Journal: Plants

Article Title: Phenolics Profiling by HPLC-DAD-ESI/MS n of the Scientific Unknown Polygonum hydropiperoides Michx. and Its Antioxidant and Anti-Methicillin-Resistant Staphylococcus aureus Activities

doi: 10.3390/plants12081606

Figure Lengend Snippet: Minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) values obtained for the hexane (HE-Ph), ethyl acetate (EAE-Ph), and ethanolic (EE-Ph) extracts from the aerial parts of Polygonum hydropiperoides , and ampicillin (AMP) and chloramphenicol (CHL) against the bacterial strains tested.

Article Snippet: Nine American Type Culture Collection (ATCC ® ) bacterial strains of methicillin-sensitive Staphylococcus aureus subsp. aureus Rosenbach (ATCC 6538, ATCC 25923 and ATCC 29213) (MSSA), Escherichia coli (Migula) Castellani and Chalmers (ATCC 10536 and ATCC 25922), Salmonella enterica subsp. enterica ( ex Kauffmann and Edwards) Le Minor and Popoff serovar Choleraesuis (ATCC 10708), Salmonella enterica subsp. enterica ( ex Kauffmann and Edwards) Le Minor and Popoff serovar Typhimurium (ATCC 13331), and Pseudomonas aeruginosa (Schroeter) Migula (ATCC 9027 and ATCC 27853), described as requested by ATCC ® , were assayed.

Techniques: Concentration Assay

Journal: STAR Protocols

Article Title: Click-chemistry-based protocol to quantitatively assess fatty acid uptake by Mycobacterium tuberculosis in axenic culture and inside mouse macrophages

doi: 10.1016/j.xpro.2023.102062

Figure Lengend Snippet:

Article Snippet: Mus musculus : L929 cell line, Passage 5–15 , ATCC , Cat#CCL-1.

Techniques: Recombinant, Staining, Electron Microscopy, Modification, Software, Imaging, Cell Culture, Flow Cytometry, Microscopy

Journal: eLife

Article Title: Maternal obesity blunts antimicrobial responses in fetal monocytes

doi: 10.7554/eLife.81320

Figure Lengend Snippet: ( A ) Experimental design for in vitro bacterial and viral stimulation. Purified monocytes were cultured in the presence/absence of E. coli or RSV for 16 hr. Cell pellets were used for bulk RNA-Seq analyses and supernatants were used for Luminex analyses of secreted cytokines and chemokines. ( B ) Bubble plot of key secreted factors significantly different following E. coli infection. The size of the bubble represents the quantity of the secreted analyte (log-transformed) whereas color represents statistical significance relative to no stimulation controls. Statistically significant analytes between lean and obese groups are highlighted with # - p<0.05. ( C ) Venn diagram (right) and corresponding functional enrichment (left) of genes upregulated with E. coli infection in lean and obese groups (Green denotes common DEG, blue DEG unique to lean group, and yellow DEG unique to the obese group) using metascape. The length of the bar indicates the number of genes in each gene ontology (GO) term. Color represents the statistical significance of each GO term. ( D ) Heatmap comparing fold changes of the genes upregulated in both groups (77 genes) that mapped to GO terms ‘myeloid cell differentiation’, ‘inflammatory response to antigenic stimulus’, ‘leukocyte apoptotic process’, ‘regulation of leukocyte activity’, ‘cytokine activity’, and ‘positive regulation of protein phosphorylation’. ( E ) Heatmap comparing normalized transcript counts (blue – low to red – high) of genes exclusively upregulated in the lean group following E. coli infection. ( F ) Experimental design for measuring ex vivo phagocytosis by cord blood monocytes. ( G ) Bar graph depicting colorimetric readout of phagocytosed E. coli particles by UCB monocyte. * or /#- p<0.05, #### - p<0.0001.

Article Snippet: Approximately 5x10 5 MACS purified UCB monocytes were cultured with RSV (Human respiratory syncytial virus ATCC VR-1540, Manassas, VA) or E. coli ( Escherichia coli (Migula) Castellani and Chalmers ATCC 11775, Manassas, VA) or left untreated in RP10 medium for 16 hr at 37 °C.

Techniques: In Vitro, Purification, Cell Culture, RNA Sequencing Assay, Luminex, Infection, Transformation Assay, Functional Assay, Cell Differentiation, Activity Assay, Ex Vivo

Journal: eLife

Article Title: Maternal obesity blunts antimicrobial responses in fetal monocytes

doi: 10.7554/eLife.81320

Figure Lengend Snippet: ( A ) Bar graph representing the number of upregulated (red) or downregulated (blue) differentially expressed genes (DEG) in each group. ( B ) Venn diagram representing the number of downregulated DEG following E. coli stimulation in each group. ( C ) Functional enrichment of downregulated DEG detected in both groups (top, green), or exclusively in the lean (middle, blue) and obese (bottom, orange) groups. ( D ) Violin plots comparing fold changes of DEGs downregulated following E. coli stimulation and involved in antigen presentation.

Article Snippet: Approximately 5x10 5 MACS purified UCB monocytes were cultured with RSV (Human respiratory syncytial virus ATCC VR-1540, Manassas, VA) or E. coli ( Escherichia coli (Migula) Castellani and Chalmers ATCC 11775, Manassas, VA) or left untreated in RP10 medium for 16 hr at 37 °C.

Techniques: Functional Assay

Journal: eLife

Article Title: Maternal obesity blunts antimicrobial responses in fetal monocytes

doi: 10.7554/eLife.81320

Figure Lengend Snippet: ( A ) Bar graph representing the number of upregulated (red) or downregulated (blue) DEG in each group. ( B ) Heatmap of upregulated genes involved in interferon signaling pathway. ( C ) Surface protein levels of interferon receptor 1 (IFNAR1) represented as the fraction of positive cells (top) or median fluorescent intensity (bottom). *- p<0.05, ***- p<0.001. ( D ) Venn diagram representing the number of downregulated DEG following RSV stimulation. ( E ) Functional enrichment of downregulated DEG detected in both groups (top, green), or exclusively in the lean group (top, blue) and obese group (bottom, orange). ( F ) Four-way venn comparing overall gene expression (both up- and down-regulated genes) changes following E. coli and RSV infections. ( G ) Five way functional enrichment of unique DEG and the 79 DEG induced in all groups induced following in vitro infection with E. coli and RSV.

Article Snippet: Approximately 5x10 5 MACS purified UCB monocytes were cultured with RSV (Human respiratory syncytial virus ATCC VR-1540, Manassas, VA) or E. coli ( Escherichia coli (Migula) Castellani and Chalmers ATCC 11775, Manassas, VA) or left untreated in RP10 medium for 16 hr at 37 °C.

Techniques: Functional Assay, Expressing, In Vitro, Infection

Journal: eLife

Article Title: Maternal obesity blunts antimicrobial responses in fetal monocytes

doi: 10.7554/eLife.81320

Figure Lengend Snippet: ( A ) Dot plots comparing median fluorescence intensity (MFI) ± SEM of phosphorylated signaling molecules downstream of TLR4 sensing (n=5/group). *- p<0.05, **- p<0.01. ( B ) Representative brightfield and fluorescent images of stimulated and unstimulated UCB monocytes profiled using imagine flow cytometry (n=3–4/group). NF-kB (p50–AF488) and nucleus (7-AAD) are shown in green and red respectively. Surface stains for CD14 and HLA-DR are shown in aqua and fuchsia respectively. Overlay of NF-kB and nuclei stain was used to determine translocation within CD14 + HLA-DR+monocytes. Bar graph comparing percentage translocated cells following LPS stimulation in lean and obese groups. ( C ) Bar graph comparing changes in trimethyl modification on H3K9 residues following LPS stimulation (relative to no stimulation controls) detected using flow cytometry. ( D ) Graph representing the kinetics of ECAR of stimulated monocytes following glucose injection and blockade of glycolysis. ( E ) Bar graph comparing DAR frequencies in each group following LPS stimulation. ( F ) Heatmap demonstrating overall accessibility differences following LPS stimulation around the promoter. ( G ) Over-represented transcription factors identified from motif analysis of DARs more open in lean compared to obese groups following LPS stimulation. X-axis represented percentage of peaks with motifs identified and color represents p-value on log10 scale ( H ) Pileups of key inflammatory loci post LPS stimulation ( I ) Four-way Venn of genes accessible (ATAC-Seq) exclusively in the lean group post LPS stimulation with genes exclusively upregulated (RNA-Seq) in the lean group following LPS, E. coli and RSV infection. Select overlapping genes are highlighted. ( J ) UMAP of single nuclei ATAC-Seq of LPS stimulated and sorted monocytes. ( K ) Feature plots demonstrating a cluster of activated monocytes (cluster 1). Color intensity represents fragments mapping to open chromatin regions. ( L ) Proportions of monocytes within each group mapping to activated monocyte cluster. ( M ) Pileups of inflammatory loci in activated monocytes with/without LPS stimulation. * - p<0.05; ** - p<0.01.

Article Snippet: Approximately 5x10 5 MACS purified UCB monocytes were cultured with RSV (Human respiratory syncytial virus ATCC VR-1540, Manassas, VA) or E. coli ( Escherichia coli (Migula) Castellani and Chalmers ATCC 11775, Manassas, VA) or left untreated in RP10 medium for 16 hr at 37 °C.

Techniques: Fluorescence, Flow Cytometry, Staining, Translocation Assay, Modification, Injection, RNA Sequencing Assay, Infection

Journal: eLife

Article Title: Maternal obesity blunts antimicrobial responses in fetal monocytes

doi: 10.7554/eLife.81320

Figure Lengend Snippet: ( A ) NHP model of maternal diet-induced obesity. Fetal (GD130) PBMC, ileal leukocytes, and spleenocytes were obtained from rhesus macaque born to dams on a control chow diet (n=3) and western-style diet (n=4). Fetal macrophages from ileal lamina propria lymphocytes and splenocytes were FACS-sorted and cultured overnight with E. coli . Supernatants were collected and cytokines/chemokines were measured using Luminex. ( B ) Bar graph representing the frequency (mean and ± SEM) of responding fetal monocytes to LPS in PBMC isolated from GD130 fetal circulation measured by intracellular cytokine staining. ( C ) Frequencies of CD14 + HLA-DR+macrophages within live total ileal leukocytes (top) or live CD3-CD20- leukocytes (bottom) ( D–E ) Median fluorescence intensity (MFI) of ( D ) M1-associated HLA-DR and ( E ) M2-associated CD209 and CD206 within live macrophage population in ileum (top) and spleen (bottom). ( F–G ) Bubble plots comparing significantly different cytokine/chemokine levels in response to E. coli stimulation in ( F ) gut (ileum) and ( G ) splenic macrophages. Statistical differences between chow and WSD groups are highlighted. *-p<0.05.

Article Snippet: Approximately 5x10 5 MACS purified UCB monocytes were cultured with RSV (Human respiratory syncytial virus ATCC VR-1540, Manassas, VA) or E. coli ( Escherichia coli (Migula) Castellani and Chalmers ATCC 11775, Manassas, VA) or left untreated in RP10 medium for 16 hr at 37 °C.

Techniques: Western Blot, Cell Culture, Luminex, Isolation, Staining, Fluorescence