and are prolific producers of antagonistic secondary metabolites that inhibit the growth of their competitors [33, 41]. It can also be concluded, that AcH 505

is a competitive streptomycete, as the strain was not affected by the microbe filtrate in the rhizospheres of plants. Fungal responses to soil microbial community and to AcH 505 The soil microbe filtrate inhibited P. croceum, and this inhibition could be due to competition for resources or space, or to antagonism [42]. The first of these possibilities, i.e. competitive inhibition, is perhaps more likely: Schrey et al. [43] obtained evidence that P. croceum many may be particularly tolerant of antagonistic metabolites of Streptomycete isolates from Norway spruce – in an experiment conducted to determine the in vitro activity of Piloderma sp. mycorrhizas against seven fungi, P. croceum was the least severely affected fungus. In this study, Streptomyces affected the growth of Piloderma only under the influence of the microbial filtrate. This indicates that communities of soil microbes carry out a multitude of small-scale processes that can impact bacterium-fungus interactions [1, 36]. Plant rhizosphere reverses the outcome of AcH 505 – P.

Interestingly the less number of gold particles was found in the MSP2 strain as low amount of GS expression and PLG formation. Figure 6 Immunogold localization of PLG in the cell wall of mycobacteria during nitrogen availability. Shown are transmission electron micrographs of the wild type M. bovis and recombinant M. smegmatis strain (MSFP, MSP1 and MSP2) in low and high nitrogen condition. The black arrows shown in the images marked the gold particle around the cell wall periphery in low nitrogen condition. Effect on biofilm formation It was earlier

reported that a ∆glnA1 strain of M. bovis that lack PLG layer in the cell Lorlatinib cost wall was found to be defective in biofilm formation [8]. Our studies on biofilm formation were found to be in accordance with earlier reports. MSFP and M. bovis strains were defective in forming biofilm in high nitrogen on a polystyrene surface. Both strains showed ~ 25% reduction in biofilm formation in high nitrogen condition as compared to low nitrogen condition while M. smegmatis strain showed no difference in the biofilm formation (Figure 7A and B). The pellicle formation for the MSFP and M. bovis strains were also significantly less in high nitrogen as compared to the low

nitrogen condition (Figure 7C). Interestingly, the pellicle formation by M. smegmatis strain complemented with M. bovis glnA1 was enhanced than the wild type. It reiterates the involvement of glnA1 in modulating the cell surface properties of mycobacteria Selleck FK228 [8]. Figure 7 Biofilm and pellicle formation under low and high nitrogen condition. A. M. bovis, wild type M. smegmatis and MSFP were grown 7H9 medium to form biofilm in low and high nitrogen medium. B. Biofilm formation assayed using the 1% crystal violet (CV) staining assay. Cells in low nitrogen (black bars), High nitrogen (crossed bars) and control (grey bars) in 7H9 media were grown in low and high nitrogen on polystyrene plates. The experiments were repeated three times with similar result. Control, medium only. C. Pellicle formation at the air-liquid interface of the standing 7H9 culture by strains M. bovis

(i), M. smegmatis (ii) and MSFP (iii) in low and high nitrogen condition. Results are representative Anacetrapib of at least three independent experiments. LN, low nitrogen; HN, high nitrogen. Discussion Nitrogen metabolism has been studied in detail in industrially important organisms such as Streptomyces and Corynebacteria but there have been very few reports on nitrogen metabolism of mycobacterial species. Earlier, several studies have reported that glnA1 gene is up-regulated in nitrogen starvation in M. tuberculosis and M. smegmatis[5, 12] but this study emphasizes on behaviour of glnA1 locus of M. bovis at both transcriptional and translational levels by altering nitrogen concentration in the medium. Also nitrogen conditions modulate the cell wall properties by altering synthesis of PLG layer in mycobacteria.

Different antibodies have been indifferently used in different studies Deforolimus manufacturer for the detection of the CD133 molecule. In our opinion this can be a highly confusing factor. Indeed, we previously demonstrated, by western blot analysis, that CD133 is expressed at various levels in colon cancers [32, 33] and that different results can be obtained by using different antibodies [34] and similar observations

have been also reported by other Authors [35, 36]. The observation that high CD133 expression has been reported to be a negative prognostic factor for colorectal cancers in several studies using different antibodies strongly suggests an important prognostic significance of its detection [1, 2, 37]. In our study, CD133 also confirmed to be an independent risk factor for a shorter disease-free and overall survival in a multivariate analysis (Tables  4 and 5). These findings are consistent with similar results reported in other human cancers and warrant FK228 order studies on larger cohorts

of patients to further evaluate its suitability as a prognostic marker in the clinical management of colon cancer patients. We observed an unexpected behaviour of CD133 expression which tended to be higher in the lowest grade/stage tumours than in more advanced lesions. Although not expected, this distribution is consistent with previous findings in a mouse model of colon carcinogenesis [38] and in human primary colon cancers [39]. Indeed, in mouse colon Adenosine carcinogenesis we observed a significantly increased expression of CD133, assessed by immunohistochemistry,

in early neoplastic lesions which tended to decrease with tumour development, although remaining always higher in cancer than in normal adjacent tissues [38] and an increased CD133 expression, assessed using a quantitative reverse-transcription PCR, was reported in Dukes A compared to Dukes B and C colon cancers [39]. These findings are in agreement with the proposed ability of the protein to specifically identify tumour initiating cells, important for the growth of both primary and recurrent/metastatic cancers [40] and thus mainly involved in the most active phases of tumour development, i.e., in early lesions (low grade and low stage cancers) as well as in metastatic lesions. Consistent with this hypothesis, CD133 expression has been reported to be highly expressed in colon cancers with early liver metastases and to be a potential biomarker for the early liver metastases [41] and we also previously reported an increased percentage of CD133+ cells, assessed by flow cytometry, in metastatic vs primary colon cancers, [42]. It will be of interest to evaluate the immunohistochemical CD133 expression in the entire process of human colon tumorigenesis (i.e., from early to metastatic lesions) and evaluate how it correlates with tumour development.

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