It is interesting to note that in this microarray study BBB05 and

It is interesting to note that in this microarray study BBB05 and BBB06 (chbA and chbB, respectively) declined by 40–50% in a rpoN mutant. No changes in BBB04, BBB05, or BBB06 transcription were reported for their rpoS mutant. However, in that study, Fisher et al [18] did not starve cells for GlcNAc, a technique that in our hands results in a modest 2-fold increase in rpoS transcript levels (data not shown), and a corresponding increase in chbC expression (Fig. 3). Additionally,

Lybecker and Samuels [36] CH5183284 recently demonstrated that two rpoS transcripts exist, a shorter RpoN-regulated transcript previously identified by Smith et al. [20] that predominates at high cell density, and a longer transcript that does not possess the canonical RpoN-dependent Ivacaftor nmr promoter whose translation is regulated by the small RNA (sRNA) DsrABb at low cell density. Our physiological and molecular data evaluating chitobiose utilization

(Fig. 4) and chbC expression (Fig. 3) in the wild type versus the rpoS mutant strongly suggests Rabusertib cell line that RpoD and RpoS both regulate chitobiose transport. To determine if the chbC gene has a promoter similar to other RpoS-dependent genes we identified the transcriptional start site (Fig. 6) and the putative chbC promoter (Fig. 7). While not conclusive, it is possible that regulation of chbC by RpoS is through direct binding to the promoter region as the spacing between the -10 and -35 consensus sequences is similar to that of two of the dually transcribed promoters much (Fig. 7). On the other hand, the sequence of the extended -10 chbC promoter element is more like that of the predicted RpoD consensus, and it has been shown that the extended -10 element plays a significant role in sigma factor selectivity in B. burgdorferi [37]. Therefore, it cannot be ruled out that RpoS regulates chbC expression indirectly through an unknown regulator, rather than through direct binding and transcription from the chbC promoter. Conclusion In this study we used a physiologic and molecular approach to demonstrate that chitobiose utilization and chbC expression are dually regulated by RpoD and RpoS. We determined

the chbC transcriptional start site, and identified the putative promoter region. Finally, we provided evidence that the second exponential phase observed in cells cultured in the absence of free GlcNAc is not due to components found in yeastolate, and suggest that the source of GlcNAc in the second exponential phase is sequestered in components of serum and/or neopeptone. Methods Bacterial strains and culture conditions Wild-type B. burgdorferi strain B31-A and rpoS mutant strain A74 were generously provided by Patricia Rosa [38]. All strains were routinely cultured in modified BSK-II medium supplemented with 7% rabbit serum (Invitrogen Corp., Carlsbad, CA) [6]. BSK-II was modified by the replacement of 10× CMRL-1066 with 10× Media 199 (Invitrogen Corp.).

PubMedCrossRef 37 Tao P, Xu DH, Lin SB, Ouyang GL, Chang YD, Che

PubMedCrossRef 37. Tao P, Xu DH, Lin SB, Ouyang GL, Chang YD, Chen Q, Yuan YY, Zhuo XM, Luo QC, Li J, , et al.: Abnormal expression, highly efficient detection and novel truncations of midkine in human tumors, cancers and cell lines. Cancer Letters selleck compound 2007, 253:60–67.PubMedCrossRef 38. Ikematsu S, Nakagawara A, Nakamura Y, Ohira M, Shinjo M, Kishida S, Kadomatsu K: Plasma midkine level is a prognostic factor for human neuroblastoma. Cancer Science 2008, 99:2070–2074.PubMedCrossRef 39. Kang HC, Kim IJ, Park JH, Shin Y, Ku JL, Jung MS, Yoo BC, Kim HK, Park JG: Identification of genes with differential

expression in acquired drug-resistant gastric cancer cells using high-density oligonucleotide microarrays. Clinical Cancer Research 2004, 10:272–284.PubMedCrossRef 40. Thompson DA, Weigel RJ: hAG-2, the human homologue of the Xenopus laevis cement gland gene XAG-2, is coexpressed with estrogen receptor in breast cancer cell lines. Biochemical and Biophysical Research Communications 1998, 251:111–116.PubMedCrossRef 41. Fletcher GC, Patel S, Tyson K, Adam PJ, Schenker M, Loader JA, Daviet L, Legrain P, Parekh R, Harris AL, Terrett JA: hAG-2 and hAG-3, human homologues of genes involved in differentiation,

are associated with oestrogen receptor-positive breast tumors and interact with metastasis gene C4.4a and dystroglycan. NVP-LDE225 nmr British Journal of Cancer 2003, 88:579–585.PubMedCrossRef 42. Liu D, Rudland PS, Sibson DR, Platt-Higgins A, Barraclough R: Human homologue of cement gland protein, a novel metastasis inducer associated with breast carcinomas. Cancer Research 2005, 65:3796–3805.PubMedCrossRef 43. Marquez RT, Baggerly selleck chemicals KA, Patterson AP, Liu JS, Broaddus R, Frumovitz M, Atkinson EN, Smith DI, Hartmann L, Fishman D, et al.: Patterns of gene expression in different histotypes of epithelial ovarian cancer correlate with those in normal fallopian tube, endometrium, and colon. Clinical Cancer Research 2005, 11:6116–6126.PubMedCrossRef 44. Ramachandran V, Arumugam T, Wang HM, Logsdon CD: Anterior gradient

2 is expressed and secreted during the development of pancreatic cancer and promotes cancer cell survival. Cancer Research 2008, 68:7811–7818.PubMedCrossRef 45. Smirnov DA, Zweitzig DR, Foulk 17-DMAG (Alvespimycin) HCl BW, Miller MC, Doyle GV, Pienta KJ, Meropol NJ, Weiner LM, Cohen SJ, Moreno JG, et al.: Global gene expression profiling of circulating tumor cells. Cancer Research 2005, 65:4993–4997.PubMedCrossRef 46. Valladares-Ayerbes M, Diaz-Prado S, Reboredo M, Medina V, Iglesias-Diaz P, Lorenzo-Patino MJ, Campelo RG, Tch MH, Tch IS, Anton-Aparicio LM: Bioinformatics approach to mRNA markers discovery for detection of circulating tumor cells in patients with gastrointestinal cancer. Cancer Detection and Prevention 2008, 32:236–250.PubMedCrossRef Competing interests TAE and DJA are all employees of Healthlinx Ltd, GR is non-executive chairman of Healthlinx Ltd.

aureus has been demonstrated in a number of infection models such

aureus has been demonstrated in a number of infection models such as mastitis [23] and pneumonia [24]. It has also been proposed that α-haemolysin may play a role in colonisation of epithelia by attenuating bacterial clearance from the epithelial surface [25]; this could therefore be of relevance SHP099 nmr to the decontamination of nasal epithelia using PDT. In addition,

α-haemolysin has immunomodulatory properties, notably its ability to trigger the release of pro-inflammatory cytokines such as interleukin-1β [26]; thus inactivation of α-haemolysin by PDT may also protect against harmful inflammatory processes as well as eliminating infecting organisms. The PD0325901 solubility dmso treatment of S. aureus sphingomyelinase with laser light and methylene blue resulted in a significant, dose-dependent reduction in the

enzyme’s activity. Laser light alone also appeared to reduce the activity of sphingomyelinase; however this was found to be not statistically significant. Irradiation of sphingomyelinase with 1.93 J/cm2 laser light in the presence of the highest concentration of methylene blue tested (20 μM) achieved a highly significant reduction in the activity of the enzyme (76%), which was comparable to Doramapimod price the reduction in activity observed for the V8 protease when irradiated for the same time period. This reduction in activity was increased to 92% after irradiation of the enzyme for 5 minutes in the presence of 20 μM methylene blue. Production of sphingomyelinase (β-haemolysin) is thought to be of importance in severe, chronic skin infections, and strains of S. aureus producing high levels of this enzyme have been shown to cause more intense skin lesions than low-producing strains [27]. Inactivation of these toxins may therefore

be of notable relevance to the treatment of superficial staphylococcal skin infections. Sphingomyelinase has recently been shown to kill proliferating T lymphocytes, suggesting a role for this toxin in evasion of the host immune response [28]; hence inactivation of sphingomyelinase by PDT could also reduce the immunomodulatory properties of S. aureus. The photodynamic inactivation of α-haemolysin and sphingomyelinase was shown to be unaffected by the presence of human serum at concentrations resembling the protein content of an acute wound[29], indicating that photodynamic Mannose-binding protein-associated serine protease therapy may be effective in inactivating these virulence factors in vivo. Together with the data showing that PDT using methylene blue and 665 nm laser light is effective against a methicillin-resistant strain of S. aureus, this supports the potential of PDT as a treatment for superficial staphylococcal infections. The precise mechanism of inhibition of these virulence factors has not yet been determined; however it is possible that the reactive oxygen species formed during photosensitisation can oxidise proteins, thereby disrupting their function [13].

8% in our control subjects This frequency is also similar to the

8% in our control subjects. This frequency is also similar to the frequencies NU7441 found in other studies that analyzed GSTP1 polymorphism [18–20]. Some studies have reported a relationship between GST variants and risk of prostate cancer [9, 10, 12, 13, 21]. Investigation of the GSTP1 gene did not reveal any significant association between heterozygous GSTP1 genotype (Ile/Val) and prostate cancer. However, our results suggest that Val/Val genotype of GSTP1

gene could modulate the risk of prostate cancer, even if this association did not reach statistical significance. It should be kept in mind that the inability to reject the null hypothesis could be due to low power of the test because of a relatively Akt inhibitor small sample size. Therefore, the lack of significance does not necessarily mean equality of the distributions. It is plausible that polymorphism at the GSTP1 locus can play an important role in the susceptibility to CUDC-907 different types of cancer. Association of the GSTP1 Val allele with cancer could be expected since the conversion of the amino acid at codon 105 from isoleucine to valine substantially lowers activity of the altered enzyme. It has been predicted

from molecular modelling that the amino acid at this site lies in a hydrophobic binding site for electrophile substrates and thus affects the substrate binding [22]. On the other hand, there are also studies which did not prove any independent effect of this type of polymorphism on the susceptibility for prostate cancer [23–25]. In the present study, we did not observe significantly different crude rates of the GSTM1 and GSTT1 null genotypes in the men diagnosed with prostate cancer and those in the control group. Our

data and the data published by other research groups suggest that differences in the GST frequencies between prostate cancer patients and the control group are relatively small, which therefore makes it difficult to separate the groups from each other new based on statistical data analysis. Once again, the high variability in the groups could mask statistical differences due to low power. The easiest way to improve precision is to increase the number of subjects and patients in the experimental design. However, this may not be applicable to all research conditions due to such factors as additional costs, poorer availability of resources, lower population, which compromises the number of subjects eligible for investigation. In order to achieve a power of at least 80%, we have to identify other explanatory variables and the control for them, and/or apply meta-analysis in order to increase sample size.

The detailed microstructures of the Co3O4 nanosheets were charact

The detailed microstructures of the Co3O4 nanosheets were characterized with TEM. Figure 1b represents typical TEM images of Co3O4 nanosheets. The HRTEM

image shown in the inset of Figure 1b clearly demonstrates lattice fringes with a d-spacing of 0.46 nm (111), matching well with the XRD pattern. To further elucidate Gemcitabine clinical trial the composition, energy-dispersive X-ray spectroscopy was used to determine the nominal stoichiometric atomic ratio of Co and O, as shown in Figure 1c. The chemical composition of the film was investigated by XPS analysis. The spectra (Co 2p and O 1s, as shown in Figure 2) were acquired and processed using standard XPS peak fitting. Two peaks at binding energies of 780 and 795.1 eV were observed from the Co 2p spectra. The tetrahedral Co2+ and octahedral Co3+ contributed to the spin-orbit doublet 2p spectral profile of Co3O4[21]. The relatively sharp peak widths correspond to 2p 1/2 to 2p 3/2 with separation of 15.1 eV, and the weak satellite structure found in the high binding energy side of 2p 3/2 and 2p l/2 transitions

indicate the co-existence of Co(II) and Co(III) on the surface of the material. The Co 2p spectrum is well consistent with the XPS spectrum of Co3O4[22–24]. Figure 2 Co 2 p (a) and O 1 s (b) XPS spectra of Co 3 O 4 sample. The O 1s spectra of the sample was also presented in the inset of the same figure The peak at around 530 eV is due to lattice O, while the peak at about 531.6 eV can be attributed to the low coordinated oxygen ions (chemisorbed oxygen) at the surface [25]. Figure selleck chemicals llc 3a presents the typical current–voltage (I-V) characteristics of RRAM cell with the Au/Co3O4/ITO

structure, measured by sweeping voltage, at a speed of 1 V/s, in the sequence of 0 → 2 → 0 → −2 → 0 V. During the measurements, the bias voltages were applied to the gold top electrode with ITO this website bottom electrode 4��8C as ground. By steady increase of the positive voltages imposed on the RRAM cell, a pronounced change of resistance from the high-resistance state (HRS/OFF) to the low-resistance state (LRS/ON) was observed at about 1.05 V, which is called as the SET’ process, and then the device was set in threshold switching mode (no change in current after this voltage). Figure 3 RS properties of the Au/Co 3 O 4 /ITO memory cells. (a) Typical bipolar resistance switching I-V curves of the Au/Co3O4/ITO cells. (b) Electrical pulse-induced resistance switching of the Au/Co3O4/ITO memory cell at room temperature for 60 s, (inset, data retention of Au/Co3O4/ITO memory cell for >104 s), and (c) I-V curves on log scale. Subsequently, an opposite ‘RESET’ process could also be cited, with the voltage sweep to negative values bringing the device first to an intermediate switching state at −1.53 V that increased up to −1.93 V and, after that, completely to OFF state. The sample exhibits a typical bipolar nature of resistive switching.

From such results, it can be concluded that the positive expressi

From such results, it can be concluded that the positive expressions of CD133 mRNA and CD133 Sotrastaurin price protein positively related to the lymphatic metastasis in GC, which can reasonably be considered as a risk factor to lymphatic metastasis and tumor invasion. Hence, the strategies aimed at the CD133 and SDF-1/CXCR4 modulating axis,

and the molecular pathway for lymphatic metastasis may have important clinical significances to inhibit metastasis of CSCs. Ki-67 is a kind of nuclear protein, which expresses in cellular cycle of G1, S, G2 and M phases, but not in G0 phase. In order to probe the relation of CD133 expression with the proliferation of tumor cells with or without CD133 positivity, the CD133 mRNA expressive level was applied in this study due to the rare CSCs (usually around 1%-5% of total tumor cells) with CD133 protein positivity in tumor as common and the difficulty to identify CSCs as immature tumor cells from matured tumor cells morphologically. From Ruxolitinib solubility dmso current limited information indicated in this investigation of ours, there occurred the significantly higher expression of CD133 mRNA in subgroup with lower Ki-67 LI in comparison with that in subgroup with higher Ki-67 LI. Theoretically, this phenomenon

observed in our study could be elucidated as the various biological profiles in different stage of tumor differential process or in proliferating characterization in the early stage of carcinogenesis and tumor development. And this proliferating characterization would be gradually weakened in tumor development probably. Additionally, in some extent, this higher expression of CD133 mRNA in subgroup with lower Ki-67 LI could also be explained to the resistant potential of CSCs to

anti-cancerous therapy because tumor cells in Phase G0 such as most of CSCs were difficult to be killed by cytotoxin drugs and radiotherapy [18]. On the other hand, for other explanation of this interesting phenomenon with negative relation between CD133 mRNA and Ki-67 LI, as our consideration, it is also contributed to the different proliferating abilities of O-methylated flavonoid matured tumor cells and immature tumor cells of CD133 positivity with some characteristics of CSCs. As well known, CSCs possessed strong RepSox purchase differentiation proficiency, but this proficiency might not mean strong proliferating ability, especially comparing with that of matured tumor cells with CD133 negative expression probably. As there occurred so many kinds of cells in primary lesion and the limitation of only morphological and immunohistochemical observations in this study, the investigation on the both expressions of CD133 and Ki-67 in the same tumor cells should be necessarily considered to carry out in future.

During this period, normal cell division was observed 342 times i

During this period, normal cell division was observed 342 times in the NMFH-1cells, and 70 times in the NMFH-2 cells, and multinucleation was observed 83 times in the NMFH-1cells, and 16 times in the NMFH-2 cells. Regarding normal cell division, which arose in a large number of the traced cells, the constriction proceeded GNS-1480 supplier and the cytoplasm of the parent cell was divided and then the daughter cytoplasm did not fuse from the anaphase to cytokinesis (Figure 4; Additional file 1). Figure 4 Dynamics

of normal cell division by time-lapse video microscopy. The interval of each image is 15 minutes. These images were taken by the incubation imaging system, LCV100, Olympus. The yellow area indicates the location of the nuclei. From anaphase to cytokinesis, the constriction proceeded and the cytoplasm of the parent cell was divided, and then the daughter cytoplasms did not fuse. As for the HER2 inhibitor dynamics of multinucleation, the mononuclear cell moved about extensively, and extended some protrusions. The mononuclear cells were not so much spindle shaped as amoebiform and were round in shape with some protrusions. At the onset of M phase, the nuclear body and the nuclear membrane were disaggregated and could not be seen (prophase), and

then the chromosomes were aggregated and could be seen in the equator YAP-TEAD Inhibitor 1 in vitro of the cell (metaphase). The protrusions receded and the cytoplasm changed spherically, and almost floated. The daughter chromosomes separated (anaphase) and, simultaneously, the cytoplasm developed an elongated shape, the cleavage furrow started to appear, the nuclear membranes emerged, and the cytoplasm started to constrict in the middle (telophase). However, the enough constriction stopped and reversed in the middle of cytokinesis, and the cytoplasm did not divide. As a result, the cell, which included two nuclei, contained one area of cytoplasm (Figure 5; Additional file 2). Similar states were found in the hematoxylin and eosin staining, although each image is presented as a distinct cell (Figure 6). Multinucleation was also observed in a different process between telophase and cytokinesis, such as

before the appearance of the cleavage furrow or at the end of the constriction. Figure 5 Dynamics of multinucleation by time-lapse video microscopy. The interval of each image is 15 minutes. The yellow area indicates the location of the nuclei. In prophase, the nuclear body and the nuclear membrane were disaggregated and could not be seen (a-d), and in metaphase, the chromosomes were aggregated and could be seen in the equator of the cell (e). In anaphase, the daughter chromosomes separated, and in telophase the cytoplasm had an elongated shape, the cleavage furrow started to appear, and the nuclear membranes emerged and the cytoplasm began to constrict in the middle (f). However, the constriction stopped and reversed in the middle of cytokinesis, and the cytoplasm was not divided.

2012;27:783–92 PubMedCrossRef 16 Moldoveanu Z, Wyatt RJ, Lee JY,

2012;27:783–92.PubMedCrossRef 16. Moldoveanu Z, Wyatt RJ, Lee JY, et al. Patients with IgA nephropathy have increased serum galactose-deficient IgA1 levels. Kidney Int. 2007;71:1148–54.PubMedCrossRef 17. Suzuki H, Moldoveanu Z, Hall S, et al. IgA1-secreting cell lines from patients with IgA nephropathy produce aberrantly glycosylated IgA1. J Clin

Invest. 2008;118:629–39.PubMedCentralPubMed 18. Suzuki H, Fan R, Zhang Z, et al. Aberrantly glycosylated IgA1 in IgA nephropathy patients is recognized by IgG antibodies with restricted heterogeneity. J Clin Invest. 2009;119:1668–77.PubMedCentralPubMed CHIR98014 19. Novak J, Julian BA, Tomana M, et al. IgA glycosylation and IgA immune complexes in the pathogenesis of IgA nephropathy. Semin Nephrol. 2008;28:78–87.PubMedCentralPubMedCrossRef 20. Suzuki H,

Kiryluk K, Novak J, et al. The pathophysiology of IgA nephropathy. J Am Soc Nephrol. 2011;22:1795–803.PubMedCrossRef 21. Berthoux F, Suzuki H, Thibaudin L, et al. Autoantibodies targeting galactose-deficient IgA1 associate with progression of IgA nephropathy. J Am Soc Nephrol. 2012;23:1579–87.PubMedCrossRef 22. Ieiri N, Hotta O, Sato T, et al. Significance of the duration of nephropathy for achieving clinical remission in patients with IgA nephropathy treated by tonsillectomy and steroid pulse therapy. Clin Exp Nephrol. 2012;16:122–9.PubMedCrossRef 23. Hotta O, Furuta T, Chiba S, et al. Regression of IgA nephropathy: a repeat biopsy study. Am J Kidney Dis. 2002;39:493–502.PubMedCrossRef 24. Tomana M, Novak J, Julian BA, et al. Circulating immune complexes in IgA nephropathy consist of IgA1 with galactose-deficient hinge region and selleck inhibitor antiglycan antibodies. J Clin Invest. 1999;104:73–81.PubMedCentralPubMedCrossRef

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Chemik 2012, 66:862–867 29 Kutsevol N, Bezugla T, Rawiso M, Bez

Chemik 2012, 66:862–867. 29. Kutsevol N, Bezugla T, Rawiso M, Bezuglyi M, Chumachenko V: In situ synthesis of silver nanoparticles in linear and branched polymer matrices. In International Conference Nanomaterials: Applications and Properties.

Volume 1. Edited by: Pogrebnjak AD. Crimea: Sumy State University Publishing; 2012:1. 30. Zoya Zaheer R: Multi-branched flower-like silver nanoparticles: preparation and characterization. Colloids Surf A: Physicochem Eng Aspect 2011, 384:427–431.CrossRef 31. Chen J, Herricks T, Xia Y: Polyol synthesis of platinum nanostructures: control of morphology through the manipulation of reduction kinetics. Selleckchem Vismodegib Angew Chem Int Ed 2005, 44:2589–2592.CrossRef 32. Herricks T, Chen J, Xia Y: Polyol synthesis of platinum nanoparticles: control of morphology with sodium nitrate. Nano Lett 2004, 4:2367–2371.CrossRef 33. Korichenska O, Kutsevol N, Bezuglyi M: Silver colloid synthesis in linear and branched anionic polymer matrices by using ascorbic acid as reductant. Int Conf Nanomaterials Appl Prop 2013, 2:171–173. Competing interests The authors declare that they have no competing GSK872 ic50 interests.

Authors’ contributions VC and NK carried out the polymer and nanoparticle synthesis, polymer https://www.selleckchem.com/products/torin-1.html characterization, plasmon absorption study, and statistical analysis. MR carried out the SEC measurements and participated in the design of study and coordination. MS and CB carried out the TEM experiment. All authors read and approved the final manuscript.”
“Background Tissue engineering (TE) is the discipline which includes both creation of the new tissue and design and realization of the cells on substrates [1, 2]. Substrates STK38 play a key role in creation of the cell environment [3]. To guide the organization, growth, and differentiation of cells in TE constructs, the biomaterial scaffold should be able to provide not only a physical support but also the chemical and biological clues needed in forming functional

tissue [4–6]. Biomaterials and various synthetic and natural materials, such as polymers, ceramics, metals, or their composites, have been investigated and used in different manners [5, 7]. Polymeric materials have been widely studied as substrates for tissue engineering due to their unique features such as mechanical properties, high availability, low cost, and relatively easy design and production [6, 8]. However, only a few polymers provide the biocompatibility needed to be used with the cells in vitro and in vivo[9]. High-density polyethylene (HDPE) has been extensively used for application such as the part of orthopedic implants [10]. To induce a regeneration process and to avoid the problems due to tissue replacement with a permanent implant, research has been oriented towards the development of polymers that would degrade and could be replaced by human tissue produced by the cells surrounding the material [9].

Current Genetics 2004, 45:214–224 CrossRefPubMed Authors’ contrib

Current Genetics 2004, 45:214–224.CrossRefPubMed Authors’ contributions AS performed microarray analysis, constructed mutant strains, did PCR analysis and contributed to analysis of array data. TA cultured and characterized biofilms, and collected and purified RNA for array analysis. KM contributed to analysis of array data, particularly to K means analysis. SB performed TEM analysis. AN was primarily responsible for the design and analysis of the Thiazovivin manufacturer microarray experiments and especially the comparison with other data sets. PAS performed SEM and microscopy, contributed to array analysis and was primarily responsible for biofilm experimental design.”
“Background Pseudomonas aeruginosa

is an opportunistic, non-fermentative, gram-negative rod which is an important cause of nosocomial infection leading to septicemia and death [1]. The mortality rate is higher than bacteremias caused by other gram-negative opportunistic pathogens. One of the most important features of the bacterium is its resistance to various antibacterial agents [2,3], and even newly developed antibiotics have failed to reduce the mortality rate associated with this organism Selleckchem Pinometostat [4]. There is increasing interest in bacterial virulence factors

as a basis for effective vaccines and immunotherapies. Several extracellular products fromP. aeruginosa such as exotoxin A, exoenzyme S, MLN2238 phospholipase and hemolysins have been studies as potential virulence factors [5]. The role of exotoxin A

in the mortality of experimentally-infected animals has been demonstrated [6] and the LD50 of the exotoxin reported to be 60–80 ng/mouse [7]. Following a single injection of 80 ng of exotoxin A, necrosis, and Terminal deoxynucleotidyl transferase cellular swelling were detected in liver within 48 h [7]. Hemorrhage in the lungs and necrosis in the kidneys were also reported [7,8]. In eukaryotic cells, when exotoxin A turns into an activated enzyme, transfer of an adenosine diphosphate ribose moiety from NAD led to inactivation of elongation factor 2 and inhibition of protein synthesis [7]. Furthermore, the pre-existence of a high titer of anti-exotoxin A antibody reportedly increased the survival rate in patients withP. aeruginosa bacteremia [9]. This study was performed to determine the immunogenicity of a toxoid produced from exotoxin A ofP. aeruginosa in a mouse burn model. Methods Preparation of exotoxin A A toxigenic strain ofP. aeruginosa (PA 103) was used for exotoxin A preparation. Exotoxin A was partially purified according to the method described by Pollack et al. [10] and Homma et al. [11].P. aeruginosa was inoculated into tryptic soy agar and incubated at 37°C for 24 h in ambient conditions. The growth product of the slant cultures was inoculated into 500 mL of Muller-Hinton broth and incubated at 37°C for another 24 h in ambient conditions.