Journal: Scientific Reports

Article Title: Selective right heart valve remodelling in a mouse model of carcinoid disease revealed by high-resolution episcopic microscopy

doi: 10.1038/s41598-025-16064-8

Figure Lengend Snippet: Validation of B16F0-Tph1 cell line. ( a ) Representative fluorescence microscopy image showing GFP expression in B16F0-Tph1 cells. ( b ) Flow cytometry analysis demonstrating 85% GFP-positive cells at passage 2 (P2). Red histogram: non-modified B16F0 cells; green histogram: B16F0-Tph1 cells. ( c ) 5-HT concentration in culture supernatants showing sustained production in B16F0-Tph1 cells across passages (P1-P5) compared to B16F0 controls. Values represent median ± IQR from n = 3 independent experiments per condition. Statistical significance was determined using Kolmogorov–Smirnov test (* p < 0.05).

Article Snippet: B16F0 mouse melanoma cells from American Type Culture Collection (ATCC, USA) were genetically modified to produce 5-HT.

Techniques: Biomarker Discovery, Fluorescence, Microscopy, Expressing, Flow Cytometry, Modification, Concentration Assay

Journal: Scientific Reports

Article Title: Selective right heart valve remodelling in a mouse model of carcinoid disease revealed by high-resolution episcopic microscopy

doi: 10.1038/s41598-025-16064-8

Figure Lengend Snippet: Macroscopic dissemination of tumor cells over time and blood 5-HT elevation in RV following intrahepatic B16F0-Tph1 implantation. ( a ) Representative images of mice injected with 1000 B16F0-Tph1 cells in the liver, showing progression of tumor spread at 5, 6 and 7 weeks post-injection. White arrows indicate extrahepatic dissemination (peritoneal carcinomatosis at 6 weeks and mediastinal lymph node metastases at 7 weeks). No extrahepatic dissemination was detected at 5 weeks, establishing this timepoint as optimal for the main study. ( b ) Blood 5-HT concentrations measured in the right ventricle (RV) and systemic circulation 5 weeks after intrahepatic implantation in B16F0-Tph1 and B16F0 mice. Data are presented as individual points with median ± IQR; n = 3 per group. Statistical significance was determined using Kolmogorov–Smirnov test (* p < 0.05 and ** p < 0.01).

Article Snippet: B16F0 mouse melanoma cells from American Type Culture Collection (ATCC, USA) were genetically modified to produce 5-HT.

Techniques: Injection

Journal: Scientific Reports

Article Title: Selective right heart valve remodelling in a mouse model of carcinoid disease revealed by high-resolution episcopic microscopy

doi: 10.1038/s41598-025-16064-8

Figure Lengend Snippet: Analysis of urinary 5-HT and metabolite. ( a ) Log-transformed urinary serotonin (5-HT/creatinine) levels in sham (n = 7) and B16F0-Tph1 (n = 7) mice at 5 weeks post-surgery. ( b ) Log-transformed urinary 5-hydroxyindoleacetic acid (5-HIAA/creatinine) levels in the same groups. Data are shown as individual points with median ± IQR. Statistical significance was determined using Kolmogorov–Smirnov test (**** p < 0.0001, ns: not significant).

Article Snippet: B16F0 mouse melanoma cells from American Type Culture Collection (ATCC, USA) were genetically modified to produce 5-HT.

Techniques: Transformation Assay

Journal: Scientific Reports

Article Title: Selective right heart valve remodelling in a mouse model of carcinoid disease revealed by high-resolution episcopic microscopy

doi: 10.1038/s41598-025-16064-8

Figure Lengend Snippet: Heterogeneous signal intensity distribution in pulmonary and tricuspid valves of B16F0-Tph1 mice. ( a ) Representative HREM images with grayscale intensity overlays (blue: low intensity; yellow: high intensity). Sham valves show uniform low-intensity signal, while B16F0-Tph1 valves exhibit spatially heterogeneous intensity patterns, with distinct high-intensity regions adjacent to low-intensity zones. ( b ) Normalized intensity histograms demonstrate a rightward shift in B16F0-Tph1 mice, with new high-intensity voxel populations (120–180 range) absent in sham controls. Scale bar: 0.5 mm.

Article Snippet: B16F0 mouse melanoma cells from American Type Culture Collection (ATCC, USA) were genetically modified to produce 5-HT.

Techniques:

Journal: Scientific Reports

Article Title: Selective right heart valve remodelling in a mouse model of carcinoid disease revealed by high-resolution episcopic microscopy

doi: 10.1038/s41598-025-16064-8

Figure Lengend Snippet: High-resolution episcopic microscopy (HREM) analysis reveals carcinoid-induced pathology in the tricuspid valve subvalvular apparatus of B16F0-Tph1 mice. Representative HREM images of the tricuspid valve (white arrow) and associated chordae tendineae (white asterisk) in Sham ( a ) and B16F0-Tph1 ( b ) mice. Quantitative analysis shows increased chordae tendineae length ( c ) and thickness ( d ) in B16F0-Tph1 compared to Sham mice. Data are shown as individual points with median ± IQR; n = 7 per group. Statistical significance was determined using Kolmogorov–Smirnov test (*** p < 0.0001, * p < 0.05, ns: not significant). Scale bar: 0.5 mm.

Article Snippet: B16F0 mouse melanoma cells from American Type Culture Collection (ATCC, USA) were genetically modified to produce 5-HT.

Techniques: Microscopy

Journal: Scientific Reports

Article Title: Selective right heart valve remodelling in a mouse model of carcinoid disease revealed by high-resolution episcopic microscopy

doi: 10.1038/s41598-025-16064-8

Figure Lengend Snippet: Cardiac valve thickness patterns. Valve thickness was measured at 6 equidistant points from free edge (position 1) to base (position 6) in B16F0-Tph1-treated mice (red lines) and sham controls (blue lines) in all cusps and leaflets (triangle = anterior, circle = posterior or left, inverted triangle = septal or right). Solid lines are spline curves of actual data (dotted lines). Data are presented as median ± IQR; n = 7 per group.

Article Snippet: B16F0 mouse melanoma cells from American Type Culture Collection (ATCC, USA) were genetically modified to produce 5-HT.

Techniques:

Journal: Scientific Reports

Article Title: Selective right heart valve remodelling in a mouse model of carcinoid disease revealed by high-resolution episcopic microscopy

doi: 10.1038/s41598-025-16064-8

Figure Lengend Snippet: Valvular thickness and length analysis. Comparison between B16F0-Tph1 mice (red dots) and sham controls (blue dots) across different valve leaflets and cusps. Given the absence of position effect (Fig. ), the six measurements along each leaflet/cusp were used as repeated measures. Tricuspid valve showed significant thickening across all cusps, while pulmonary valve demonstrated significant thickening in anterior and left cusps but not the right cusp. Both tricuspid and pulmonary valves showed decreased length in anterior cusps, with additional reductions in posterior (tricuspid) and left (pulmonary) cusps. Data are presented as median ± IQR; n = 7 per group. Statistical significance was determined using Kolmogorov–Smirnov test (*** p < 0.0001, ** p < 0.0001, * p < 0.0001, ns: not significant).

Article Snippet: B16F0 mouse melanoma cells from American Type Culture Collection (ATCC, USA) were genetically modified to produce 5-HT.

Techniques: Comparison

Journal: Scientific Reports

Article Title: Selective right heart valve remodelling in a mouse model of carcinoid disease revealed by high-resolution episcopic microscopy

doi: 10.1038/s41598-025-16064-8

Figure Lengend Snippet: Volumetric analysis and correlation of cardiac valves with urinary 5-HT. ( a ) 3D rendering examples of measured volumes for pulmonary and tricuspid valves in sham (blue) and B16F0-Tph1 (red) mice. ( b ) Quantitative comparison of valve volumes showing significantly increased volumes in B16F0-Tph1 mice (n = 7) compared to sham (n = 7) and B16F0 controls (n = 2) for both pulmonary and tricuspid valves. Data are presented as median ± IQR. ( c ) Linear regression analysis showing the relationship between urinary log(5-HT/creatinine) and valve volume (mm 3 ) in tricuspid valve (left) and pulmonary valve (right). Blue dots represent sham animals and red dots represent B16F0-Tph1 animals. Solid lines indicate linear regression with 95% confidence intervals (dashed lines). Dotted vertical and horizontal lines represent threshold values calculated using stepwise regression, yielding high diagnostic performance (pulmonary valve: 86% sensitivity and specificity; tricuspid valve: 100% sensitivity and specificity) for detecting pathological valve thickening. Values are presented as median ± IQR. Scale bar: 0.5 mm. Statistical significance was determined using one-way ANOVA followed by Tukey’s multiple comparisons test (*** p < 0.001; ns, not significant).

Article Snippet: B16F0 mouse melanoma cells from American Type Culture Collection (ATCC, USA) were genetically modified to produce 5-HT.

Techniques: Comparison, Diagnostic Assay

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Pharmacological attenuation of melanoma by tryptanthrin pertains to the suppression of MITF-M through MEK/ERK signaling axis

doi: 10.1007/s00018-022-04476-y

Figure Lengend Snippet: Tpn induces melanoma cell apoptosis A A panel of human cancer cell lines of different tissue origin was treated with different concentrations of Tpn. After 72 h, the cell viability was assessed by MTT assay. B Human melanoma cell lines, A375, SK- MEL-28, SK-MEL-2, and the murine melanoma cell line, B16F10 were compared with normal immortalized melanocytes, HEMa-LP for assessing the cell viability on treatment with increasing concentrations of Tpn. C Tpn, at its IC50 concentration, was evaluated for the induction of early and late stages of apoptosis using Annexin V-FITC-PI-FACS analysis in A375 cells. D DAPI staining to evaluate nuclear condensation induced by Tpn, post 6 h treatment, in A375 cells, 60 X magnifications is shown in the inset. E Immunoblot for caspase activation and PARP cleavage indicating the apoptotic mode of cell death induced by Tpn at 48 h incubation in A375 cells. F To evaluate the mechanism of apoptosis induced upon Tpn treatment immunoblot was performed to check the expression of tumor suppressor p53, pro-apoptotic proteins BAX and Bid and anti-apoptotic proteins Bcl-2 and Bcl-xL. β-actin was used as a loading control. G Cellular ROS levels analyzed upon Tpn treatment in A375 cells by H2DCF-DA assay. H A375 Mito roGFP ratiometric probe showing increased redox ratio after 48 h of Tpn treatment, which indicates increased mitochondrial ROS levels

Article Snippet: Cell lines and compound The cell lines A375 and B16F10 were procured from National Centre for Cell Sciences (Pune, India).

Techniques: MTT Assay, Concentration Assay, Staining, Western Blot, Activation Assay, Incubation, Expressing, Control

Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Pharmacological attenuation of melanoma by tryptanthrin pertains to the suppression of MITF-M through MEK/ERK signaling axis

doi: 10.1007/s00018-022-04476-y

Figure Lengend Snippet: Suppression of melanoma migration and metastasis by Tpn A Scratch wound assay was performed to check the ability of Tpn to prevent the migratory property of A375 cells. Graphical representation of the wound area (Pixels) of n = 3 experiments. B Immunoblotting was carried out to check the cytoplasmic and nuclear levels of β-catenin. Vinculin was used as loading control for cytoplasmic fraction and Lamin B was used as nuclear control. C TOP-FOP-dual-luciferase assay to check the activation of β-catenin upon Tpn treatment. LiCl was used as a positive control. D Black melanotic nodules were observed in the lungs of control groups of the B16F10 tail vein metastasis model. Tpn treatment group showed less number of melanotic nodules. E Graphical representation showing statistically significant (p-value < 0.0001) reduction in the number of metastatic nodules upon Tpn treatment in B16F10 tail vein models. F Colorless foci were observed in the lungs of control groups of A375 tail vein metastasis model. Tpn treatment group showed less number of lung nodules. G Graphical representation showing statistically significant (p-value < 0.0097) reduction in the number of metastatic nodules upon Tpn treatment in A375-Luc+ tail vein models. H Ex vivo images of the organs of A375-Luc+ tail vein metastasis model, representative animals of control and Tpn treatment are shown (n = 3). ROI values indicate the signal emitted (photons). I Luciferase activity was performed using the Dual-luciferase assay kit, showing the relative luciferase signals of the organs of the control and treatment group of three technical replicates (mean ± SD). J Histopathological verification of the hepatic tissues was done by H and E staining. K Chromatogram showing the peak response of internal standard (2-Hydroxy acetophenone) and Tpn. L The serum retention of different concentrations of liposomal Tpn was analyzed by HPLC

Article Snippet: Cell lines and compound The cell lines A375 and B16F10 were procured from National Centre for Cell Sciences (Pune, India).

Techniques: Migration, Scratch Wound Assay Assay, Western Blot, Control, Luciferase, Activation Assay, Positive Control, Ex Vivo, Activity Assay, Staining