Journal: mBio

Article Title: Phylogenetic analysis of pathogenic algae reveals lineage-dependent patterns of phagocytosis

doi: 10.1128/mbio.00498-25

Figure Lengend Snippet: J774A.1 cells, but not HMDMs, phagocytose P. miyajii most efficiently compared with other pathogenic, human-associated species. J774A.1 cells and HMDMs were cultured as described in Materials and Methods, seeded in 24-well plates at 1 × 10 5 /well or 5 × 10 5 /well, respectively, and incubated at 37°C and 5% CO 2 . The next day, cells were challenged with P. bovis (HP3, HP40, or HP41), P. ciferii (HP2), P. blaschkeae (HP4), P. cutis (HP28), P. miyajii (HP27), or P. wickerhamii (HP50 or HP52) at an MOI = 3 (i.e., 3 algal cells:1 macrophage) and imaged every minute for 1 hour on either a Nikon Eclipse Ti microscope or a Zeiss Axio Observer microscope using a 20× objective lens. Data are expressed as the mean ± SEM of at least two independent, biological experiments, where n = at least 50 macrophages analyzed per frame of view per technical replicate per biological replicate for each strain and cell type. Videos were exported to and analyzed in Fiji for phagocytic index, defined as the percentage of macrophages that take up at least one algal cell by the end of the imaging period. ( A ) Phagocytic index for cattle-associated species exposed to J774A.1 cells. ( B ) Phagocytic index for human-associated species exposed to HMDMs. Statistical significance was assessed by an unpaired t -test, where P < 0.05 was considered significant. ns = not significant, ** P < 0.01.

Article Snippet: To determine whether the sub-lineages identified might have different interactions with mammalian immunity, algal cells were used to infect J774A.1 murine macrophages (the European Collection of Authenticated Cell Cultures) or human monocyte-derived macrophages (HMDMs).

Techniques: Cell Culture, Incubation, Microscopy, Imaging

Journal: mBio

Article Title: Phylogenetic analysis of pathogenic algae reveals lineage-dependent patterns of phagocytosis

doi: 10.1128/mbio.00498-25

Figure Lengend Snippet: Pathogenic and non-pathogenic cattle-associated species and strains of Prototheca are phagocytosed most efficiently by both J774A.1 cells and HMDMs. J774A.1 cells and HMDMs were cultured as described in Materials and Methods, seeded in 24-well plates at 1 × 10 5 /well or 5 × 10 5 /well, respectively, and incubated at 37°C and 5% CO 2 . The next day, cells were challenged with P. bovis (HP3, HP40, or HP41), P. ciferii (HP2), P. blaschkeae (HP4), P. cookei (HP32), P. cutis (HP28), P. miyajii (HP27), P. wickerhamii (HP50 or HP52), P. paracutis (HP31), P. tumulicola (HP29), P. moriformis (HP51), P. xanthoriae (HP53 or HP54), A. protothecoides (HA1 or HA2), A. symbiontica (HA5), or one of two unidentified Auxenochlorella spp. (HA6 or HA7) at an MOI = 3 (i.e., 3 algal cells:1 macrophage) or MOI = 1 for some strains, and imaged every minute for one hour on either a Nikon Eclipse Ti microscope or a Zeiss Axio Observer microscope using a 20× objective lens. Data are expressed as the mean ± SEM of at least two independent, biological experiments, where n = at least 50 macrophages analyzed per frame of view per technical replicate per biological replicate for each strain and cell type. Videos were exported to and analyzed in Fiji for phagocytic index ( A and B ), defined as the percentage of macrophages that take up at least one algal cell by the end of the imaging period, and mean uptake ( C and D ), defined as the mean number of algal cells taken up by a single macrophage by the end of the imaging period. Statistical significance was assessed by one-way ANOVA followed by Tukey’s multiple comparisons test, where P < 0.05 was considered significant. Group 1: cattle-associated species. Group 2: human-associated species. Group 3: true environmental species. Group 4: P. xanthoriae and Auxenochlorella spp.

Article Snippet: To determine whether the sub-lineages identified might have different interactions with mammalian immunity, algal cells were used to infect J774A.1 murine macrophages (the European Collection of Authenticated Cell Cultures) or human monocyte-derived macrophages (HMDMs).

Techniques: Cell Culture, Incubation, Microscopy, Imaging

Journal: mBio

Article Title: Phylogenetic analysis of pathogenic algae reveals lineage-dependent patterns of phagocytosis

doi: 10.1128/mbio.00498-25

Figure Lengend Snippet: Macrophage migratory dynamics vary to different extents across sub-lineages within the AHP lineage. J774A.1 cells and HMDMs were cultured as described in Materials and Methods, seeded in 24-well plates at 1 × 10 5 /well or 5 × 10 5 /well, respectively, and incubated at 37°C and 5% CO 2 . The next day, cells were challenged with P. bovis (HP3, HP40, or HP41), P. ciferii (HP2), P. blaschkeae (HP4), or P. cookei (HP32) at an MOI = 3 (i.e., 3 algal cells:1 macrophage) and imaged every minute for 1 hour on either a Nikon Eclipse Ti microscope or a Zeiss Axio Observer microscope using a 20× objective lens. Data are expressed as the mean ± SEM of at least three independent, biological experiments, where n = at least 50 macrophages analyzed per frame of view per technical replicate per biological replicate for each strain and cell type. Videos were exported to and analyzed in Fiji for migratory behavior parameters. ( A and E ) Data show mean displacement (μm). ( B and F ) Data show mean speed (μm/s). ( C and G ) Data show linearity of forward progression (fraction of 1). ( D and H ) Data show mean directional change rate (rad/sec). Statistical significance was assessed by one-way ANOVA followed by Tukey’s multiple comparisons test, where P < 0.05 was considered significant. Group 1: cattle-associated species. Group 2: human-associated species. Group 3: true environmental species. Group 4: P. xanthoriae and Auxenochlorella spp.

Article Snippet: To determine whether the sub-lineages identified might have different interactions with mammalian immunity, algal cells were used to infect J774A.1 murine macrophages (the European Collection of Authenticated Cell Cultures) or human monocyte-derived macrophages (HMDMs).

Techniques: Cell Culture, Incubation, Microscopy

Journal: Metabolism Open

Article Title: A systematic review on the effects of Echinacea supplementation on cytokine levels: Is there a role in COVID-19?

doi: 10.1016/j.metop.2021.100115

Figure Lengend Snippet: Characteristics of the in vitro and ex vivo studies included.

Article Snippet: Matthias, 2007 [ ] , MediHerb Research Laboratories, Queensland, Australia , Mouse macrophage cell line , LPS (0.1 μg/mL) or PMA (2 nM) , Alkylamide 1. (2E)-N-isobutylundeca-2-ene-8,10-diynamide; Alkylamide 2. (2E,4E,8Z,10Z)-N-isobutyldodeca-2,4,8,10-tetraenamide.; An ethanolic extract ( Echinacea Premium Liquid; EPL) of E. purpurea (300 mg/mL), E. angustifolia (200 mg/mL) roots and EPL alkylamide fraction (EPL AA) was separated from caffeic acid fraction and cichoric acid , Alkylamides concentration 0.2 ng/mL; cichoric acid concentration 0.8 ng/mL DURATION: 4 and 20 h , Unstimulated cells , Decreased production: TNF-α , 3.

Techniques: In Vitro, Ex Vivo, Infection, Control, Negative Control, Positive Control, Expressing, Solvent, Isolation, Concentration Assay, High Performance Liquid Chromatography, Gene Expression, Size-exclusion Chromatography, Synthesized, Extraction, Saline, Virus, Mouse Assay, Clinical Proteomics, Cell Culture, Derivative Assay, Purification, Chromatography, Colony-forming Unit Assay, Cytokine Assay, Irradiation