Thus, the best results were obtained when the final concentration of the three primer sets, MgCl2, and Taq polymerase was increased respectively to 0.8 μM, 3 mM and to 1.5 U and the m-PCR was carried

out in a final volume of 50 μl. The thermal cycler parameters of the m-PCR were similar to those of the individual PCR using 61°C as an optimal annealing temperature. Positive and negative control DNA samples were run in each experiment. PCR products were analyzed in 1.2% agarose gel electrophoresis, stained with ethidium bromide and visualised with ultraviolet transillumination. All PCR reactions assessing limits of detection or specifiCity were performed in duplicate. Sensitivity and specifiCity of the m-PCR Sensitivity of the PCR assay was checked using serial fold dilutions of bacterial suspension AZD2281 ic50 of references strains AB7, iB1 and Nine-Miles at 107 bacteria per ml. Simulated positive samples were also obtained by adding

50 μl of bacterial suspension dilution to 50 μl of bacteria-free vaginal swab extract or milk sample. These preparations were then submitted to extraction procedures and to simplex and m-PCR as described above. The specifiCity of the PCR was assessed on 20 strains of Cp. abortus, 5 strains of Cp. pecorum and, 4 strains of C. burnetii Adriamycin from our laboratory bacteria collection and on some isolates suspected to be present into tested clinical samples: Brucella melitensis, Brucella abortus, Brucella suis, Escherichia coli, Bacillus cereus, Listeria monocytogenese, Salmonella abortus ovis, Salmonella Typhimurium, Staphylococcus aureus, Staphylococcus chromogenese, Staphylococcus hominis, Streptococcus dysgalactiae and Streptococcus ogalactiae, Mycobacterium avium, Legionella pneumophila. In addition, RFLP-PCR analysis was carried

out as a confirmatory test for the PCR reaction specifiCity. Thus, 10 μl of amplification products obtained from naturally infected clinical samples and those obtained from 102 genomic DNA templates of the reference strains AB7, IB 1, Nine Miles were subjected to 5 units http://www.selleck.co.jp/products/Abiraterone.html of AluI restriction enzyme (Promega, Charbonnières-Les-Bains, France) in a 20 μl final volume for 3 hours at 37°C. The digested products were examined by using 2% agarose gel stained with ethidium bromide and viewed under UV illumination. In addition, PCR products amplified from clinical samples were purified with a QIAquick PCR purification Kit (Qiagen, Courtaboeuf, France) and directly sequenced with an ABI PRISM 310 genetic analyzer (Applied Biosystems). Isolation of Chlamydophila and Coxiella strains Pathogen isolation was performed to confirm the presence of the involved bacteria, on 20-different PCR positive samples showing high ethidium bromide intensity on agarose gel. Chlamydophila strains isolation were performed using both plaque assays and blind passages on McCoy monolayer cell cultures [27].

For example, transmembrane

proteins involved in the transport of metallic ions appear to play an important role in microbial pathogenesis [51] as demonstrated in the Cu2+-ATPase mutants of learn more Listeria monocytogenes [52] and Criptococcus neoformans [53] that show reduced virulence. In the latter case, the Δvph1 mutant did not display laccase activity, which is an essential virulence factor of this pathogen [53]. Moreover, an ATP-binding cassette (ABC) transporter listed in Table 2 is overexpressed in mycelia cultured in keratin, suggesting its involvement in T. rubrum pathogenicity. In addition, the strain carrying a disrupted version of this MDR gene (ΔTruMDR2) showed low infectious capability characterized by reduced growth of T. rubrum on human nails [40]. Conclusions We identified 575 novel ESTs and obtained new molecular data related to T. rubrum growth, pH and carbon source signaling, and stress responses to antifungal challenges. It is clear that additional studies are necessary to define the functioning of whole genes and fully understand the regulation of these complex adaptive responses. However, the various ESTs identified in this work provide new insights into different aspects of T. rubrum biology,

revealing new sources for functional genome analysis. T. rubrum genes that encode putative proteins similar BVD-523 mouse to virulence factors described for other fungi were among the ESTs identified. The transcriptional profile also suggested that several genes could function in environmental

stress responses. Thus, our study can help to better understand the molecular mechanisms of the adaptive responses possibly involved in dermatophyte infection and antifungal resistance. Methods Strains and culture conditions The H6 (ATCC MYA-3108) and F6 mutant (a fluconazole-resistant strain isolated in our laboratory) strains of T. rubrum were cultured on Sabouraud dextrose agar plates (SDA) as described earlier [54]. Phosphoprotein phosphatase The F6 cultures were supplemented with fluconazole (200 μg/mL). Conidia from these strains were used to construct the cDNA (library 1) or were inoculated in Sabouraud dextrose broth (SDB) and incubated for 72 h at 28°C on an orbital shaker at 180 rpm. The resulting mycelia were aseptically transferred to the desired culture media, and these were used to construct each of the SSH libraries. Construction of the libraries One cDNA library (Library 1) and nine SSH libraries (Libraries 2 to 10) were constructed. The SSH libraries were performed between the tester and driver DNA, with the cDNA population containing the differentially expressed transcripts being the tester, and the reference cDNA (control) being the driver.

Restriction enzymes and T4 DNA ligase were purchased from Roche Applied Science or New England Biolabs and used according to the manufacturer’s instructions. PCRs were performed using either Goldstar Red Taq polymerase (Eurogentec) or iProof High-Fidelity DNA polymerase (Bio-Rad) according to the manufacturer’s instructions. Nucleotide sequencing was performed using the ABI Prism BigDye Terminator Ready Reaction cycle sequencing kit, version 3.1 (Perkin Elmer-ABI). Nucleotide sequences were analyzed by using the CloneManager and Phred/Phrap/Consed software. Identification

of transcription start site The start point of cpoA transcription was determined by rapid amplification of cDNA ends (5′ RACE) as described learn more previously [49] using RNA of S. pneumoniae R6 isolated at a culture density of 40 NU. The primer

cpoARACE2 was used for reverse transcription of RNA ligated to the RNA adapter, and the nested primer and cpoARACE1 was used for amplification of cDNA (for primers, see Additional file 2: Table S1 and S2). Construction of EPZ004777 delivery cassettes, plasmids and mutants To identify the initiation site of cpoA translation, fusions of two DNA fragments with the lacZ reporter gene were constructed. They contained P cpoA (i) together either with two potential start codons (ATG1 and ATG2 in Figure 1B), (ii) with a mutation in ATG2 (ATA), or (iii) with ATG1 only. The three fragments were amplified from chromosomal

DNA of S. Amrubicin pneumoniae R6 by using the primer pairs PcpoA_Eco_f/PcpoA_r2, PcpoA_Eco_f/PcpoABam_r1a and PcpoA_Eco_f/PcpoABam_r1b, cleaved with EcoRI and BamHI, and ligated with the EcoRI/BamHI-digested translation probe vector pTP2. The desired plasmids, pTP2PcpoA-ATG21, pTP2PcpoA-ATG1a and pTP2PcpoA-ATG1b were isolated after transformation of E. coli DH5α and subsequently used to transform S. pneumoniae R6; alternatively plasmids were directly transformed into S. pneumoniae R6. DNA from TetR transformants was PCR-amplified and sequenced to confirm the presence of the lacZ fusions in the resulting strains R6-PcpoA-ATG21, R6-PcpoA-ATG1a and R6-PcpoA-ATG1b. In-frame deletions in cpoA, spr0982, spr0983, obg, or spr0985 were constructed via a two-step process in which the central part of the respective gene(s) was first replaced with the Janus cassette [50] that confers a KanR StrS phenotype in a StrR background. In the second step, the Janus cassette was deleted, thus restoring the original StrR phenotype. The constituents of ‘replacement fragments’ and ‘deletion fragments’ used in the first and second steps of each deletion were amplified from chromosomal DNA of S. pneumoniae R6 by using the primer pairs listed in Additional file 2: Table S2. To generate a ‘replacement fragment’, two PCR products of 0.

The 243 individuals experienced a total of 266 clinical malaria attacks (mean = 1.09, 95%CI: 0.88-1.30). The number of clinical malaria attacks experienced per individual varied from 0 (140 individuals) to 7 (1 individual). Recordings of the entomological inoculation rate indicated a mean of 170 infected bites/person during this time period. Twenty-nine percent of the seronegative individuals (with VS-4718 ic50 no detected anti-MSP1 block2 antibodies) experienced a clinical attack during that period, compared

with 15% of individuals with anti-block2 antibodies. Using a Poisson regression model, the crude estimates of the Incidence Rate Ratio (IRR) of malaria attacks associated with the presence of antibodies to one allelic family AUY-922 solubility dmso or ≥ 2 families (no antibodies as reference group) were 0.55 (95%CI: 0.38-0.80) and 0.21 (95%CI: 0.08-0.58),

respectively (P < 0.0001). In a multivariate Poisson regression analysis, this association was independent of haemoglobin type or ethnic group. However, it was confounded by age, i.e. within the age groups, there was no significant association between the incidence of clinical malaria attacks and the number of MSP1 block2 allelic families recognized. Analysis of the response during a high transmission season To study the impact of novel infections during the transmission season on the humoral response to MSP1 block2, we investigated the fingerprick blood samples collected from 25 seropositive individuals throughout the high transmission season. By the end of December 1998, namely five months after the cross-sectional sampling, the anti-MSP1 block2 antibody level was reduced by ≥ 2-fold in 15 subjects (59%), had varied less than 2-fold in 9 individuals (36%) (typical profiles are shown in Figure 8 upper and middle panel, respectively) and was ≥ 2-fold higher in one

individual (Figure 8, lower panel). Importantly, when a Phosphoglycerate kinase change was observed, it concerned the intensity of the reaction but not its specificity. In other words, responding individuals usually reacted with the same pool(s) and within the pool(s) with the same individual peptide(s) before and after the transmission season. In none of the studied individuals were novel antibody specificities stably acquired during that time period, despite an elevated infection rate. Figure 8 Typical profiles of the temporal evolution of MSP1 block2- specific IgG before and after the 1998 rainy season. Antibodies were assayed from 25 individuals in August 1998 (yellow) and December 1998, i.e. after a rainy season when each inhabitant was exposed to a mean of 170 infected bites. Anti-MSP1 block2 specific IgG was assessed by ELISA on 16 pools of biotinylated peptides.

“
“Background Dye-sensitized solar cells (DSSCs) are attracting attention globally because of their

low cost, high energy conversion efficiency and potential applications [1–4]. Graphene has been extensively utilized in organic photovoltaic (PV) cells owing to its excellent optical and electrical characteristics, which are exploited in transparent conductive films or electrodes [5–8]. Some researchers have reported on composite graphene-TiO2 photoelectrodes in DSSCs [9–12]. Fang et al. [9, 10] discussed the effect of the amount of graphene on the structures and properties of DSSCs. DSSCs with the optimal composite TiO2 film can achieve a photoelectrical conversion efficiency of 7.02%. Graphene is also commonly find more used in graphene-based counter electrodes in DSSCs [13–15]. The conventional counter electrode is platinum (Pt) because of its outstanding conductivity, catalytic activity, and stability when in contact with an iodine-based electrolyte. The expensive Pt can be replaced with graphene films in DSSCs without significantly sacrificing photoelectrical efficiency.

This replacement can simply reduce the cost of the fabrication process [13]. Zhang et al. [14] grew DSSCs with graphene-based counter electrodes, which exhibited a photoelectrical conversion efficiency of as high as 6.81%. Double-layer photoelectrodes have been used to increase the photoelectrical conversion efficiency of DSSCs. Many investigations have focused on modifying the nanostructures of TiO2 photoelectrodes NVP-BGJ398 cell line to nanospheres, nanospindles, nanorods, nanowires, and others [16–20]. Many special nanostructures of photoelectrodes can increase Thymidylate synthase the scattering of light and improve the performance of DSSCs [16, 17]. This work develops a new TiO2/graphene/TiO2 sandwich structure for photoelectrodes. A thin layer of graphene was inserted into the traditional TiO2 photoelectrode layer, making it a double layer. DSSCs with the traditional structure were also fabricated and the characteristics

of the prepared DSSCs were compared. The DSSC with the TiO2/graphene/TiO2 sandwich structure exhibited excellent performance and higher photoelectrical conversion efficiency. This improvement is associated with the increase in electron transport efficiency and the absorption of light in the visible range. Methods Preparation of TiO2 photoelectrodes The TiO2 slurry was prepared by mixing 6 g of nanocrystalline powder (P25 titanium oxide; Evonik Degussa Japan Co., Ltd., Tokyo, Japan), 0.1 mL Triton X-100, and 0.2 mL acetylacetone. The slurry was then stirred for 24 h before being spin-coated on ITO glass substrate at a rotation rate of 2,000 or 4,000 rpm. Following the deposition of graphene, the above procedure was carried out in the fabrication of DSSCs with the TiO2/graphene/TiO2 sandwich structure. The as-prepared TiO2 photoelectrodes were dried and annealed at 450°C for 30 min.

Control, NC and CXCR7shRNA transfected cells adhered equally to BSA-coated dishes. Together, these results indicate that treatment with CXCL12 increases adhesive ability of SMMC-7721 cells and CXCR7 silencing results in decreased adhesive ability. Figure 5 Effect of CXCR7 silencing on HCC cells adhesion in vitro. SMMC-7721 cells were treated as described in Materials

and Methods. SMMC-7721 cells displayed an enhanced cell adhesion to LN or FN in the presence of CXCL12. Cells transfected with CXCR7shRNA showed significantly reduced ability of adhesion to LN or FN compared with control and NC cells. Each bar represents mean ± SD from three independent experiments. *p < 0.05 (as compared with untransfected cells). CXCR7 silencing inhibits tumor cell-induced tube formation in vitro To address whether CXCL12/CXCR7 interaction could mediate in vitro tumor Selleckchem LY3023414 cell-induced tube formation, a coculture system was used in which HUVECs were induced by HCC cells to form capillary-like structures. The tube formation of HUVECs on the Matrigel was quantified by measuring the tube number. As shown in Fig. 6, control and NC cells induced HUVECs to differentiate into capillary-like structures within 24 h. In contrast, SMMC-7721 cells transfected with CXCR7shRNA markedly inhibited tumor cell-induced BMN 673 research buy tube formation. HUVECs showed a significant 32% decrease in the number

of tubes after transfecting SMMC-7721 with CXCR7shRNA. Figure 6 Effect of CXCR7 silencing on tube formation induced by SMMC-7721 cells. HUVECs were cocultured with SMMC-7721 cells, as described

in Materials and Methods. Inhibition of CXCR7 expression in SMMC-7721 cells impaired tube formation induced by SMMC-7721 cells. Each bar represents mean ± SD from three independent experiments. *p < 0.05 (as compared with control cells). CXCL12 induces VEGF secretion through CXCR7 in HCC cells To evaluate whether CXCL12 contributes to proangiogenic factor secretion in HCC cells, we treated SMMC-7721 cells with CXCL12 (100 ng/ml) and measured secretion of proangiogenic factor VEGF by ELISA analysis. As shown in Fig. 7, VEGF secretion increased significantly when SMMC-7721 cells were treated with CXCL12 for 24 h. To further investigate Interleukin-2 receptor whether VEGF secretion was mediated by CXCR7, CXCR7 expression was inhibited by RNA interference before treatment with CXCL12. Significant reduction in VEGF secretion was observed in CXCR7shRNA cells compared with control and NC cells. Thus, these findings indicate that CXCL12 can induce VEGF secretion in SMMC-7721 cells and that CXCR7 can serve as a factor involved in regulation of secretion of VEGF. Figure 7 CXCL12 induces VEGF secretion through CXCR7 in SMMC-7721 cells. SMMC-7721 cells were plated in the 24-well plates. SMMC-7721 cells were serum starved for 24 h, and the cells were treated with CXCL12 (100 ng/ml).

Coverage The coverage of reads mapped to a reference genome was assessed using BEDTools ( https://​github.​com/​arq5x/​bedtools2) and the genomeCoverageBed function. Plasmid analysis A query sequence

of 9299 bases, positions 3036 to 12334 from Lens plasmid pLPL (Accession: NC_006366) was used to search blast databases using blastall (blastn program) from NCBI. Overview of genome similarity BRIG (BLAST Ring Image Generator) was used to produce an image to illustrate the similarity between the Corby genome and one sequence from each of the BAPS clusters (except for Clusters 1 and 2 where two sequences were included, one from each clade on the phylogenetic tree produced from SBT data). Similarity was determined using BLASTn. Gene content analysis A novel method was used to cluster the genes from ICG-001 all the genomes in the study. This method we have termed CoreAccess is reported in full in a paper currently under preparation. Briefly, the protein sequences of all genes from the genomes were

used as input for the program cd-hit [49]. These genes were either those already annotated in the sequence files of the GenBank genomes or those predicted using Glimmer3 [50] trained using the Corby sequence genes. The proteins were clustered using cd-hit using a hierarchical approach, first clustering at a high percentage cut-off and then stepwise lowering of the cut-off and clustering the clusters from the previous step. The final cut-off was 80%. This hierarchical approach overcomes errors that can arise in single R788 ic50 step clustering as described on the cd-hit website (cd-hit.org). The hypothesis underlying this methodology is that the clusters contain homologous proteins from the different genomes and as such represent groups of proteins with the same or similar function from the different genomes. In order to be able to search the clusters and find for example genes shared by all the genomes, the information about the clusters

in the cd-hit output was collated into a sqlite3 database using tools within the Core Access suite. Phylogenetic Tree construction second Maximum likelihood tree phylogenetic trees were produced from mutiple fasta files by the MEGA software package [51] using the Tamura-Nei model, and testing the phylogeny with 500 bootstrap replicates. To construct a tree from the gene content analysis, the database generated by CoreAccess was queried using SQL so that the presence/absence of a protein representative from each strain in every cluster was recorded to produce a phylip compatible discrete state (binary 0/1) character matrix. The seqboot program for the Phylip package [52] was used to create 100 bootstrap replicates using the Discrete Morphology data type and Non-interleaved as parameters.

The stromatolites can be classified as close laterally linked hemispheroid (LLH-C) type. Maximum and minimum thickness of laminaes is between 0.55 and 4.93 mm, respectively. Laminaes are wavy in nature, show low synoptic relief

and high inheritance. In profile section, the laminaes are gently convex. This finding has a tremendous bearing on the evolution of hitherto unknown early life forms in the Archean Bundelkhand craton vis-à-vis central Indian shield. find more Pati, J. K. (2005). The Dhala Structure, Bundelkhand craton, Central India—a new large Paleoproterozoic impact structure (abstract), Meteoritics & Planetary Science 40 (Supplement): A121. Pati, J. K., Reimold, W.U., Koeberl, C. and Pati, P. (in press).

The Dhala Structure, Bundelkhand Craton, Central India—eroded remnant of a large Paleoproterozoic impact structure. To appear in the Meteoritics & Planetary Science. Schopf, J. W., Kudryavtsev, A. B., Czaja, A. D., Tripathi, A. B. (2007). Evidence of Archean life: Stromatolites and microfossils. Precambrian Research, 158:141–155 E-mail: jkpati@yahoo.​co.​in The Minimal Size of Cells: An Experimental Approach Based on Liposomes Tereza Pereira de Souza1, Pasquale Stano1, Pier Luigi Luisi1 Biology Department, University of RomaTre, Viale G. Marconi 446; 00146 Rome, Italy In the last few years the notion of the “minimal Afatinib purchase cell”, as a form of minimal life, has gained considerable attention both from the theoretical and experimental Adenosine point of views (Luisi, 2006; Luisi et al. 2006). This concept is important for assessing the minimal and sufficient conditions for cellular life, and also to gain an insight of the early cells, conceivably much simpler than the modern cells. There are two sides to the notion of minimal cell: one side is the question of the minimal genome, namely the minimal number of expressed genes that permit the functioning of the cell (usually seen in terms of the triad self-maintenance, reproduction,

and evolvability). The other side to it concerns the minimal physical dimension of the cell the question, namely, on the dimension that still permits a cellular life. These two aspects minimal genome and minimal size are obviously connected to one another, being also related to evolutionary paths and to the environment composition. Here we propose to examine the question of the minimal physical size of cells by using liposomes with entrapped the complete ribosomal machinery for protein expression (enhanced green fluorescence protein, EGFP). Liposomes are formed by film or ethanol injection method. The synthesis outside vesicles was inhibited using the EDTA, RNAse or protease, with the inhibitor being added just after vesicles formation. The system with the addition of inhibitor inside and outside of vesicles formed our negative control.

pylori, an organism that has impacted more than half of the world’s population and continues to pose great risk to human health because of its association with gastric cancer and MALT lymphoma. Genetic heterogeneity of the bacterium within a host population as shown in this study should be taken into account when studying the epidemiology

and pathogenesis of H. pylori since there is clearly variation in incidence and severity of the disease in different populations. Methods Source of gastric biopsies and culture of H. pylori isolates Gastric biopsies were collected as part of a large-scale gastric cancer study conducted in symptomatic patients APO866 undergoing gastroenterological examination at the Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia. All biopsies were obtained with the informed consent of the patients and this study was approved by the Human Ethics Committees of the University of New South Wales and the University of Malaya. Based on endoscopic and histological examinations, patients were diagnosed as having

gastric cancer or functional dyspepsia. All except seven samples were from patients with functional dyspepsia as shown in Table 2. H. pylori was cultured by inoculating biopsies on Campylobacter selective agar (CSA) containing 4% blood base agar No. 2 Veliparib (Oxoid), defibrinated horse blood (Oxoid), and one vial of Skirrow’s supplement (Oxoid) containing 2.5 mg Trimethoprim, 5.0 mg Vancomycin, and 1250 IU polymyxin B. Primary cultures were incubated at 37°C with 10% CO2 in a CO2 incubator (Plymouth, USA) for up to 10 days, observing daily for growth. For isolation of pure cultures a single colony was picked and subcultured onto CSA for four days. Identification of H. pylori was based on microscopic morphology and biochemical testing (urease, oxidase and catalase). One isolate from

each biopsy was selected for this study and 78 isolates were obtained from patients of different ethnic background, including 27 Chinese, 35 Indian and 16 Malay (Table Orotic acid 2). We used all Malay biopsy samples available. Despite the fact that this study spanned a period of four years the number of Malay subjects from whom H. pylori could be cultured was low which reflects the relative low prevalence in this population. Isolates from this study are available to researchers upon request to HM. Chromosomal DNA purification One plateful of bacterial culture was collected and suspended into 215 μl of Tris (50 mM), 15 μl of EDTA (0.5 M) and incubated for 10 min. Two μl of proteinase K (10 mg/ml) and 20 μl of SDS (10%) were added followed by incubation at 50°C for a minimum of 2 h or until clear. One μl of RNase (10 mg/ml) was added and incubated at 65°C for an additional 20 min. the mixture was then transferred into a 1.

Gruening P, Fulde M, Valentin-Weigand P, Goethe R: Structure, regulation, and putative function of the arginine deiminase system of Streptococcus suis. J Bacteriol

2006, 188:361–369.CrossRefPubMed 14. Winterhoff N, Goethe R, Gruening P, Rohde M, Kalisz H, Smith HE, Valentin-Weigand P: Identification and characterization of two temperature-induced surface-associated proteins of Streptococcus suis with high homologies to members of the Arginine Deiminase system of Streptococcus pyogenes. J Bacteriol 2002, 184:6768–6776.CrossRefPubMed 15. Handfield M, Brady LJ, Progulske-Fox A, Hillman JD: IVIAT: a novel method to identify microbial genes expressed specifically during human infections. Trends Microbiol 2000, 8:336–339.CrossRefPubMed 16. Rollins SM, Peppercorn A, Hang L, Hillman JD, Calderwood SB, Handfield M, Ryan ET: In vivo induced antigen technology (IVIAT). Cell Microbiol

2005, 7:1–9.CrossRefPubMed www.selleckchem.com/products/azd4547.html 17. Salim KY, Cvitkovitch DG, Chang P, Bast DJ, Handfield M, Hillman JD, de Azavedo JC: Identification of group A Streptococcus antigenic determinants upregulated in vivo. Infect Immun 2005, 73:6026–6038.CrossRefPubMed 18. John M, Kudva IT, Griffin RW, Dodson AW, McManus B, Krastins B, Sarracino D, Progulske-Fox A, Hillman JD, Handfield M, Tarr PI, Calderwood SB: Use of in vivo-induced antigen technology for identification of Escherichia coli O157:H7 proteins expressed during human infection. Infect Immun 2005, 73:2665–2679.CrossRefPubMed 19. Harris JB, Baresch-Bernal A, Rollins

SM, Alam A, LaRocque RC, Bikowski M, this website Peppercorn AF, Handfield M, Hillman JD, Qadri F, Calderwood SB, Hohmann E, Breiman RF, Brooks WA, Ryan ET: Identification of in vivo-induced bacterial protein antigens during human infection with Salmonella enterica serovar Typhi. Infect Immun 2006, 74:5161–5168.CrossRefPubMed 20. Hang L, John M, Asaduzzaman M, Bridges EA, Vanderspurt C, Kirn TJ, Taylor RK, Hillman JD, Progulske-Fox A, Handfield Galeterone M, Ryan ET, Calderwood SB: Use of in vivo-induced antigen technology (IVIAT) to identify genes uniquely expressed during human infection with Vibrio cholerae. Proc Natl Acad Sci USA 2003, 100:8508–8513.CrossRefPubMed 21. Bethe G, Nau R, Wellmer A, Hakenbeck R, Reinert RR, Heinz HP, Zysk G: The cell wall-associated serine protease PrtA: a highly conserved virulence factor of Streptococcus pneumoniae. FEMS Microbiol Lett 2001, 205:99–104.CrossRefPubMed 22. Chen C, Tang J, Dong W, Wang C, Feng Y, Wang J, Zheng F, Pan X, Liu D, Li M, Song Y, Zhu X, Sun H, Feng T, Guo Z, Ju A, Ge J, Dong Y, Sun W, Jiang Y, Wang J, Yan J, Yang H, Wang X, Gao GF, Yang R, Wang J, Yu J: A glimpse of streptococcal toxic shock syndrome from comparative genomics of S. suis 2 Chinese isolates. PLoS ONE 2007, 2:e315.CrossRefPubMed 23. Berry AM, Lock RA, Hansman D, Paton JC: Contribution of autolysin to virulence of Streptococcus pneumoniae. Infect Immun 1989, 57:2324–2330.PubMed 24.