Journal: Vaccines

Article Title: Systemic and Lower Respiratory Tract Immunity to SARS-CoV-2 Omicron and Variants in Pediatric Severe COVID-19 and Mis-C

doi: 10.3390/vaccines10020270

Figure Lengend Snippet: Neutralizing antibody responses in plasma and endotracheal aspirates of pediatric COVID-19 and MIS-C against SARS-CoV-2 WA1/2020 strain and variant strains. SARS-CoV-2 neutralizing antibody titers of plasma ( A ) and endotracheal aspirates ( B ) from 5 COVID-19 and 5 MIS-C pediatric patients against SARS-CoV-2 WA1/2020 strain, VOIs; Epsilon, Iota, Kappa, and VOCs; Alpha, Beta, Gamma, Delta and Omicron variant by pseudovirion neutralization assay (PsVNA). The assay of each sample was performed in duplicate to determine the 50% neutralization titer (PsVNA50). Each data point represents an individual sample (circles) and indicates the 50% neutralization titer obtained with each sample against the indicated pseudovirus. Individual COVID-19 plasma samples are shown as circles, while MIS-C plasma samples are shown as triangles in panel A. In panel B, the endotracheal aspirates samples from pediatric COVID-19 are shown as squares, while MIS-C are shown as inverted triangles. The heights of the bars and the numbers over the bars indicate the geometric mean titers, and the whiskers indicate 95% confidence intervals. The horizontal dashed line indicates the limit of detection for the neutralization assay (PsVNA50 of 20). The fold-change indicates the average decrease in neutralization titer of the indicated variants as compared with that of the SARS-CoV-2 WA1/2020 virus. The raw data and information regarding the samples for each pediatric participant (age and 50% neutralization titers against various SARS-CoV-2 strains) are summarized in . Differences in neutralization titers between SARS-CoV-2 strains were analyzed by lme4 and emmeans packages in R using Tukey’s pairwise multiple comparison test and the p -values are shown.

Article Snippet: For comparisons between the SARS-CoV-2 strains (factor variable), pairwise comparisons were extracted using ‘emmeans’ and Tukey-adjusted p values were used for denoting significance to reduce Type 1 error due to multiple testing in R (RStudio version 1.1.463).

Techniques: Clinical Proteomics, Variant Assay, Neutralization, Virus, Comparison

Journal: PLOS Pathogens

Article Title: Aspergillus fumigatus transcription factor ZfpA regulates hyphal development and alters susceptibility to antifungals and neutrophil killing during infection

doi: 10.1371/journal.ppat.1011152

Figure Lengend Snippet: 2-day post fertilization wild-type larvae with fluorescent macrophages (GFP) and neutrophils (BFP) were infected with RFP-expressing WT CEA10, Δ zfpA , or OE:: zfpA strains. Larvae were imaged with confocal microscopy at 24, 48, 72, and 96 hours post infection (hpi). (A) Representative images of fungal growth and immune cell recruitment in the same larva at 24–96 hpi. Images represent maximum intensity projections of z-stacks. Scale bar = 50 μm. (B) Mean percentage of larvae with germinated spores at 24–96 hpi. Dots and error bars represent mean+s.d. (C) 2D fungal (RFP) area at 24–96 hpi. (D) 2D neutrophil (BFP) area 24–96 hpi. (E) 2D macrophage (GFP) area at 24–96 hpi. Bars in (C-E) represent lsmeans+s.e.m. Results represent data pooled from 4 independent experiments. n = 45–48 larvae per condition. p values calculated by ANOVA with Tukey’s multiple comparisons. * p <0.05, ** p <0.01.

Article Snippet: 2D area of fungal growth, neutrophils, and macrophages represent least-squared adjusted means±standard error of the mean (LSmeans±s.e.m.) and were compared using ANOVA with Tukey’s multiple comparisons (RStudio).

Techniques: Infection, Expressing, Confocal Microscopy

Journal: PLOS Pathogens

Article Title: Aspergillus fumigatus transcription factor ZfpA regulates hyphal development and alters susceptibility to antifungals and neutrophil killing during infection

doi: 10.1371/journal.ppat.1011152

Figure Lengend Snippet: (A) Susceptibility of WT CEA10, Δ zfpA , and OE:: zfpA to 0.1 and 0.25 μg/ml voriconazole (VOR). 10 4 spores were point-inoculated on solid GMM with voriconazole or DMSO. Bars represent mean±s.d. of relative colony diameter of 4 plates per condition. p values calculated by ANOVA with Tukey’s multiple comparisons. *** p <0.001, **** p <0.0001. (B) Survival analysis of infected Rac2D57N larvae (neutrophil-deficient) bathed in 0.1 μg/ml voriconazole or 0.001% DMSO. Average spore dose injected: WT CEA10 = 39, Δ zfpA = 35, OE:: zfpA = 37. Results represent pooled data from 3 independent experiments. n = 44–66 larvae per condition. p values and hazard ratios calculated by Cox proportional hazard regression analysis.

Article Snippet: 2D area of fungal growth, neutrophils, and macrophages represent least-squared adjusted means±standard error of the mean (LSmeans±s.e.m.) and were compared using ANOVA with Tukey’s multiple comparisons (RStudio).

Techniques: Infection, Injection

Journal: PLOS Pathogens

Article Title: Aspergillus fumigatus transcription factor ZfpA regulates hyphal development and alters susceptibility to antifungals and neutrophil killing during infection

doi: 10.1371/journal.ppat.1011152

Figure Lengend Snippet: (A) Susceptibility of WT CEA10, Δ zfpA , and OE:: zfpA to 0.25, 0.5, 1, and 8 μg/ml caspofungin (CSP). 10 4 spores were point-inoculated on solid GMM with caspofungin or DMSO. Bars represent mean±s.d. of colony diameter at 4 days post inoculation of 4 plates per condition. (B) Susceptibility of WT CEA10, Δ zfpA , and OE:: zfpA to 0.25, 0.5, 1, and 8 μg/ml micafungin (MCF). 10 4 spores were point-inoculated on solid GMM with micafungin or DMSO. Bars represent mean±s.d. of colony diameter at 4 days post inoculation of 4 plates per condition. p values calculated by ANOVA with Tukey’s multiple comparisons. (C) Images represent calcofluor white (CFW) staining of WT CEA10, Δ zfpA , and OE:: zfpA following overnight exposure to 1 μg/mL caspofungin (CSP) or DMSO. CFW staining is represented by cyan and cytoplasmic RFP signal is shown in magenta. Scale bar = 50 μm. (D) Mean gray value of CFW signal following DMSO or caspofungin treatment. Bars represent mean±s.e.m of 3 independent experiments. n = 24–30 hyphae per condition. (E) Experimental setup for chitin stimulation with CaCl 2 /CFW. Spores were incubated for 8 h at 37°C or until germination in liquid GMM or liquid GMM supplemented with 0.2 M CaCl 2 and 100 μg/mL CFW. After germination, media was replaced for GMM + 1 μg/mL caspofungin or DMSO and hyphae were incubated for an additional 12 h before detecting PrestoBlue viability reagent signal in a plate reader. (F) Bars represent mean±s.d. of relative fungal viability following caspofungin exposure. Relative viability was calculated by normalizing the mean signal of caspofungin-treated wells to the mean signal of DMSO-treated wells. All experiments included 5 wells/condition. Data are pooled from 3 independent experiments. p values calculated by ANOVA with Sidak’s multiple comparisons. * p <0.05, ** p <0.01, *** p <0.001, **** p <0.0001.

Article Snippet: 2D area of fungal growth, neutrophils, and macrophages represent least-squared adjusted means±standard error of the mean (LSmeans±s.e.m.) and were compared using ANOVA with Tukey’s multiple comparisons (RStudio).

Techniques: Staining, Incubation