Journal: Scientific Reports

Article Title: Psoriatic disease is associated with systemic inflammation, endothelial activation, and altered haemostatic function

doi: 10.1038/s41598-021-90684-8

Figure Lengend Snippet: Box-and-whisker plots illustrating the distribution of parameters assessed in this study in healthy individuals and psoriatic patients. ( A – D ) indicates V-PLEX panel results, ( E ) indicates sSELP ELISA results, ( F – L ) indicates TEG results, and ( M – O ) indicates fibrin fibre diameter results. Asterisks indicate statistically significant differences between the groups, as determined by either an unpaired t-test or Mann–Whitney test, where **** p < 0.0001, *** p < 0.001, ** p < 0.01, and * p < 0.05. Box-and-whisker plots were produced using GraphPad Prism version 8.4.3.

Article Snippet: Box-and-whisker plots were produced using GraphPad Prism version 8.4.3.

Techniques: Whisker Assay, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY, Produced

Journal: Scientific Reports

Article Title: Psoriatic disease is associated with systemic inflammation, endothelial activation, and altered haemostatic function

doi: 10.1038/s41598-021-90684-8

Figure Lengend Snippet: Frequency bar graphs of fibrin fibre diameter distribution. ( A ) indicates healthy individuals and ( B ) indicates psoriatic patients. Bar graphs were produced using GraphPad Prism version 8.4.3.

Article Snippet: Box-and-whisker plots were produced using GraphPad Prism version 8.4.3.

Techniques: Produced

Journal: Scientific Reports

Article Title: Deprogramming metabolism in pancreatic cancer with a bi-functional GPR55 inhibitor and biased β 2 adrenergic agonist

doi: 10.1038/s41598-022-07600-x

Figure Lengend Snippet: ( R , S ′)-MNF inhibits proliferation of PDAC cell lines. ( A ) ( R , S ′)-MNF (structure depicted) treatment reduces PANC-1 cell proliferation. [ 3 H]-Thymidine incorporation into PANC-1 cells was assessed after incubation with increasing concentrations of ( R , S ′)-MNF for 24 h. Data are expressed as mean ± SD from 3 independent experiments. The calculated IC 50 value was 0.11 ± 0.08 µM. (B) PDAC cell lines were exposed to ( R , S ′)-MNF for 72 h. Next, the viability of the cells was assessed using the Sulforhodamine B (SRB) assay. ( R , S ′)-MNF inhibited the survival of the MIA PaCa-2, PSN-1, PANC-1, HPAC, and Capan-1 cells with the IC 50 of 5.2, 8.3, 9.6, 5.0, and 6.8 µM, respectively. (C) Upper panel , Immunoblot showing levels of phosphoactive and total forms of ERK after a 20-min pretreatment of serum-depleted PANC-1 cells with or without 1 µM ( R , S ′)-MNF followed by short term incubation with the GPR55 agonist O-1602 (10 µM, 20 min). Lower panel , Protein bands were quantified by densitometry, and the ratio of phosphorylated/total forms of ERK1/2 was calculated and plotted relative to O-1602-treated cells. Values are represented as boxplots with n = 6. Data analysis: one-way ANOVA followed by Tukey’s post-hoc test; **, *** P < 0.01, 0.001 vs. control or vs. marked treatments. All graphs were generated with GraphPad Prism v.8.4.3 (GraphPad Software, Inc., La Jolla, CA).

Article Snippet: The dose- and time-dependent trajectories as well as the box plot were generated with GraphPad Prism v.8.4.3 (GraphPad Software, Inc., La Jolla, CA).

Techniques: Incubation, Sulforhodamine B Assay, Western Blot, Generated, Software

Journal: Scientific Reports

Article Title: Deprogramming metabolism in pancreatic cancer with a bi-functional GPR55 inhibitor and biased β 2 adrenergic agonist

doi: 10.1038/s41598-022-07600-x

Figure Lengend Snippet: Biological activity of ( R , S ′)-MNF in PANC-1 carcinoma cell line. ( A ) PANC-1 cells were treated with 5, 10, 20 µM ( R , R ′)-MNF (blue), ( R , S ′)-MNF (red) or with vehicle (0.1% DMSO; black) for 20 min and lysed. Level of phosphorylated and total ERK, AKT and eEF2 was assessed by immunoblotting in the obtained lysates. Values are represented as boxplots with n = 6. ( B ) Immunoblots showing levels of phosphorylated and total forms of ERK, AKT and eEF2 after a 20-min pretreatment of serum-depleted PANC-1 cells with or without 50 nM ICI followed by incubation either with ( R , R ′)-MNF or ( R , S ′)-MNF (1 µM, 20 min). ( C ) Protein bands were quantified by densitometry, and the ratio of phosphorylated/total forms of the proteins was calculated and plotted relative to vehicle-treated cells. Values are represented as boxplots with n = 7. ( D ) PANC-1 cells were treated as on panel F and phosphorylated targets of PKA were visualized as a marker for PKA activation. ( E ) Effect of ( R , R ′)-MNF or ( R , S ′)-MNF on the PKI-dependent (10 µM) activation of ERK and AKT. Representative blots are depicted on panel ( F ). ( G ) β 2 -AR activation inhibits ERK and AKT phosphorylation. Illustration was created using Canvas X Draw v.7.0.2 for macOS (Canvas GFX, Boston, MA; canvasgfx.com ). Data analysis: one-way ANOVA followed by Tukey’s post-hoc test; *, **, ***, **** P < 0.05, 0.01, 0.001, 0.0001 vs. control or vs. marked treatments. All box plots were generated with GraphPad Prism v.8.4.3 (GraphPad Software, Inc., La Jolla, CA).

Article Snippet: The dose- and time-dependent trajectories as well as the box plot were generated with GraphPad Prism v.8.4.3 (GraphPad Software, Inc., La Jolla, CA).

Techniques: Activity Assay, Western Blot, Incubation, Marker, Activation Assay, Generated, Software

Journal: Scientific Reports

Article Title: Deprogramming metabolism in pancreatic cancer with a bi-functional GPR55 inhibitor and biased β 2 adrenergic agonist

doi: 10.1038/s41598-022-07600-x

Figure Lengend Snippet: ( R , S ′)-MNF treatment reduces PANC-1 tumor growth in a mouse xenograft model. ( A ) Protocol design. ( B ) Tumor volume was determined after i.p. administration of vehicle (1% hydroxypropyl-β-cyclodextrin), 20 mg kg −1 (Arm 1) or 40 mg kg −1 ( R , S ′)-MNF (Arm 2) once daily for 5 days per week for 3 weeks (n = 10 per group). The black arrow depicts the last day of ( R , S ′)-MNF administration. Data represent mean + SD. ** P < 0.01 vs. control group of mice. Representative images of mice ( C ) and excised tumors ( D ) are shown. ( E ) Plasma lactate levels were measured on Day 8 (Pretreatment, open symbols) and at completion of the ( R , S ′)-MNF treatment (After treatment, filled symbols) (n = 8–10 per group). * P < 0.05, *** P < 0.001; # P < 0.05 vs. control group of mice at the completion of the study. The dose- and time-dependent trajectories as well as the box plot were generated with GraphPad Prism v.8.4.3 (GraphPad Software, Inc., La Jolla, CA).

Article Snippet: The dose- and time-dependent trajectories as well as the box plot were generated with GraphPad Prism v.8.4.3 (GraphPad Software, Inc., La Jolla, CA).

Techniques: Generated, Software

Journal: Scientific Reports

Article Title: Deprogramming metabolism in pancreatic cancer with a bi-functional GPR55 inhibitor and biased β 2 adrenergic agonist

doi: 10.1038/s41598-022-07600-x

Figure Lengend Snippet: ( R , S ′)-MNF treatment elicits distinct metabolomics signature in PANC-1 tumor xenografts. ( A ) Heatmap depicting the P -values [− log10( P )] from the univariate analyses of the 35 metabolites shown to be significantly altered in response to ( R,S ′)-MNF. Purple arrows indicate l -carnitine and different species of acylcarnitines. The heatmap was created using Microsoft Excel v.16.56 for Mac (Redmond, WA). ( B , C ) The Over Representation Analysis (ORA) module of MetaboAnalyst 5.0 was used to provide better representation of the sub-classes of metabolites ( B ) and lipids ( C ) with their enrichment significance in our dataset of 35 significantly altered compounds. Significantly enriched ‘fatty acyl carnitines’ are indicated with purple arrow. ( D ) Numbers of biochemicals related to lipid metabolism that were differentially expressed in the PANC-1 tumor xenografts from ( R , S ′)-MNF vs. control groups; n = 9–10 mice per group. ( E ) Relative abundance of selected biochemicals between the two experimental groups, as quantified by targeted metabolomics. See Supporting Information Table for complete list. *, **, **** P < 0.05, 0.01, 0.0001 vs. vehicle controls. The box plots were generated with GraphPad Prism v.8.4.3 (GraphPad Software, Inc., La Jolla, CA). ( F ) Altered cysteine/methionine metabolism in PANC-1 tumor xenografts through formation of ophthalmic acid upon ( R , S ′)-MNF treatment. ( G ) Schematic representation of the change in metabolites associated with pyrimidine synthetic and salvage pathways following ( R , S ′)-MNF treatment. Metabolic pathway schemes were drawn using Canvas X Draw v.7.0.2 for macOS (Canvas GFX, Boston, MA; canvasgfx.com ). DG, DAG, diacylglycerol; TG, TAG, triacylglycerol; PE, phosphatidylethanolamine; PC, phosphatidylcholine; PS, phosphatidylserine.

Article Snippet: The dose- and time-dependent trajectories as well as the box plot were generated with GraphPad Prism v.8.4.3 (GraphPad Software, Inc., La Jolla, CA).

Techniques: Generated, Software

Journal: Scientific Reports

Article Title: Deprogramming metabolism in pancreatic cancer with a bi-functional GPR55 inhibitor and biased β 2 adrenergic agonist

doi: 10.1038/s41598-022-07600-x

Figure Lengend Snippet: ( R , S ′)-MNF treatment elicits distinct gene expression signature in PANC-1 tumor xenografts. ( A ) Percent of genes and GO Terms that were up- and down-regulated in the ( R , S ′)-MNF:vehicle pairwise comparison. ( B ) Enrichment of top GO Terms that were up- (positive Z-scores) and down-regulated (negative Z-scores) after ( R , S ′)-MNF treatment. ( C ) Heatmap depicting the expression of genes implicated in fatty acid oxidation that were significantly impacted by ( R , S ′)-MNF. ( D ) Enrichment of top GO Terms related to Mitochondrial Pathway dataset from the ( R , S ′)-MNF:vehicle pairwise comparison. The complete list can be found in Supporting Information Table . ( E ) Heatmap showing the statistically significant Z-ratio changes in expression of selected genes in the ( R , S ′)-MNF:vehicle pairwise comparison. These genes are implicated in a handful of pro-oncogenic signaling pathways, including Hippo, Wnt-β-catenin, and EGF receptor. ( F ) Input for the multi-omics Joint Pathway Analysis (JPA) from MetaboAnalyst 5.0 consisted of the 74 metabolites gathered in Supporting Information Table and the 1890 differentially expressed transcripts identified by microarray profiling. y axis, enrichment significance; x axis, pathway impact for network topology. Dotted line highlights pathways significantly impacted as defined by enrichment significance P < 0.05 [log( P ) > 1.3]. ( G ) String protein–protein interaction database (v 11.5; string-db.org ) was used to visualize the putative impact of ( R,S ′)-MNF on functional c-Myc (left panel) and HIF-1α (right panel) protein–protein association based on the transcriptional responses of PANC-1 tumors to ( R,S ′)-MNF treatment. Red nodes, significantly upregulated genes; blue nodes, significantly downregulated genes. In String network analysis, nodes represent proteins whereas edges represent functional protein–protein associations. If the number of edges for analyzed dataset is higher than expected for a random set of proteins (the ‘exp.’ value), the clustered proteins are biologically connected as a group. The average node degree indicates how many interactions a protein has on average in the network. The clustering coefficient is a measure of how connected the nodes in the network are. Protein–protein interaction (PPI) enrichment P -value is a measure of significance for biological association of investigated proteins. ( H ) Lysates prepared from PANC-1 tumor xenografts were separated by SDS-PAGE and immunoblotted using primary antibodies specific for YAP, HIF-1α, c-Myc, β-catenin, CYR61, cyclin D1, and β-actin (n = 9–10 mice per group). Representative blots are depicted. ( I ) Scatter plots depict densitometric quantitation of each protein band followed by normalization to β-actin levels. *, ** P < 0.05, < 0.01 vs. control. The heatmaps were created using Microsoft Excel v.16.56 for Mac (Redmond, WA). The bar graphs and scatter plots were generated with GraphPad Prism v.8.4.3 (GraphPad Software, Inc., La Jolla, CA).

Article Snippet: The dose- and time-dependent trajectories as well as the box plot were generated with GraphPad Prism v.8.4.3 (GraphPad Software, Inc., La Jolla, CA).

Techniques: Expressing, Microarray, Functional Assay, SDS Page, Quantitation Assay, Generated, Software