Journal: Current Opinion in Gastroenterology

Article Title: Transoral incisionless fundoplication: current status

doi: 10.1097/MOG.0000000000000275

Figure Lengend Snippet: Mean quality of life scores before transoral incisionless fundoplication and at 6 and 12-month follow-up in patient's randomized to the transoral incisionless fundoplication group (TEMPO trial) [ 4 ▪ , 5 ▪ ]

Article Snippet: This may be explained by the unfair comparison of TIF to the first generation of GERD endoluminal therapies (e.g., EndoCinch, Enteryx, Gatekeeper, and Endoscopic Plicator System, NDO Surgical, INC, Mansfield, Massachusetts, USA), which, with the exception of the Stretta procedure, are no longer available because of safety concerns or lack of effectiveness.

Techniques:

Journal: Redox Biology

Article Title: Redox signaling-mediated S-glutathionylation of protein disulfide isomerase A1 initiates intrinsic apoptosis and contributes to accelerated aging

doi: 10.1016/j.redox.2025.103680

Figure Lengend Snippet: CSE induces S-glutathionylation of PDIA1. (A, B) Recombinant PDI proteins were treated by 20 % CSE and 5 mM GSH at 37 °C for 30 min, part of each sample was separated on a non-reducing gel and probed for the presence of S-glutathionylation (A), the other part was desalted by Biospin 6 columns. The protein was digested with endoproteinase LysC, the digest desalted, and the peptides subjected to LC-MS/MS identification (B). Spectra of domain a (left) and a’ (right) are shown. C fragment ions (from the peptide N-terminal) obtained by electron transfer dissociation (ETD) are shown in blue; while Z fragments (from the peptide C-terminal) are shown in red. The glutathione diagnostic fragment is shown in green. The glutathionylated cysteines are shown with an ∗ (C) A docked model of PDIA1 WT and PDIA1 FLFL with GSH, based on the human PDIA1 domain a’ (PDB ID: 3UEM ), showing disrupted interactions in the mutant version. His399 and Lys401 of PDIA1 WT interact with GSH, but these interactions are lost in PDIA1 FLFL , where His399 and Lys401 are replaced by Phe and Leu, respectively. (D) Fluorescent FRET-based analysis showing PDIA1 and GSTP1 interaction. The Y-axis represents the decrease in Alexa546 emission, indicating GSTP1 binding to PDIA1 WT or PDIA1 FLFL . The X-axis represents the total amount of GSTP1 labeled with QSY35. Data represent three independent experiments. (E) PDIA1 was immunoprecipitated from MLE12 cells, and GSTP1 was detected as co-immunoprecipitating with PDIA1 in three independent experiments. ( F ) Docked models of PDIA1 WT and PDIA1 FLFL with GSTP1, based on the human PDIA1 (PDB ID: 3UEM ) and GSTP1 (PDB ID: 6GSS ). GSH in this model is adapted from the 6GSS structure. PDIA1 WT Lys401 interacts with GSTP1 Gln51, and His399 is close to GSTP1 Asp94. These interactions are disrupted in PDIA1 FLFL , where His399 and Lys401 are replaced by Phe and Leu. ( G-H ) The model structure of wild type versus H399F, K401L PDIA1 and their residue-specific interaction map with GSTP1 build using Ligplot software. Please note that changes in the interaction map of His399Phe and Lys401Leu (PDIA1 WT → PDIA1 FLFL ) weakens interaction with GSTP1 and in GSH repositioning. The direct ionic interactions between PDIA1-His399 and GSTP1-Asp94, and PDIA1-Lys401 and GSTP1-Gln51 do not occur in PDIA1 FLFL . Moreover, the carboxyl oxygen of PDIA1-Trp396 interacts with GSTP1-Gln64 in PDIA1 WT and is replaced by an interaction with the side chain of GSTP1-Arg13 in the case of PDIA1 FLFL , compromising interactive interphase between those protein. Direct interaction of PDIA1-Lys401 with oxygen of GSH is abolished as well.

Article Snippet: A high-capacity cDNA reverse transcription kit (Applied Biosystems) was used to generate cDNA. qPCR was performed using the TaqMan universal master mix II, no UNG (Life Technologies), with the following assays: Mm00496606_m1 (GSTP1) and Mm99999915_g1 (GAPDH).

Techniques: Recombinant, Liquid Chromatography with Mass Spectroscopy, Diagnostic Assay, Mutagenesis, Binding Assay, Labeling, Immunoprecipitation, Residue, Software

Journal: Redox Biology

Article Title: Redox signaling-mediated S-glutathionylation of protein disulfide isomerase A1 initiates intrinsic apoptosis and contributes to accelerated aging

doi: 10.1016/j.redox.2025.103680

Figure Lengend Snippet: Inhibiting GSTP1-mediated S-glutathionylation prevents PDIA1 relocation to ERMCS upon CSE exposure. MLE12 cells were treated as indicated: (i) treated with GSTP1 inhibitor TLK199, or (ii) transfected with siRNA-Control/siRNA-GSTP1 for 24 h. Prior to experiments, cells were loaded with Mito-tracker-red and exposed, or not, to 15 min of 20 % CSE. PDIA1 (Abberior STAR-488), mitochondria (Mito-tracker-red), and nuclei (Hoechst) were visualized. (A, H) Representative single-plane images. (B, I) Enlarged images from boxed areas in (A). Arrows indicate PDIA1-mitochondria co-localization in CSE-treated cells. (C, J) Further enlarged images from boxed areas in (B) with line scans crossing mitochondria and PDIA1. (D, K) Graph showing line scan data with relative fluorescence intensity per pixel. Arrows indicate co-localization regions. (E, L) Pearson's coefficient calculated from boxed areas in (A) using the Coloc2 function in FUJI ImageJ software. (F, M) Graph summarizing Pearson's coefficient from three independent experiments. Significant differences ( p < 0.001) between control and CSE-exposed cells with inhibited S-glutathionylation indicate partial but not complete rescue. Statistical comparisons are detailed in Sup. .

Article Snippet: A high-capacity cDNA reverse transcription kit (Applied Biosystems) was used to generate cDNA. qPCR was performed using the TaqMan universal master mix II, no UNG (Life Technologies), with the following assays: Mm00496606_m1 (GSTP1) and Mm99999915_g1 (GAPDH).

Techniques: Transfection, Control, Fluorescence, Software

Journal: Redox Biology

Article Title: Redox signaling-mediated S-glutathionylation of protein disulfide isomerase A1 initiates intrinsic apoptosis and contributes to accelerated aging

doi: 10.1016/j.redox.2025.103680

Figure Lengend Snippet: IMC profiling of lung parenchyma reveals the impact of aging and cigarette smoke exposure on PDIA1, GSTP1, and mitochondrial bioenergetic markers in AEC2s. (A) Representative IMC images of 14-week-old non-smoker (NS) mouse lung parenchyma using a 10-marker panel. Shown are proSPC (164Dy; AEC2 marker), PDIA1 (170Er), GSTP1 (168Er), and nuclear DNA (191Ir and 193Ir). Yellow arrows indicate representative AEC2. Yellow arrows point to the representative AEC2. ( B, D-F ) Single-cell quantitative analysis of marker expression across conditions. ( B ) PDIA1 relative intensity in AEC1s and AEC2. Each dot represents a single cell. ( C) Schematic outlining the experimental timeline and cigarette smoke (CS) exposure protocol. C57Bl/6 mice of different ages were exposed or not exposed to CS for defined durations. ( D–E) Violin plots showing single-cell quantification of relative signal intensity per AEC2 for PDIA1 (D) and GSTP1 (E) across NS and smoker (SM) groups. ( F ) Quantification of AEC2 abundance, expressed as the percentage of AEC2s per region of interest (ROI; 500 × 500 μm), across experimental groups. Data are presented as mean ± standard deviation from three biological replicates (N = 3 mice per group), each with technical triplicates (3 Regions of Interests (ROIs) per mouse). Final statistical comparisons were performed using unpaired two-tailed t-tests. P-values <0.05 were considered statistically significant.

Article Snippet: A high-capacity cDNA reverse transcription kit (Applied Biosystems) was used to generate cDNA. qPCR was performed using the TaqMan universal master mix II, no UNG (Life Technologies), with the following assays: Mm00496606_m1 (GSTP1) and Mm99999915_g1 (GAPDH).

Techniques: Marker, Expressing, Standard Deviation, Two Tailed Test

Journal: Redox Biology

Article Title: Redox signaling-mediated S-glutathionylation of protein disulfide isomerase A1 initiates intrinsic apoptosis and contributes to accelerated aging

doi: 10.1016/j.redox.2025.103680

Figure Lengend Snippet: S-glutathionylated PDIA1 causes Cyt c release, apoptosis, and cytotoxicity following CSE exposure . (A – C) Mitochondria were incubated with CSE (20 %)-treated PDIA1 WT or PDIA1 FLFL , with or without CsA and Ca 2+ (150 μM), in the presence of cytosol for 30 min at 37 °C. (A) Mitochondria were pelleted, and the presence of Cyt c in the supernatant was evaluated by Western blotting. PDH antibodies were used to assess mitochondrial input as a loading control. (B) Quantification of Cyt c release relative to control (mean ± SD, ∗∗∗ p < 0.001). (C) Cyt c release after incubation with S-glutathionylated PDIA1 induced caspase activation, measured via DEVD-AFC cleavage after 20 min. (D – F) Caspases are not required for S-glutathionylated PDIA1-induced Cyt c release. Mitochondria were incubated with CSE (20 %)-treated PDIA1 WT or PDIA1 FLFL , with or without zVAD-fmk (10 μM) and Ca 2+ (150 μM), in the presence of cytosol for 30 min at 37 °C. (D) Cyt c levels in the cytosol were analyzed by Western blot. PDH antibodies were used to assess mitochondrial input as a loading control. (E) Relative Cyt c release (mean ± SD, ∗∗∗ p < 0.001). (F) Caspase activity measured via DEVD-AFC cleavage (∗∗∗ p < 0.001). (G) S-glutathionylated PDIA1 enhances CSE-induced cytotoxicity, assessed by MTT assay in MLE12 cells transfected with EV, PDIA1 WT , or PDIA1 FLFL . (H) MTT assay measuring CSE cytotoxicity with or without the GSTP1 inhibitor TLK199.

Article Snippet: A high-capacity cDNA reverse transcription kit (Applied Biosystems) was used to generate cDNA. qPCR was performed using the TaqMan universal master mix II, no UNG (Life Technologies), with the following assays: Mm00496606_m1 (GSTP1) and Mm99999915_g1 (GAPDH).

Techniques: Incubation, Western Blot, Control, Activation Assay, Activity Assay, MTT Assay, Transfection

Journal: Redox Biology

Article Title: Redox signaling-mediated S-glutathionylation of protein disulfide isomerase A1 initiates intrinsic apoptosis and contributes to accelerated aging

doi: 10.1016/j.redox.2025.103680

Figure Lengend Snippet: A schematic illustration of the present report. We identify GSTP1 as a stress-dependent regulator of ER–mitochondria contact site (ERMCS) composition. GSTP1 modulates the S-glutathionylation of PDIA1, which in turn dictates the intra-ER localization of distinct PDIA1 pools, ultimately influencing intraorganellar communication.

Article Snippet: A high-capacity cDNA reverse transcription kit (Applied Biosystems) was used to generate cDNA. qPCR was performed using the TaqMan universal master mix II, no UNG (Life Technologies), with the following assays: Mm00496606_m1 (GSTP1) and Mm99999915_g1 (GAPDH).

Techniques: