Journal: Translational Oncology

Article Title: Improving photodynamic therapy efficacy in bladder cancer using polymer micelle-encapsulated pheophorbide a

doi: 10.1016/j.tranon.2026.102687

Figure Lengend Snippet: Encapsulation of pheophorbide a enhances PDT efficacy in 3D tumor spheroids. A. Scanning electron microscopy (SEM) images showing the surface morphology of T24 and SW780 spheroids. B. General appearance of bladder tumor spheroids before treatment. Red arrows indicate detached cyst-like vesicles, i.e. “microbladders” (luminal cavities characteristic of urothelial differentiation); dotted circles highlight internal cyst-like structures embedded within the spheroids. C. Counterstained semi-thin transverse sections (500 nm) of spheroids revealing internal cyst-like structures (dotted circles). D. Two-photon microscopy imaging of pheophorbide a (Pheo) (1 µM) penetration in T24 spheroids after 30 min incubation at 37 °C either in its free or encapsulated formulation. Poly (ethylene oxide)-block-poly (ε-caprolactone) (PEO 5000 -PCL 4000 ) empty micelles; Pheophorbide encapsulated in PEO-PCL micelles (Pheo-PEOPCL). Cyan: nuclei (Hoechst); red: pheophorbide fluorescence. E. Spheroid viability assessed by intracellular ATP quantification at 3- and 6-days post-PDT ([Pheo] = 3 µM). Results include data from 1 to 6 independent experiments (N), with a cumulative number of biological replicates (n) ranging from 6 to 39.

Article Snippet: Pheophorbide a (Pheo) was purchased from Frontier Scientific (Logan, Utah, USA).

Techniques: Encapsulation, Electron Microscopy, Microscopy, Imaging, Incubation, Formulation, Blocking Assay, Fluorescence

Journal: Translational Oncology

Article Title: Improving photodynamic therapy efficacy in bladder cancer using polymer micelle-encapsulated pheophorbide a

doi: 10.1016/j.tranon.2026.102687

Figure Lengend Snippet: Encapsulation of pheophorbide a in PEO-PCL micelles enhances PDT-induced cytotoxicity in 2D high-grade (T24) and low-grade (SW780) bladder cancer cell cultures. A. Cells were treated with empty poly (ethylene oxide)-block-poly (ε-caprolactone) PEO-PCL micelles (PEOPCL) at polymer concentrations equivalent to those used in pheo-loaded micelles, based on the 1:30 drug-to-polymer weight ratio. B. Cells were exposed to increasing concentrations of free pheophorbide a (Pheo) (nM). C. Cells were treated with increasing concentrations of Pheo encapsulated in PEO-PCL micelles (pheo–PEOPCL) (nM). D. Half-maximal inhibitory concentration (IC₅₀) determination for the Pheo–PEOPCL condition. Cell confluence was monitored by videomicroscopy for 72 h post-PDT ( N > 1; n = 6).

Article Snippet: Pheophorbide a (Pheo) was purchased from Frontier Scientific (Logan, Utah, USA).

Techniques: Encapsulation, Blocking Assay, Polymer, Concentration Assay

Journal: Translational Oncology

Article Title: Improving photodynamic therapy efficacy in bladder cancer using polymer micelle-encapsulated pheophorbide a

doi: 10.1016/j.tranon.2026.102687

Figure Lengend Snippet: Reduced wound closure in bladder cancer cell monolayers following photodynamic treatment with encapsulated pheophorbide a . A. Kinetics of wound closure over 24 h post-PDT in T24 (grade 3) and SW780 (grade 1) monolayers, monitored by videomicroscopy. Cells were treated with increasing concentrations of pheophorbide a encapsulated in poly (ethylene oxide)-block-poly (ε-caprolactone) PEO-PCL micelles (Pheo-PEO-PCL) (nM). B. Quantification of wound closure ( %) at 24 h post-PDT across the different Pheo-PEO-PCL concentrations. Relative wound density ( %) reflects the proportion of the scratch area repopulated by migrating cells. Data are expressed as mean ± SEM. Statistical analysis was performed using one-way ANOVA followed by Dunnett’s multiple comparisons test versus control (0 nM).

Article Snippet: Pheophorbide a (Pheo) was purchased from Frontier Scientific (Logan, Utah, USA).

Techniques: Blocking Assay, Control

Journal: Translational Oncology

Article Title: Improving photodynamic therapy efficacy in bladder cancer using polymer micelle-encapsulated pheophorbide a

doi: 10.1016/j.tranon.2026.102687

Figure Lengend Snippet: Proof of concept of photodynamic therapy efficacy in a complex 3D engineered bladder tumor model. A. Histological cross-sections of the vesical reconstructed tissues stained with Masson’s trichrome, 72 hours after PDT with 3 µM of either free pheophorbide a (Pheo) or pheophorbide a encapsulated in poly (ethylene oxide)-block-poly (ε-caprolactone) PEO-PCL micelles (Pheo–PEO-PCL). Cells appear in red, and stromal collagens in blue. Tumor cells are outlined with black dotted lines. Representative images are shown. B. Tissue viability assessed using the PrestoBlue assay at 48- and 72-hours post-PDT on engineered bladder substitutes implanted with T24 or SW780 spheroids. Results are expressed as a percentage of the untreated control at each timepoint ± SEM.

Article Snippet: Pheophorbide a (Pheo) was purchased from Frontier Scientific (Logan, Utah, USA).

Techniques: Staining, Blocking Assay, Prestoblue Assay, Control

Journal: Human Genetics and Genomics Advances

Article Title: Aberrant N-glycosylation may be a therapeutic target in carriers of a common and highly pleiotropic variant in the manganese transporter ZIP8

doi: 10.1016/j.xhgg.2025.100517

Figure Lengend Snippet: The ileal epithelium is dominated by L-PHA + , complex N-glycans in healthy subjects and by truncated, sWGA + N-glycans in active Crohn disease (A) Schematic of N-glycan branching cascade demonstrating the glycan targets of lectins and manganese (Mn) utilization. sWGA (green) stains terminal GlcNAc residues. L-PHA (red) stains complex tri- and tetra-antennary N-glycans. Relative UDP-GlcNAc affinity of glycosyltransferases is also represented (300-fold change from Mgat1 to Mgat5). Figure generated in Biorender, informed by Cummings et al. (ref. ). (B) Confocal laser-scanning triple-label immunofluorescence microscopy images of human ileal tissue paraffin sections, stained for PHA-L (red), sWGA (green), Hoechst (blue), and merged image. Samples were incubated with fluorescein dyes (PHA-L 639, sWGA 488, and Hoechst 405), 10 μg/mL, in blocking buffer for 1 h at room temperature. Scale bar: 50 μm. n = 3–4. Fluorescence intensity of sWGA and L-PHA normalized to Hoechst was measured using Metamorph to compare healthy individuals and those with Crohn disease (CD). Ratio of sWGA/L-PHA also depicted. Individual data points, mean, and SEM graphed with statistical significance are determined by t test, with p value indicated by two asterisks (<0.01) or three asterisks (<0.001). (C) Volcano plot of glycosyltransferase genes differentially expressed in ileal mucosal biopsies from male and female pediatric patients with severe ileal Crohn disease compared to non-IBD controls. This is a secondary analysis of RNA-seq data (GEO: GSE57945 ). n = 42 controls, n = 63 CD patients. There is an enrichment for MGAT-related pathway members with increased expression of MGAT1 and decreased expression of MGAT3 , MGAT4A/4B , and MGAT5 . MGAT4B is the most highly expressed gene .

Article Snippet: After blocking, samples were incubated with fluorescein dyes ( Phaseolus vulgaris leucoagglutinin [PHA-L] rhodamine-conjugated lectin and succinylated wheat germ agglutinin [WGA] lectin) from Vector Labs (catalog nos.

Techniques: Glycoproteomics, Generated, Immunofluorescence, Microscopy, Staining, Incubation, Blocking Assay, Fluorescence, RNA Sequencing, Expressing

Journal: Human Genetics and Genomics Advances

Article Title: Aberrant N-glycosylation may be a therapeutic target in carriers of a common and highly pleiotropic variant in the manganese transporter ZIP8

doi: 10.1016/j.xhgg.2025.100517

Figure Lengend Snippet: Truncated sWGA + N-glycans are increased at the apical/brush border of ileal epithelial cells in ZIP8 391-Thr carriers with Crohn disease Representative lectin immunofluorescence images of ileal biopsies of genotyped patients with active Crohn’s ileitis in (A) ZIP8 391A/391A (non-carriers) and (B) ZIP8 391A/391T (ZIP8 391-Thr heterozygous carriers). Confocal laser-scanning triple-label immunofluorescence microscopy images of human ileal tissues paraffin sections, stained for L-PHA (red), sWGA (green), Hoechst (blue), and merged image. Samples were incubated with fluorescein dyes (L-PHA 639, sWGA 488, and Hoechst 405), 10 μg/mL, in blocking buffer for 1 h at room temperature. Fluorescence intensity of sWGA and L-PHA normalized to Hoechst was measured using Metamorph to compare healthy individuals and those with Crohn disease (CD). Scale bar: 50 μm. Asterisk highlights enhanced localization of sWGA staining to the apical membrane/glycocalyx of epithelial cells. To focus on the epithelial compartment, sWGA and L-PHA distribution and overlap were blindly scored by two investigators. Representative images from n = 3 patients/genotype included in figure; histology reviewed and scored for n = 9 ZIP8 391A/391A and n = 6 ZIP8 391A/391T individuals. ns, not statistically significant.

Article Snippet: After blocking, samples were incubated with fluorescein dyes ( Phaseolus vulgaris leucoagglutinin [PHA-L] rhodamine-conjugated lectin and succinylated wheat germ agglutinin [WGA] lectin) from Vector Labs (catalog nos.

Techniques: Immunofluorescence, Microscopy, Staining, Incubation, Blocking Assay, Fluorescence, Membrane

Journal: Human Genetics and Genomics Advances

Article Title: Aberrant N-glycosylation may be a therapeutic target in carriers of a common and highly pleiotropic variant in the manganese transporter ZIP8

doi: 10.1016/j.xhgg.2025.100517

Figure Lengend Snippet: ZIP8 391-Thr-associated defect in N-glycosylation in the ileal epithelial compartment is recapitulated in Zip8 393T-knockin mice (A) Confocal laser-scanning triple-label immunofluorescence microscopy images of Zip8 +/+ (WT), Zip8 +/393T (Het), and Zip8 393T/393T (HM) ileal tissues paraffin sections, stained for L-PHA (red), sWGA (green), Hoechst (blue), and merged image. Samples were incubated with fluorescein dyes (L-PHA 639, sWGA 488, and Hoechst 405), 10 μg/mL, in blocking buffer for 1 h at room temperature. Scale bar: 50 μm. N = 5 male and female mice/genotype with n = 3–5 fields of view/mice imaged. (B) Quantification of sWGA and L-PHA fluorescence intensity normalized to Hoechst measured using Metamorph. Statistical significance determined by one-way ANOVA, p value indicated by one asterisk (<0.05) or three asterisks (<0.001). (C) Averaged spectra of matrix-associated laser desorption/ionization (MALDI) mass spectrometry imaging (MSI) following on-tissue PNGase F digest to measure the differential abundance of N-glycan species in transverse section of distal ileal tissue of Zip8 +/+ and Zip8 393T/393T mice. Higher m/z species represent tri- and tetra-antennary N-glycan branching. N = 4 male mice/genotype. (D) Confocal laser-scanning triple-label immunofluorescence microscopy images of Jackson C57BL/6 male mice fed purified diet containing variable Mn (<1 ppm = Mn deficient and 2,400 ppm = Mn excess) for 4 weeks. Ileal tissues paraffin sections, stained for L-PHA (red), sWGA (green), Hoechst (blue), and merged image. N = 3 male mice in each group, 2–3 fields of view imaged and quantified per mouse. Scale bar: 50 μm. Individual data points, mean, and SEM graphed, with statistical significance determined by one-way ANOVA and p value indicated by two asterisks (<0.01) or three asterisks (<0.001).

Article Snippet: After blocking, samples were incubated with fluorescein dyes ( Phaseolus vulgaris leucoagglutinin [PHA-L] rhodamine-conjugated lectin and succinylated wheat germ agglutinin [WGA] lectin) from Vector Labs (catalog nos.

Techniques: Glycoproteomics, Knock-In, Immunofluorescence, Microscopy, Staining, Incubation, Blocking Assay, Fluorescence, Mass Spectrometry, Imaging, Purification

Journal: Human Genetics and Genomics Advances

Article Title: Aberrant N-glycosylation may be a therapeutic target in carriers of a common and highly pleiotropic variant in the manganese transporter ZIP8

doi: 10.1016/j.xhgg.2025.100517

Figure Lengend Snippet: Oral GlcNAc supplementation restores complex N-glycan branching in intestinal epithelial cells in Zip8 +/393T and Zip8 393T/393T mice Confocal laser-scanning triple-label immunofluorescence microscopy images of Zip8 +/+ (WT) (A), Zip8 +/393T (Het) (B), and Zip8 393T/393T (HM) (C) mice ileal tissue paraffin sections, stained for sWGA (green), L-PHA (red), Hoechst (blue), and merged image. Samples were incubated with fluorescein dyes (L-PHA 639, sWGA 488, and Hoechst 405), 10 μg/mL, in blocking buffer for 1 h at room temperature. L-PHA and sWGA fluorescence intensity measured using Metamorph. Scale bar: 50 μm. N = 4–5 male and female mice/genotype, with n = 3–7 fields of view/mice imaged. Individual data points, mean, and SEM are graphed. Statistical significance was determined by Kruskal-Wallis one-way ANOVA with Dunn’s multiple comparisons testing; p value indicated by one asterisk (<0.05), two asterisks (<0.01), three asterisks (<0.001), or four asterisks (<0.0001).

Article Snippet: After blocking, samples were incubated with fluorescein dyes ( Phaseolus vulgaris leucoagglutinin [PHA-L] rhodamine-conjugated lectin and succinylated wheat germ agglutinin [WGA] lectin) from Vector Labs (catalog nos.

Techniques: Glycoproteomics, Immunofluorescence, Microscopy, Staining, Incubation, Blocking Assay, Fluorescence