Journal: Animals : an Open Access Journal from MDPI

Article Title: Procyanidin-B1-Enriched Cyperus esculentus Extract Regulates Anti-Inflammatory Pathways in Chicken Macrophages Cells Through Transcription Factor STAT2 and HIF1A

doi: 10.3390/ani15233469

Figure Lengend Snippet: FLAVER analysis results of the transcriptome data from chicken layers treated with procyanidin B1. The dots represent the Log10(FDR) values from the FLAVER analysis results, with green indicating a positive correlation between transcription factor binding sites and target gene expression, and blue indicating a negative correlation. Results of the significant transcription factors are shown in plots ( A – C ) arranged in alphabetical order.

Article Snippet: Procyanidin B1 standard ( B21616 , CAS No. 20315-25-7, ≥95%) was obtained from Shanghai Yuanye Bio-Technology Co., Ltd. (Shanghai, China).

Techniques: Binding Assay, Targeted Gene Expression

Journal: iScience

Article Title: Red blood cell signaling is functionally conserved in Plasmodium invasion

doi: 10.1016/j.isci.2024.111052

Figure Lengend Snippet: Properties of signaling inhibitors used in the merozoite invasion inhibition assays and RBC cAMP-Ca 2+ signaling assays

Article Snippet: , NF449 , – , Santa Cruz Biotechnology , Gαs , Guo et al. .

Techniques: Inhibition, Membrane

Journal: iScience

Article Title: Red blood cell signaling is functionally conserved in Plasmodium invasion

doi: 10.1016/j.isci.2024.111052

Figure Lengend Snippet: RH5-induced cAMP increase promotes RBC Ca 2+ influx Recombinant rRH5 was purified and used for all the assays. (A) Fluorescence images of RBC Ca 2+ . Fresh RBCs labeled with Fluo-4a.m. were incubated in the absence (i, RBC only) and presence of rRH5 (ii, rRH5) and presence of BIM46187 (iii, BIM46187+rRH5), 2′,5′-ddA (iv, 2′,5′-ddA + rRH5), PKA (14–22) (v, PKA (14–22)+rRH5) or verapamil (vi, Verapamil+rRH5) prior to rRH5 binding for 10 min. Fluorescence (left) and bright field (right) images were taken under 20× objective lens. Scale bar = 50 μm. (B) RBC Ca 2+ measurement by using fluorescence plate reader. The Ca 2+ signal intensity of the samples obtained from (A) were also measured. Dynamics of RBC cytosolic Ca 2+ level was measured over 600 s ΣΔF(t) which reflects the cumulative change in RBC cytosolic Ca 2+ was plotted as a bar chart. Experimental data presented as mean ± s.e.m, n = 3. (C) cAMP inhibition immunoassay of resealed RBCs (rRBCs). To avoid the off-target effect on the permeable inhibitors mentioned above, membrane-impermeable inhibitors ( Table 1 ) were loaded into rRBCs. The dynamics of rRBC cAMP level was measured upon rRH5 binding for 2 min in the absence (rRBC) and presence of impermeable inhibitors listed in Table 1 . The cAMP reading in HA-loaded rRBCs was used as an internal positive control (HA). ∗∗∗ p = 0.01, ### p = 0.01 and $$$ p = 0.02 indicate the significant difference on fold change in rRBC cAMP level upon rHR5 binding in the presence of GPA, NF449 or 2′- d -3′-AMP compared to that in the HA-loaded rRBC, respectively (color in green). Whereas no significant inhibition effect was seen in Rp-8-OH-cAMPS, PKA (6–22) or N-methyl Vera with p > 0.05 including p = 0.17, p = 0.06 and p = 0.20 (color in red). Experimental data presented as mean ± s.e.m, n = 3. (D) rRBC Ca 2+ measurement by using fluorescence plate reader. Fluo-4a.m. labeled rRBCs were incubated with rRH5 protein in the absence (rRH5) and presence of impermeable inhibitors GPA, NF449, 2′- d -3′-AMP, Rp-8-OH-cAMPS, PKA (6–22) or N-methyl Vera, respectively. HA-loaded rRBC (HA) was used as an internal positive control. Dynamics of RBC cytosolic Ca 2+ level was measured over 600 s ΣΔF(t) which reflects the cumulative change in RBC cytosolic Ca 2+ was plotted as a bar chart. Experimental data presented as mean ± s.e.m, n = 3. Statistical comparison was done using one-way ANOVA.

Article Snippet: , NF449 , – , Santa Cruz Biotechnology , Gαs , Guo et al. .

Techniques: Recombinant, Purification, Fluorescence, Labeling, Incubation, Binding Assay, Inhibition, Membrane, Positive Control, Comparison

Journal: iScience

Article Title: Red blood cell signaling is functionally conserved in Plasmodium invasion

doi: 10.1016/j.isci.2024.111052

Figure Lengend Snippet: Live video microscopy of P. falciparum and P. knowlesi merozoite invasion (A) Representative snapshots were taken from time-lapse live video microscopy of P. falciparum 3D7 merozoite invasion in the Fluo-4a.m. labeled human resealed RBCs in the absence (rRBC) (i) and presence of HA peptide (HA) (ii). Merozoite attaches to the RBC after the schizont rupture and reorients followed by Ca 2+ influx that spreads throughout the entire resealed RBC. The Ca 2+ influx causes a massive deformation, and the signal starts to decrease when echinocytosis happens ( and ). Representative video snapshots were also taken for both 3D7 invasion in the Fluo-4a.m. labeled rRBCs in the presence of (iii) GPA, (iv) NF449, (v) 2′- d -3′-AMP, (vi) Rp-8-OH-cAMPS, (vii) PKA (6–22) and (viii) N-methyl Vera ( , , , , , ). (B) Snapshots of P. knowlesi A1-H.1 merozoite invasion in the Fluo-4a.m. labeled human resealed RBCs in the presence of HA peptide (i), GPA (ii) and N-methyl Vera (iii). Merozoite attaches to the rRBC loaded with HA peptide (i) after the schizont rupture and reorients followed by Ca 2+ influx that spreads throughout the entire resealed RBC. The Ca 2+ influx causes a massive deformation, and the signal starts to decrease when echinocytosis happens . In snapshots of (A-iii-viii) and (B-ii - iii), the merozoite released from a mature schizont attaches and apically reorients, but it is unable to penetrate the RBC with no Ca 2+ influx ( and ). Dynamics of rRBC Ca 2+ signals was measured. Ca 2+ fluorescence intensity was indicated next to each of the snapshot. Relative time was shown in seconds. Scale bar = 5 μm. (C) rRBC Ca 2+ measurement by using fluorescence plate reader during P. falciparum and P. knowlesi invasion. Fluo-4a.m. labeled rRBCs were incubated with 3D7 ( P.f ) and A1-H.1 ( P.k ) merozoites in the absence (rRBC) and presence of inhibitors GPA, NF449, 2′- d -3′-AMP, Rp-8-OH-cAMPS, PKA (6–22) or N-methyl Vera, respectively. HA-loaded resealed RBC (HA) was used as an internal positive control. Dynamics of RBC cytosolic Ca 2+ level was measured over 600 s ΣΔF(t) which reflects the cumulative change in RBC cytosolic Ca 2+ was plotted as a bar chart. Experimental data presented as mean ± s.e.m, n = 3. (D) cAMP and Ca 2+ signaling inhibitors impede junction formation. 3D7 schizonts were incubated with rRBC in the absence and presence of inhibitors ( Table 1 ) under Cyto D treatment. HA-loaded rRBCs were used as an internal control. Junction-arrested merozoites were counted microscopically. % junction-arrested merozoites of tested samples from 3 biological replicates were plotted. All inhibitors effectively blocked junction formation. Experimental data presented as mean ± s.e.m, n = 3.

Article Snippet: , NF449 , – , Santa Cruz Biotechnology , Gαs , Guo et al. .

Techniques: Microscopy, Labeling, Fluorescence, Incubation, Positive Control, Control

Journal: iScience

Article Title: Red blood cell signaling is functionally conserved in Plasmodium invasion

doi: 10.1016/j.isci.2024.111052

Figure Lengend Snippet: Synergistic effect of cAMP-Ca 2+ signaling inhibitors block junction formation during merozoite invasion (A) Inhibition heat plots of 3D7 merozoite invasion into rRBCs. Resealed RBCs were pre-loaded with dual combinations of Ca 2+ channel blocker (N-methyl Vera) and other cAMP signaling inhibitors including GPA (i, red), NF449 (ii, yellow), 2′- d -3′-AMP (iii, blue), Rp-8-OH-cAMPS (iv, green), and PKA (6–22) (v, purple). All inhibitors were loaded in rRBCs at about 4-fold increasing concentrations including N-methyl Vera shown in each table vertically, while the other inhibitors shown horizontally in color matrices. Colors in the dose-response matrices from the darkest to white indicate different levels of invasion inhibitions from 0% to 100% obtained from Figure S3 . (B) Isobologram plot of five different checkerboard invasion inhibition data from Figure S4 . The range of Fractional Inhibitory Concentration (FIC) values surrounding the black dotted line with arrow in black from 0 to 0.5 represents a synergistic isobole. Data points below this line indicate potent synergism, while the data points above this line indicate partial synergism. Inset: zoomed plot at 0.15 of both x and y axis for better visualisation.

Article Snippet: , NF449 , – , Santa Cruz Biotechnology , Gαs , Guo et al. .

Techniques: Blocking Assay, Inhibition, Concentration Assay

Journal: iScience

Article Title: Red blood cell signaling is functionally conserved in Plasmodium invasion

doi: 10.1016/j.isci.2024.111052

Figure Lengend Snippet:

Article Snippet: , NF449 , – , Santa Cruz Biotechnology , Gαs , Guo et al. .

Techniques: Recombinant, Giemsa Stain, Saline, Lysis, Membrane, Fluorescence, Blocking Assay, Purification, Protein Purification, cAMP Assay, Competitive ELISA, Plasmid Preparation, Software, Imaging

Journal: Sensors (Basel, Switzerland)

Article Title: Raman-Activated, Interactive Sorting of Isotope-Labeled Bacteria

doi: 10.3390/s24144503

Figure Lengend Snippet: ( A ) Raman mean spectra of single bacteria ( E. coli and M. lylae ) without isotope labeling (control, blue) and with labeling with D (red) or 13 C (green). ( B ) LD plot for the discrimination of different labeled bacteria with the subsequent loading vectors ( C ).

Article Snippet: For this study, the bacterial strains Escherichia coli ( E. coli ) DSM 498 and Micrococcus lylae ( M. lylae ) DSM 20,318 purchased from the German Collection of Microorganisms and Cell Cultures (DSMZ, Braunschweig, Germany) were used.

Techniques: Bacteria, Quantitative Proteomics, Control, Labeling

Journal: Sensors (Basel, Switzerland)

Article Title: Raman-Activated, Interactive Sorting of Isotope-Labeled Bacteria

doi: 10.3390/s24144503

Figure Lengend Snippet: Number of bacterial cells exhibiting the isotope signal after washing.

Article Snippet: For this study, the bacterial strains Escherichia coli ( E. coli ) DSM 498 and Micrococcus lylae ( M. lylae ) DSM 20,318 purchased from the German Collection of Microorganisms and Cell Cultures (DSMZ, Braunschweig, Germany) were used.

Techniques: