Microbial Pathogenesis

The transcriptome of Salmonella typhimurium 14028S wild-type and soxs mutant in N-minimal medium.

The Transcriptome data used in this study.

This study investigated the in vitro adhesion and biofilm formation capacity of Candida parapsilosis and Kodamaea ohmeri isolates from onychomycoses of HIV/AIDS patients and also established the antifungal sensitivity profiles of these isolates. Onychomycosis in HIV/AIDS patients showed a high prevalence of emerging yeasts, among which C. parapsilosis complex species and K. ohmeri were the most frequent. Three C. parapsilosis sensu stricto isolates and two C. orthopsilosis isolates were resistant to amphotericin B and 83% were resistant to terbinafine. All isolates adhered to stainless steel and siliconized latex surfaces, and carbohydrates intensified adhesion of all isolates. Isolates adhered to keratinous nail and 50% formed biofilms with strong intensity. In multispecies or polymicrobial biofilms, C. albicans and Staphylococcus aureus regulated the formation of biofilms, decreasing the number of non-albicans species and of Candida spp. cells, respectively. Nonetheless, the presence of C. parapsilosis sensu stricto or K. ohmeri in mixed biofilms increased the number of viable C. albicans cells. The isolation of emerging yeast species from onychomycosis with high adhesion capacity to medical devices, with many being good biofilm producers, is a result that should be considered relevant in clinical practice. In addition, half of the isolates resistant to at least one of the tested antifungals or were susceptible in a dose-dependent manner, which corroborates the infectious capacity and viability of these isolates.

The supplementary material and research data for the article - The effects of novel heme oxygenase inhibitors on the growth of Pseudomonas aeruginosa 1. General Synthetic Procedures 2. The determination of DMSO toxicity on P.aeruginosa growth 3. Determination of the MIC50 values of the compounds on PAO1

Fig S1. Cloning and Expression of recombinant proteins Rv2627 and Rv2628 a) Positive plasmid PCR of gene Rv2627 (L1) with product size 1242bp and gene Rv2628 (L2) with product size 363bp and M- Marker. b) SDS PAGE of purified Rv2627 and Rv2628 protein in native condition. M-Marker and L1- Rv2627 and L2- Rv2628 purified protein. The molecular weight of Rv2627 was 46.5 kDa and Rv2628 was 13.1 kDa. c) Western blot of purified Rv2627 and Rv2628. M-Marker and L1- Rv2627 and L2- Rv2628 purified protein. Fig S2. Gating strategies used for acquiring and analysis of CD3+ cells using labeled un-stimulated and stimulated cells and their FMO controls a) CD4+ IFN-γ+, b) CD4+CD45RA+, c) CD4+CD45RO+, d) CD8+IFN-γ+, e) CD8+CD45RA+, f) CD8+ CD45RO+ and g) Treg (CD4+CD25+FoxP3+). Fig S3. Basal percentage of different T cell markers in the three sample population of Normal (n=20), Contacts (n=20) and PTB patients (n=20). The normal individuals had highest percentage of CD4+CD45RA+, CD4+CD45RO+, CD8+CD45RA+, CD8+CD45RO+ cells as compared to that of contacts and patients whereas the PTB patients had highest percentage of Treg cells. Statistical differences between samples were calculated by the Mann-Whitney U test for unpaired samples. Values were shown as mean ± SD, *P < 0.05, **P < 0.01, ***P < 0.001 and ****P < 0.0001. Fig S4. Basal Cytokine production profile in PBMC of three sample population of Normal (n=15), Contacts (n=15) and PTB patients (n=15). The contacts had higher basal level of cytokines like IFN-γ and IL-2 whereas PTB patients had higher basal levels of cytokines like IL-4 and TGF-. Statistical differences between samples were calculated by the Mann-Whitney U test for unpaired samples. Values were shown as mean ± SD, *P < 0.05, **P < 0.01 and ***P < 0.001. Fig S5. Rv2627 and Rv2628 skew the immune response towards Th1 in both contacts and patients by increasing the IFN-γ production In contacts, IFN-γ/IL-4 ratio increased significantly only for Rv2628, whereas in PTB samples the ratio increased for both the proteins. Statistical differences between unstimulated and stimulated samples were calculated by the Wilcoxon rank sum test for paired samples. Values were shown as mean ± SD, *P < 0.05.

Cell viability assay to check the death rate of hepatocytes.

The assay that were performed in the MS.

These articles are related to the submitted article.

The GapC protein of Staphylococcus aureus is a surface protein that is highly conserved among Staphylococcus strains, and it can induce protective humoral immune responses. However, B-cell epitopes in S. aureus GapC have not been reported. In this study, we generated a monoclonal antibody (mAb2A9) targeting S. aureus GapC. Through a passive immunity test, mAb2A9 was shown to partially protect mice against S. aureus infection. We screened the motif 236PVATGSLTE243 that is recognized by mAb2A9 using a phage-display system. The motif sequence exactly matched amino acids 236–243 of the S. aureus GapC protein. Then, we identified the key amino acids in the motif using site-directed mutagenesis. Site-directed mutagenesis revealed that residues P236, G240, L242, and T243 formed the core of the 236PVATGSLT243 motif. In addition, this epitope was proven to be located on the surface of S. aureus, and it induced a protective humoral immune response against S. aureus infection in immunized mice. Overall, our results characterized a conserved B-cell epitope, which will be an attractive target for designing effective epitope-based vaccines against S. aureus infection.