e., randomized subjects who took any study medication), whether or not they provided any efficacy data. The specific terms used to describe the adverse events in each of the articles were retained during the data extraction. These were then EX 527 cost grouped into relevant categories. Dyspepsia, nausea/vomiting, and abdominal pain were considered separately and also

in aggregate as ‘minor gastrointestinal events’. Dyspepsia was taken to include terms covered by the Medical Dictionary for Regulatory Activities (MedDRA) preferred term ‘dyspepsia’, nonspecific (functional) gastrointestinal disorders, eructation, abdominal/epigastric discomfort, and abdominal tenderness but not abdominal pain. Gastrointestinal bleeding was defined as including all bleeding in the gastrointestinal tract, ranging from a positive stool test to melena. Clinically active gastrointestinal ulcers LCZ696 mouse and perforations were also tabulated, but purely endoscopic findings were not. The term ‘gastrointestinal events’ was

reserved for descriptions of low specificity reported as a sole safety outcome, as well as an overall summary of other events considered in the same publication. Gastrointestinal events that did not match one of these outcome categories were not considered in the analysis (e.g. diarrhea, flatulence, constipation, dry mouth). Data entry was repeated on the 5 % of clinical trial and observational reports that provided the largest number of endpoints. Articles with discrepancies were re-reviewed to reconcile the differences. The risk of experiencing gastrointestinal adverse events after short-term treatment with aspirin was assessed using meta-analytical

methods. We did not include observational studies, as they rarely provided ASK1 detailed data regarding dose and duration of treatment, and they did not directly compare different agents with each other. We OSI-027 solubility dmso included parallel-design, randomized clinical studies with at least one aspirin arm at a dose between 325 and 4,000 mg/day and a treatment duration of at most 10 days. We included only articles that studied aspirin as monotherapy, i.e., not in combination with other active agents (e.g., ephedrine). Vitamin C and caffeine were not considered active components. No exclusions were made with regard to blinding, subject compliance, single vs. multiple dosing, total dosages, or formulations. Crossover trials were excluded because of concerns regarding unknown carryover effects, patient dropout between treatment phases, and within-patient correlations. To avoid including previously reported data, publications describing Bayer-sponsored studies that were included in a previous report [7] were also not included in the current analysis. After these exclusions, a total of 152 studies from 150 publications were considered. In some reports, the number of subjects allocated to each study treatment was stated only as a percentage of an overall total.

The StripAssay was the most analytically sensitive test (Table 2) of those we examined. On the basis of the results obtained with this method in the series of tests conducted with dilution series of mutant KRAS DNA (Figure 6), one could even argue that samples 24 to 30 should be reassigned as mutants (Table 2), thereby changing the false positive rate for the K-ras StripAssay to 0/128 and the false negative rate for TheraScreen DxS to

7/128. However, the interpretation of StripAssay results can be quite problematic Entospletinib cell line for samples whose mutant DNA content is below 1% (see the result obtained with a mutant minority of 0.5% NCI-H620 in Figure 6). Insofar, it was not tested in clinical studies what is a significance of fraction of mutated cells below

1%, selleck kinase inhibitor regardless of the typing method used. During time of submitting this article, company’s software was upgraded to follow more precisely the requirements of ISO15189 norm (scanner calibration standard was added and manual baseline correction feature was removed). It remains to be seen if such changes bring any improvement to diagnostic accuracy. Of the methods examined in this study, the TheraScreen DxS kit was the fastest method and exhibited the highest sensitivity and specificity. However, it was also the most expensive method that is not free of false reactions. Specifically, the kit failed to detect the p.Gly13Cys P5091 mutation in sample 2 because it is not designed to detect this mutation. Although the frequency of the mutations that are not covered by the TheraScreen DxS test is very low and clinically not highly relevant, this nevertheless constitutes an inherent limitation of the kit. In addition, the precise allelic mutation detected by this kit in samples 3, 16, and 18 differed from the consensus result. While this could potentially be due to stochastic variation in the early events of PCR priming, there is no

firm evidence to support this hypothesis. Although discrepancies in the precise check details identity of the mutation are not yet clinically relevant, and these results were not scored as errors in this study, this finding warrants caution when using the ARMS Scorpions assay in different diagnostic setups, where the type of mutation is important (e.g. when looking at the T790M and S768I activating mutations in EGFR genotyping). As discussed above, samples 24 to 30 gave positive results in the StripAssay but were negative when analyzed with the TheraScreen DxS kit, and they seem to have a mutant population in the clinically “grey area,” having less than 1% of the cells in the sample containing KRAS mutation. Ideally, their status should have been resolved by PCR amplicon cloning, followed by sequencing of the clones, digital PCR, or ultradeep sequencing. However, this approach is not practical for routine work and we did not have sufficient DNA to perform this experiment.

Other processes that have been associated with the qT are some slowly relaxing component(s) of qE (Lokstein et al. 1993; Joliot and Finazzi 2010) and light-dependent movements of chloroplasts (Cazzaniga et al. 2013). In practice, there are several arguments making it doubtful that the qT is a reliable measure for state transitions. The slowest relaxation phase, the qI, which may last several find more hours can consist of several processes: photoinhibition of PSII and XC related changes (reviewed by Krause and Jahns 2004) and possibly also state II to state I transitions (Schansker et al. 2006) if a change in the JI amplitude is related to state transitions as suggested by Schreiber et al. (1995) for cyanobacteria.

It should be noted that the rate with which these processes reverse in darkness is not necessarily the same in all photosynthetic organisms. For example, the regeneration of the IP phase parallels the qT phase in pea leaves (Schansker et al. 2006), and it is complete within 15 min, whereas the same process in needles of Pinus halepensis takes 1 h (Schansker et al. 2008). Question 16. Why is far-red light used to determine Trichostatin A mw the F O and F O′ values? For leaves, it is reasonable to assume that under most conditions, nearly all PSII RCs are in the open state (Q A oxidized) following dark adaptation. However, the assumption is not true for heat-stressed

leaves (Ducruet 1999; Tóth et al. 2007b) and leaves that show a high GABA Receptor rate of chlororespiration. Chlororespiration refers to the non-photochemical reduction of the plastoquinone

pool by reducing equivalents derived from Fdred or NADPH in the stroma (Bennoun 2002). Feild et al. (1998) showed a high chlororespiratory activity in light acclimated sunflower leaves following a light-to-dark transition leading to considerably higher F O′ values. This F O′ increase is due to a population of reduced Q A associated with a more reduced PQ pool. There is redox interaction between the PQ-pool and Q A leading to a redox-equilibrium (Diner 1977); for pea leaves, it was shown that a completely reduced PQ-pool (induced by anaerobiosis) is in equilibrium with reduced Q A in 20 % of the PSII RCs (Tóth et al. 2007a). To assure maximum oxidation of the PQ pool, the leaf can be pre-illuminated with FR light. For this purpose, FR light in the 720–735 nm range is normally used. FR light preferentially excites PSI and thereby causes an oxidation of the PQ pool. We note that FR light can induce charge separations in PSII (Pettai et al. 2005; Schansker and check details Strasser 2005). Pettai et al. (2005) demonstrated that FR light at 740 nm still induces a low level of oxygen evolution even though the activity is three times less than that induced by FR light at 720 nm. In practice, FR light induces about 2.5 % of F V associated with Q B − in 50 % of the RCs (Schansker and Strasser 2005).

No difference in virulence was observed between mice receiving tetracycline and control animals. In conjunction, these data suggest that TbrPPX1 may not be an essential gene in bloodstream form T. brucei, neither

in cell culture nor during an in vivo infection. Figure 5 RNAi against TbrPPX1 does not affect proliferation of bloodstream forms in culture. Panel A: Northern blot of two independent bloodstream form clones at 48 h after induction of RNAi. Panel B: determination of generation times in the presence and absence of tetracycline. wt: NYSM parental strain, A3, A5: two independent clones expressing RNAi against TbrPPX1. The figure represents one of two independent experiments. Characterization of recombinant TbrPPX1 TbrPPX1 Selleck RXDX-101 was expressed in E. coli BL21(DE3) cells as a fusion

protein with either an N-terminal GST tag or an N-terminal MBP tag, using the pGST- or pMBP parallel3 vectors [19]. Induction of protein expression with 0.4 mM IPTG overnight at 15°C resulted in mostly soluble fusion protein. The recombinant proteins were isolated by passage over glutathione- or amylose-resin. Both recombinant proteins migrated with the expected molecular masses (TbrPPX1: 42.8 kDa; GST: 26.2 kDa; MBP: 41.2 kDa). Initial activity measurement using pentaAZD5363 sodium triphosphate as AZD6244 cell line a substrate demonstrated that the GST-fusion protein was active, while the MBP fusion construct was completely inactive. In contrast to what was observed with LmjPPX1 [14], recombinant TbrPPX1 was stable after purification, and could be frozen and thawed repeatedly without loss

of activity when kept in elution buffer containing 10% glycerol and 0.5 mM MgCl2. As expected from its sequence, TbrPPX1 proved to be an exopolyphosphatase. Its Km for pentasodium triphosphate as a substrate is 27.2 ± 4.2 μM (n = 3), and its kcat is 8.1 ± 1.5 s-1 (n = 3). Sodium pyrophosphate (Figure 6B) and polyphosphate (average length ~ 17) are neither substrates nor inhibitors. The activity of TbrPPX1 is entirely dependent on divalent cations, and it is not affected by cAMP, deoxynucleoside triphosphates, ATP, sodium pyrophosphate, by basic amino acids that Sirolimus in vivo are enriched in the acidocalcisomes such as arginine, or by long polyanions such as heparin or RNA (Figure 6C). Also, TbrPPX1 is not inhibited by a series of cyclic nucleotide phosphodiesterase inhibitors such as Ro-20-1724, sildenafil, zaprinast, papaverine or etazolate, or the sodium salts of vanadate, fluoride or sulfate. Zn2+ is a strong inhibitor with an IC50 value of 21.3 ± 18.2 μM (n = 3) when the reaction is run in the presence of 1 mM MgCl2 (Figure 6D). Figure 6 Characterization of recombinant TbrPPX1. Panel A: Michaelis-Menten kinetics with pentasodium triphosphate as substrate. Each assay point was done in triplicate (standard deviations are too small to be visible in the graph). A representative graph of three independent experiments is shown. Panel B: Sodium pyrophosphate is neither a substrate for, nor an inhibitor of TbrPPX1.

Prog Polym Sci 2000, 25:1503–1555.CrossRef 8. Van Beilen JB, AZD9291 Poirier Y: Production of renewable polymers from crop plants. Plant J 2008, 54:684–701.PubMedCrossRef 9. Budde CF, Riedel SL, Willis LB, Rha C, Sinskey AJ: Production of poly(3-hydroxybutyrate-

co -3-hydroxyhexanoate) from plant oil by engineered Ralstonia eutropha strains. Appl Environ Microbiol 2011, 77:2847–2854.PubMedCrossRef 10. Fukui T, Suzuki M, Tsuge T, Nakamura S: Microbial synthesis of poly(( R )-3-hydroxybutyrate- co -3-hydroxypropionate) from unrelated carbon sources by engineered Cupriavidus necator . Biomacromolecules 2009, 10:700–706.PubMedCrossRef 11. Kawashima Y, Cheng W, Mifune J, Orita NCT-501 supplier I, Nakamura S, Fukui T: Characterization and functional analyses of R -specific enoyl Coenzyme A hydratases in polyhydroxyalkanoate-producing Ralstonia eutropha . Appl Environ Microbiol 2012, 78:493–502.PubMedCrossRef 12. Matsusaki H, Abe H, Taguchi K, Fukui T, Doi Y: Biosynthesis of poly(3-hydroxybutyrate- co Selleck AR-13324 -3-hydroxyalkanoates) by recombinant bacteria expressing the PHA synthase gene phaC1 from Pseudomonas sp. 61–3. Appl

Microbiol Biotechnol 2000, 53:401–409.PubMedCrossRef 13. Mifune J, Nakamura S, Fukui T: Targeted engineering of Cupriavidus necator chromosome for biosynthesis of poly (3-hydroxybutyrate- co -3-hydroxyhexanoate) from vegetable oil. Can J Chem 2008, 86:621–627.CrossRef 14. Mifune J, Nakamura S, Fukui T: Engineering of pha operon on Cupriavidus necator chromosome for efficient biosynthesis of poly(3-hydroxybutyrate- co -3-hydroxyhexanoate) from vegetable oil. Polym Degrad Stab 2010, 95:1305–1312.CrossRef

15. Tsuge T, Yano K, Imazu S, Numata K, Kikkawa Y, Abe H, Taguchi S, Doi Y: Biosynthesis of polyhydroxyalkanoate (PHA) copolymer from fructose using wild-type and laboratory-evolved PHA synthases. Macromol Biosci 2005, 5:112–117.PubMedCrossRef 16. Pohlmann A, Fricke WF, Reinecke F, Kusian B, Liesegang H, Cramm R, Eitinger T, Ewering C, Pötter M, Schwartz E, Strittmatter A, Voss I, Gottschalk G, Steinbüchel A, Friedrich B, Bowien B: Genome sequence of the bioplastic-producing “Knallgas” bacterium Ralstonia eutropha H16. Nat Biotechnol 2006, 24:1257–1262.PubMedCrossRef 17. Peplinski K, Ehrenreich A, Döring C, Bömeke M, Reinecke F, Hutmacher C, Steinbüchel tuclazepam A: Genome-wide transcriptome analyses of the “Knallgas” bacterium Ralstonia eutropha H16 with regard to polyhydroxyalkanoate metabolism. Microbiology 2010, 156:2136–2152.PubMedCrossRef 18. Brigham CJ, Budde CF, Holder JW, Zeng Q, Mahan AE, Rha C, Sinskey AJ: Elucidation of β-oxidation pathways in Ralstonia eutropha H16 by examination of global gene expression. J Bacteriol 2010, 192:5454–5464.PubMedCrossRef 19. Marioni JC, Mason CE, Mane SM, Stephens M, Gilad Y: RNA-seq: an assessment of technical reproducibility and comparison with gene expression arrays. Genome Res 2008, 18:1509–1517.PubMedCrossRef 20.

Although distribution modeling approaches are available, their applicability for monographic data and for presence-only data in general is often Fosbretabulin ic50 compromised by data scarcity, poor data quality and lack of knowledge of the environmental correlates of species. Our method is precisely targeted at such data and can also be adjusted to accommodate different taxonomical groups by changing the weighting of

interpolation distances for species range generation. Using this new method, we identified and validated centers of Neotropical angiosperm species richness and compared LGX818 manufacturer them to the current protection status and human population density. In addition, we identified areas where insufficient data do not allow for reliable estimates of distribution patterns. This is due to the sensitivity of the distribution

patterns of the narrow endemic species towards sampling effort. In particular, our method might underestimate the numbers and the ranges of narrow endemic species in poorly collected areas. Our maps also indicate areas for further sampling activity, because the available data do not yet allow for robust estimation of species richness patterns. To permit pinpointing of species-rich areas for conservation priorities, a robust estimate of total species richness and narrow endemic species richness selleck chemicals llc is necessary. Therefore, future collection activity should focus on under-sampled areas and under-sampled taxa. Further taxonomic identification of both new and already collected, unidentified specimens is necessary, which requires additional training and support of expert taxonomists. New and reliable data will enable the scientific community to further clarify Neotropical angiosperm distribution and in particular endemism Methocarbamol patterns to improve response to conservation needs. Acknowledgements This study was inspired by the late Wilfried Morawetz, who had the vision of constructing comprehensive species richness maps long before GIS desktops became a standard. Ingo Fetzer, Julio Schneider and two anonymous reviewers greatly helped improve the grammar and readability of the manuscript. CFD acknowledges support by the Helmholtz

Association (VH-NG-247). Open Access This article is distributed under the terms of the Creative Commons Attribution Noncommercial License which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author(s) and source are credited. Appendices Appendix 1 Literature consulted for compilation of the Neotropical angiosperm species distribution database. Anderson C (1982) A monograph of the genus Peixotoa (Malpighiaceae). Contr Univ Michigan Herb 15:1–92 Anderson WR (1982) Notes on Neotropical Malpighiaceae—I. Contr Univ Michigan Herb 15:93–136 Andersson L (1977) The genus Ischnosiphon (Marantaceae). Opera Bot 43:1–114 Andersson L (1985) Revision of Heliconia subgen. Stenochlamys (Musaceae-Heliconioideae).

The Chao model could not be used for the sample PS3 because it did not contain any doubleton sequences. Table 2 Estimations of true diversity of different samples.   Sample Age (d)1 Number of Selleck Thiazovivin sequences Number of OTUs ACE estimate ACE coverage % Chao1 estimate Chao1 coverage % Simpson’s Reciprocal Index Simpson’s Index of Diversity Full-scale process FS1 0 28 23 79.58 28.90 83.17 27.66 2.17 0.54   FS2 1 135 46 97.76 47.06 91.56 50.24 23.55 0.96   FS3 2-3 47 24 103.72 23.14 52.90 45.37 7.40 0.86   FS4 7 50 26 79.66 32.64 66.50 39.10 7.61 0.87   FS5 1 69 37 217.00 17.05 262.00 14.12 5.37

0.81   FS7 0 47 43 252.63 17.02 233.13 18.45 1.45 check details 0.31   FS8 21 118 60 148.23 40.48 160.00 37.50 8.70 0.89   FS9 1 81 33 86.18 38.29 77.10 42.80 14.66 0.93   FS10 2-3 38 31 119.31 25.98 143.67 21.58 2.14 0.53   FS11 12 23 8 12.00 66.67 12.00 66.67 36.14 0.97 Pilot-scale process PS1 4 314

128 672.07 19.05 658.45 19.44 9.26 0.89   PS2 39 163 50 186.78 26.77 179.60 27.84 20.60 0.95   PS3 4 88 10 66.00 15.15 – - 136.71 0.99   PS4 8 60 26 67.45 38.55 66.50 39.10 11.13 0.91   PS5 6 73 25 64.79 38.58 65.50 38.17 16.53 0.94   PS6 10 65 36 104.71 34.38 127.50 50.98 6.69 0.85   PS7 15 78 23 46.36 49.61 65.25 35.25 37.07 0.97   PS8 19 83 28 62.02 45.15 4EGI-1 mouse 76.17 36.76 24.13 0.96 1 Time in days after loading of material into composting unit Discussion The microbial community and its physical and chemical changes during the composting process have received much attention during recent years. However, the picture of the community structure of composting generated by earlier studies,

based on cultivation, Phospholipid Fatty-acid Analysis (PLFA), Denaturing Gradient Gel Electrophoresis (DGGE) or Single Strand Conformation Polymorphism (SSCP), has not been as wide nor as specific at the genus and species level as the one presented here. In earlier studies, such as those by Adams and Frostick [38] and Takaku et al. [39], sequences Celecoxib obtained via DGGE analysis are identified, in some cases to the species level, but the total number of clones sequenced is relatively small. In this study we used a DNA-cloning and sequencing based method to determine, as broadly as possible, the bacterial diversity during the active early phases of composting. The targeted composting units were a pilot-scale unit and a full-scale composting facility. Both units were run semi-continuously using normal source-separated household bio-waste as the substrate. At the full-scale facility also the conditions were realistic with all the challenges of running the unit as efficiently as possible.

Lin AE, Krastel K, Hobb RI, Thompson SA, Cvitkovitch DG, Gaynor EC: Atypical roles for Campylobacter jejuni amino acid ATP binding cassette transporter components PaqP and PaqQ in bacterial stress tolerance and pathogen-host cell dynamics. Infect Immun 2009, 77:4912–4924.PubMedCrossRef 31. Van Deun K, Pasmans F, Ducatelle R, Flahou B, Vissenberg K, Martel A, Van den Broeck APR-246 nmr W, Van Immerseel F, Haesebrouck F: Colonization strategy of Campylobacter jejuni CP673451 manufacturer results in persistent infection of the chicken gut. Vet Microbiol 2008, 130:285–297.PubMedCrossRef 32. Eucker TP, Konkel ME: The cooperative action of bacterial fibronectin-binding

proteins and secreted proteins promote maximal Campylobacter jejuni invasion of host cells by stimulating membrane ruffling. Cell Microbiol 2012, 14:226–238.PubMedCrossRef 33. Bernhardt

TG, de Boer PA: The Escherichia coli amidase AmiC is a periplasmic septal ring component exported via the twin-arginine transport pathway. Mol Microbiol 2003, 48:1171–1182.PubMedCrossRef 34. Kassem II, Zhang Q, Rajashekara G: The twin-arginine translocation system: contributions to the pathobiology of Campylobacter jejuni. Future Microbiol 2011, 6:1315–1327.PubMedCrossRef 35. Frirdich E, Biboy J, Adams C, Lee J, Ellermeier J, Gielda LD, Dirita VJ, Girardin SE, Vollmer W, Gaynor EC: Peptidoglycan-modifying enzyme Pgp1 is required for helical cell shape and pathogenicity traits in Campylobacter jejuni. PLoS Pathog 2012, 8:e1002602.PubMedCrossRef 36. Taveirne ME, Sikes ML, Olson JW: Molybdenum and tungsten in Campylobacter jejuni: their physiological role and identification selleck compound of separate transporters regulated by a single ModE-like protein. Mol Microbiol 2009, 74:758–771.PubMedCrossRef 37. Wilson DL, Bell JA, Young VB, Wilder

SR, Mansfield LS, Linz JE: Variation of the natural transformation Temsirolimus in vivo frequency of Campylobacter jejuni in liquid shake culture. Microbiology 2003, 149:3603–3615.PubMedCrossRef 38. Atack JM, Harvey P, Jones MA, Kelly DJ: The Campylobacter jejuni thiol peroxidases Tpx and Bcp both contribute to aerotolerance and peroxide-mediated stress resistance but have distinct substrate specificities. J Bacteriol 2008, 190:5279–5290.PubMedCrossRef 39. Konkel ME, Kim BJ, Rivera-Amill V, Garvis SG: Bacterial secreted proteins are required for the internaliztion of Campylobacter jejuni into cultured mammalian cells. Mol Microbiol 1999, 32:691–701.PubMedCrossRef 40. Monteville MR, Yoon JE, Konkel ME: Maximal adherence and invasion of INT 407 cells by Campylobacter jejuni requires the CadF outer-membrane protein and microfilament reorganization. Microbiology 2003, 149:153–165.PubMedCrossRef 41. Konkel ME, Hayes SF, Joens LA, Cieplak W Jr: Characteristics of the internalization and intracellular survival of Campylobacter jejuni in human epithelial cell cultures. Microb Pathog 1992, 13:357–370.PubMedCrossRef 42.

Additionally, this file also includes a table about the primers used in this study, a figure reflects the concentration changes of the substrate and of the two intermediates in the course of the PNP degradation and another figure about the specific absorbs curved line which reflects the detected peak by HPLC [13, 21, 22]. (DOC 2 MB) References 1. Bondarenko

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Cloning, expression and purification of recombinant GapA-1 The gapA-1 gene from MC58 was cloned into the expression vector pCRT7/NT-TOPO to facilitate the expression and subsequent purification of 6 × histidine-tagged recombinant GapA-1 (Figure 1a). This was used to generate RαGapA-1. Immunoblot analysis confirmed that RαGapA-1 and anti-pentahistidine antibodies both reacted to the purified recombinant GapA-1 (Figure 1b &1c). Figure 1 SDS-PAGE and immunoblot analysis of

recombinant GapA-1. SDS-PAGE analysis confirms the purity of the recombinant GapA-1 purified under denaturing Kinase Inhibitor Library conditions (a). Immunoblot analysis shows that recombinant GapA-1 is recognized by RαGapA-1 (b) and anti-pentahistidine antibodies (c). Construction of an N. meningitidis gapA-1 null mutant strain To examine the roles of GapA-1 in the meningococcus, a gapA-1 knockout derivative of N. meningitidis MC58 was generated. Immunoblotting using RαGapA-1 showed that GapA-1 could be detected in whole cell lysates of wild-type but not MC58ΔgapA-1 (Figure 2, lanes 1 & 2) confirming that GapA-1 was expressed under the conditions used and that expression had been abolished in the mutant. This analysis further confirmed that the Selleckchem Z-IETD-FMK RαGapA-1 sera did not recognize GapA-2 (37-kDa) under the conditions used. To further confirm that the immuno-reactive protein was GapA-1, a wild-type copy of

gapA-1 was introduced in trans into MC58ΔgapA-1 using plasmid pSAT-14 (Table 1). Introduction of gapA-1

at an ectopic site restored GapA-1 expression (Figure 2, lane 3). Further immunoblot analyses using old a panel of 14 N. meningitidis strains (Additional file 1) including representatives of differing serogroups and MLST-types showed that GapA-1 expression was conserved across all strains (data not shown). Expression was also conserved in N. gonorrhoeae FA1090 (data not shown). These data complement in silico predictions that GapA-1 is universally AZD0156 order present and suggests that GapA-1 is constitutively-expressed across pathogenic Neisseria species. Figure 2 Immunoblot analysis of whole cell proteins from N. meningitidis using RαGapA-1. Analysis of MC58 wild-type, ΔgapA-1 mutant derivative and complemented mutant reveals the absence of GapA-1 in the ΔgapA-1 mutant preparation. Similar analysis shows the abolition of GapA-1 expression in the MC58ΔsiaD ΔgapA-1 mutant compared to the parental MC58ΔsiaD strain. Meningococcal GapA-1 is only surface-accessible to antibodies in the absence of capsule Grifantini et al showed using flow cytometry that GapA-1 was accessible to specific antibodies on the surface of meningococci [27]. However, the methodology used involved pre-treatment of the cells with 70% ethanol to permeabilize the capsule, making it unclear whether GapA-1 was accessible to antibodies in encapsulated bacteria.