Journal: Toxins

Article Title: A Functional ClpXP Protease is Required for Induction of the Accessory Toxin Genes, tst, sed, and sec

doi: 10.3390/toxins12090553

Figure Lengend Snippet: SED and TSST-1 protein levels are reduced in supernatants derived from cells lacking ClpXP activity. ( A ) Growth curves of the Sa564 wild-type and mutant strains in TSB at 37 °C. (•) SA564 wild-type, (□) SA564Δ clpX , and ( ▲ ) SA564Δ clpP . Average values and standard deviations of three independent biological replicates ( n = 3) are shown. ( B ) Normalized SED production of the Sa564 wild-type and mutant strains. Dark-gray filled bars—SA564 wild-type; white bars—SA564Δ clpX, and light-gray filled bars—SA564Δ clpP . Average values and standard deviations of three independent biological replicates ( n = 3) are shown. ( C ) Normalized TSST production of the Sa564 wild-type and mutant strains. Dark-gray filled bars—SA564 wild-type, white bars—SA564Δ clpX , and light-gray filled bars—SA564Δ clpP . Average values and standard deviations of two independent biological replicates ( n = 2) are shown.

Article Snippet: In this study, affinity-purified antibodies (IgG), sheep anti SED, or TSST coating IgG, and biotinylated sheep anti SED or TSST detection IgG (Toxin Technology Inc., Sarasota, FL, USA) were used.

Techniques: Derivative Assay, Activity Assay, Mutagenesis

Journal: Toxins

Article Title: A Functional ClpXP Protease is Required for Induction of the Accessory Toxin Genes, tst, sed, and sec

doi: 10.3390/toxins12090553

Figure Lengend Snippet: Diminished levels of the Rot repressor in cells with decreased transcription of the sed, sec , and tst genes. The cellular levels of the Rot transcriptional regulator were determined in the samples of SA564 wild-type, SA564∆ clpP , and SA564∆ clpX cells derived from the same cultures, as used for the RNA extraction (see the Northern blotting analysis presented in ). Accordingly, proteins were extracted from cells in late-exponential (OD 600 = 1.5), early post-exponential growth phase (OD 600 = 3.0), or late post-exponential growth phase (OD 600 = 6.0), and the extracted proteins were separated by SDS–PAGE, blotted onto a PVDF membrane, and probed with anti-Rot antibody. The protein ladder is shown to the left. The upper band resulting from non-specific binding of the Rot antibody to an unknown cellular protein visualizes equal loading.

Article Snippet: In this study, affinity-purified antibodies (IgG), sheep anti SED, or TSST coating IgG, and biotinylated sheep anti SED or TSST detection IgG (Toxin Technology Inc., Sarasota, FL, USA) were used.

Techniques: Derivative Assay, RNA Extraction, Northern Blot, SDS Page, Membrane, Binding Assay

Journal: Scientific Reports

Article Title: Antibody and Memory B Cell Responses in Hepatitis E Recovered Individuals, 1–30 Years Post Hepatitis E Virus Infection

doi: 10.1038/s41598-019-40603-9

Figure Lengend Snippet: Clinical characteristics of study groups. Data are shown as median (range); NA: Not applicable.

Article Snippet: On day 3, the plates were washed with PBS containing 0.5% (v/v) Tween-20 and blocked with RPMI-1640 + 10%FBS at 37 °C for 3–4 h. PBMCs were harvested and 1 × 10 5 unstimulated/stimulated cells per well were added in triplicates to rORF2p-coated wells, while 1 × 10 5 stimulated cells per well were added to anti-IgG-coated wells, along with anti-HEV IgG-biotinylated detection antibody (1:100; Mabtech, Sweden) and incubated at 37 °C.

Techniques:

Journal: Scientific Reports

Article Title: Antibody and Memory B Cell Responses in Hepatitis E Recovered Individuals, 1–30 Years Post Hepatitis E Virus Infection

doi: 10.1038/s41598-019-40603-9

Figure Lengend Snippet: Anti-HEV IgG antibody positivity. Plasma of all study participants was used for detection of anti-HEV IgG antibodies. In positive samples, IgG avidity indices were determined. The dots represent individual values and bars represent median values. ( a ) Anti-HEV IgG positivity in acute hepatitis E patients (n = 31) and hepatitis E recovered individuals (n = 35). Dashed line shows the cut off value for the assay ( b ) Anti-HEV IgG antibodies in all hepatitis E recovered individuals 1–30 years post infection ( c ) IgG avidity indices of all acute hepatitis E patients and hepatitis E recovered individuals. An index <40% was considered as low, >60% as high and 40–60% as equivocal and are represented by the dotted and dashed lines.

Article Snippet: On day 3, the plates were washed with PBS containing 0.5% (v/v) Tween-20 and blocked with RPMI-1640 + 10%FBS at 37 °C for 3–4 h. PBMCs were harvested and 1 × 10 5 unstimulated/stimulated cells per well were added in triplicates to rORF2p-coated wells, while 1 × 10 5 stimulated cells per well were added to anti-IgG-coated wells, along with anti-HEV IgG-biotinylated detection antibody (1:100; Mabtech, Sweden) and incubated at 37 °C.

Techniques: Clinical Proteomics, Infection