Journal: Cell Death & Disease

Article Title: Post-irradiation dietary restriction impairs hematopoiesis via inhibition of the pentose phosphate pathway in hematopoietic stem and progenitor cells

doi: 10.1038/s41419-025-08249-w

Figure Lengend Snippet: A – E Mice were subjected to 4.5 Gy X-ray irradiation and subsequently fed with AL diet or DR diet. BM cells or c-Kit + HSPCs were harvested at the indicated time points for q RT-PCR analysis. NIR mice receiving an AL diet were monitored as a control. Relative expression of indicated genes involving DNA damage and repair were analyzed, with β-actin as the internal control ( n = 5 mice per group from 1 experiment representative of 2 independent experiments). F , G Apoptosis rates in HSCs at indicated time points post-irradiation and in NIR mice, as determined by FACS ( F ). Representative FACS plots at day 1 post-irradiation ( G ) ( n = 5 mice per group from 1 experiment, representative of 2 independent experiments). H , I Quantification and representative images of γH2AX and 53BP1 colocalization in c-Kit + HSPCs at indicated time points post-irradiation and before irradiation, determined by immunofluorescent staining (scale bar: 10 μm) ( n = 5 mice per group from 1 experiment representative of 2 independent experiments). J – L Quantification and representative images of comet assay tail DNA in c-Kit + HSPCs at indicated time points post-irradiation and before irradiation (scale bar: 10 μm) ( n = 5 mice per group from 1 experiment, representative of 2 independent experiments). M Relative expression of G6PD mRNA in BM cells of AL and DR mice at the indicated time points post-irradiation and NIR AL mice, analyzed by qRT-PCR with β-actin as the internal control ( n = 5 mice per group from 1 experiment representative of 2 independent experiments). Dynamic measurements by ELISA of G6PD enzyme activity ( N ) and NADPH ( O ) content in BM cells of AL and DR mice at the indicated time points under IR or no-conditions conditions. P G6PD enzyme activity and Q NADPH content in BM cells of AL and DR mice given 5% sucrose water or normal drinking water at NIR/IR 14 days ( n = 5 mice per group from 1 experiment representative of 2 independent experiments). R Average fluorescence intensity of ROS in BM cells of AL and DR mice at indicated time points under non-irradiated or irradiated conditions ( n = 5 mice per group from 1 experiment representative of 2 independent experiments). Relative expression of indicated genes involving ROS in BM cells ( S ) and c-Kit + HSPCs ( T ) of AL and DR mice at indicated time points post-irradiation, analyzed by qRT-PCR with NIR AL mice as a control ( n = 5 mice per group from 1 experiment representative of 2 independent experiments). U and V Quantification and representative images of Nrf2 foci in the nucleus of BM cells of AL and DR mice at 5 h post-irradiation and under non-irradiated conditions, determined by immunofluorescent staining (scale bar: 10 μm) ( n = 5 mice per group from 1 experiment representative of 2 independent experiments). Results were displayed as mean ± SD;. ns, not significant; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001 by one-way ANOVA test and Two-way ANOVA test with Tukey’s multiple comparisons test. HSPCs: hematopoietic stem and progenitor cells. The purple symbols indicate statistical significance between the non-irradiated groups. The green symbols indicate statistical significance between the irradiated groups.

Article Snippet: The G6PD activator AG1 was purchased from MedChemExpress Co., Ltd., China (CAS No. 421581-52-4; Lot number: HY-123962; purity: 99.54%), and was diluted in dimethyl sulfoxide (DMSO) for injection intraperitoneally every other day for 9 times post-irradiation.

Techniques: Irradiation, Reverse Transcription Polymerase Chain Reaction, Control, Expressing, Staining, Single Cell Gel Electrophoresis, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Activity Assay, Fluorescence

Journal: Cell Death & Disease

Article Title: Post-irradiation dietary restriction impairs hematopoiesis via inhibition of the pentose phosphate pathway in hematopoietic stem and progenitor cells

doi: 10.1038/s41419-025-08249-w

Figure Lengend Snippet: A Experimental scheme. Wild-type C57BL/6 mice (2 months old) were irradiated with 4.5 Gy X-rays and fed with an AL diet. Mice were then injected intraperitoneally with 10 mg/kg of 6-aminonicotinamide (6-AN) or saline daily for 7 days post-irradiation, and were continued on an AL diet until 1 month post-irradiation ( n = 5 mice per group from 1 experiment representative of 2 independent experiments). G6PD enzyme activity ( B ) and NADPH content ( C ) in BM cells of AL mice 1 month post-irradiation, analyzed using ELISA. WBC ( D ), Neutrophil ( E ), and lymphocyte ( F ) counts in PB of AL mice 1 month post-irradiation. G , H Spleen and thymus weights of AL mice 1 month post-irradiation. I Total BM cell counts of AL mice 1 month post-irradiation. FACS analysis of frequencies of B220 + lymphocytes in PB ( J ), spleen ( K ), BM ( L ); and frequencies of HSCs ( M ), CMPs ( O ), and GMPs ( P ), and representative FACS Plots of HSCs ( N ) and CMPs/GMPs/MEPs ( Q ) of AL mice 1 month post-irradiation. R and S Quantification and representative images of γH2AX and 53BP1 colocalization in BM cells of AL mice 1 month post-irradiation, determined by immunofluorescent staining (scale bar: 10 μm). T–A’ Chimerism of donor-derived cells in the PB, including WBCs ( T ), B cells ( V ), T cells ( X ), and myeloid cells ( Z ) chimerism after transplantation at indicated time points and representative FACS plot of WBCs ( U ), B cells ( W ), T cells ( Y ), myeloid cells ( A’ ) at 4 months post-transplantation. Results were displayed as mean ± SD;. ns, not significant; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001 by unpaired two-tailed Student’s t -test and Two-way ANOVA test with Tukey’s multiple comparisons test. IR AL + 6-AN: irradiated ad libitum group injected with 6-AN; IR AL + NS: irradiated ad libitum group injected with saline.

Article Snippet: The G6PD activator AG1 was purchased from MedChemExpress Co., Ltd., China (CAS No. 421581-52-4; Lot number: HY-123962; purity: 99.54%), and was diluted in dimethyl sulfoxide (DMSO) for injection intraperitoneally every other day for 9 times post-irradiation.

Techniques: Irradiation, Injection, Saline, Activity Assay, Enzyme-linked Immunosorbent Assay, Staining, Derivative Assay, Transplantation Assay, Two Tailed Test

Journal: Cell Death & Disease

Article Title: Post-irradiation dietary restriction impairs hematopoiesis via inhibition of the pentose phosphate pathway in hematopoietic stem and progenitor cells

doi: 10.1038/s41419-025-08249-w

Figure Lengend Snippet: A Experimental scheme. Wild-type C57BL/6 mice (2 months old) were irradiated with 4.5 Gy X-rays, fed with a DR diet. Mice were then injected intraperitoneally with 20 mg/kg of AG1 or DMSO every other day for 9 times post-irradiation, and were continued on DR until 1 month post-irradiation ( n = 5 mice per group from 1 experiment representative of 2 independent experiments). G6PD enzyme activity ( B ) and NADPH content ( C ) in BM cells of DR mice 1 month post-irradiation, analyzed using ELISA. WBC ( D ), Neutrophil ( E ), and lymphocyte ( F ) counts in PB of DR mice 1 month post-irradiation. G , H Spleen and thymus weights of DR mice 1 month post-irradiation. I Total BM cell counts of DR mice 1 month post-irradiation. FACS analysis of frequencies of B220 + lymphocytes in PB ( J ), spleen ( K ), BM ( L ); and frequencies of HSCs ( M ), CMPs ( O ) and GMPs ( P ), and representative FACS Plots of HSCs ( N ) and CMPs/GMPs/MEPs ( Q ) of DR mice 1 month post-irradiation. R and S Quantification and representative images of γH2AX and 53BP1 colocalization in BM cells of DR mice 1 month post-irradiation, determined by immunofluorescent staining (scale bar: 10 μm). Chimerism of donor-derived cells in the PB, including WBCs ( T ), B cells ( V ), T cells ( X ), and myeloid cells ( Z ) chimerism after transplantation at indicated time points and representative FACS plot of WBCs ( U ), B cells ( W ), T cells ( Y ), myeloid cells ( A’ ) at 4 month post-transplantation. Results were displayed as mean ± SD;. ns, not significant; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001 by unpaired two-tailed Student’s t test, and Two-way ANOVA test with Tukey’s multiple comparisons test. IR DR + AG1: irradiated dietary restriction group injected with AG1; IR DR + DMSO: irradiated dietary restriction group injected with DMSO.

Article Snippet: The G6PD activator AG1 was purchased from MedChemExpress Co., Ltd., China (CAS No. 421581-52-4; Lot number: HY-123962; purity: 99.54%), and was diluted in dimethyl sulfoxide (DMSO) for injection intraperitoneally every other day for 9 times post-irradiation.

Techniques: Irradiation, Injection, Activity Assay, Enzyme-linked Immunosorbent Assay, Staining, Derivative Assay, Transplantation Assay, Two Tailed Test

Journal: Cell Death & Disease

Article Title: Post-irradiation dietary restriction impairs hematopoiesis via inhibition of the pentose phosphate pathway in hematopoietic stem and progenitor cells

doi: 10.1038/s41419-025-08249-w

Figure Lengend Snippet: A Experimental scheme. Wild-type C57BL/6 mice (2 months old) were irradiated with 4.5 Gy X-rays, fed with either AL diet or DR diet or DR diet 1 week, then AL feeding 3 weeks (daily food intake restricted to 70% of the intake of age- and sex-matched AL mice). NIR mice receiving an AL or DR diet were also monitored as a control ( n = 5 mice per group from 1 experiment representative of 2 independent experiments. B and C G6PD enzyme activity ( B ) and NADPH content ( C ) in BM cells of RF mice 1 month post-irradiation, analyzed using ELISA. WBC ( D ), Neutrophil ( E ), and lymphocyte ( F ), RBC ( G ), platelet ( H ) counts in PB of RF mice 1 month post-irradiation. I , J Spleen and thymus weights of RF mice 1 month post-irradiation. K Total BM cell counts of RF mice 1 month post-irradiation. FACS analysis of frequencies of B220 + lymphocytes in PB ( L ), spleen ( M ), BM ( N ); and frequencies of HSCs ( O ), CMPs ( P ) and GMPs ( Q ), MEPs ( R ), and RMBMs ( S ) of RF mice 1 month post-irradiation. T Apoptosis rates in HSCs of RF mice 1 month post-irradiation. U Average fluorescence intensity of ROS in BM cells of RF mice 1 month post-irradiation. V and W Quantification and representative images of γ-H2AX and 53BP1 colocalization in BM cells of RF mice 1 month post-irradiation, determined by immunofluorescent staining (scale bar: 10 μm). Results were displayed as mean ± SD; ns, not significant; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001 by one-way ANOVA test with Tukey’s multiple comparisons test. IR RF: irradiated refeeding group.

Article Snippet: The G6PD activator AG1 was purchased from MedChemExpress Co., Ltd., China (CAS No. 421581-52-4; Lot number: HY-123962; purity: 99.54%), and was diluted in dimethyl sulfoxide (DMSO) for injection intraperitoneally every other day for 9 times post-irradiation.

Techniques: Irradiation, Control, Activity Assay, Enzyme-linked Immunosorbent Assay, Fluorescence, Staining

Journal: BMC Infectious Diseases

Article Title: High daily dose Short COurse PrimaquinE after G6PD testing for the radical cure of Plasmodium vivax malaria in Indonesia and Papua New Guinea: the SCOPE implementation study protocol

doi: 10.1186/s12879-025-11109-9

Figure Lengend Snippet: Schedule of enrolment, interventions, and assessments 1 Malaria diagnosis is part of routine clinic procedures and takes place prior to study enrolment 2 Day 3 review may be a phone call if the CHW cannot access the patient at home, or at the clinic if the patient presents there 3 Unplanned visits are defined as any visit to the health facility or hospital (outside of scheduled day 3 or day 7 visits) during or within 10 days of completing treatment 4 During Stage 2 200 consecutive febrile patients at each clinic will be consented for the post-implementation survey, which includes the following procedures: Hb, G6PD and Microscopy only. If positive for vivax malaria, patients will also be recruited to Stage 2 5 Microscopy will only be performed for 200 consecutive febrile patients at each clinic during post-implementation survey, not Stage 2 6 Any AESI/SAE identified during study participants’ primaquine treatment will be captured

Article Snippet: The SD Biosensor provides reliable differentiation of patients with deficient (< 30%), intermediate (30- < 70%) and normal (≥ 70%) G6PD activity within 2 min using one drop of capillary blood, therefore lending itself to pre-treatment testing.

Techniques: Biomarker Discovery, Microscopy

Journal: bioRxiv

Article Title: Metabolic control of enteroendocrine cell fate through a redox state sensor CtBP

doi: 10.1101/2025.06.30.662346

Figure Lengend Snippet: (A) Schematic presentation of the BiFC system to analyze CtBP homodimerization by the activity of metabolic pathways or enzymes influencing the redox balance of nicotinamide cofactors. GAL4 expression (Pros-GAL4 ts ) was activated 16 h prior measurement for all BiFC experiments. (B) CtBP dimerization as measured by BiFC reporters expressed in EE cells (Pros-GAL4 ts >CtBP-CC OE :CtBP-VN OE ), EE cells marked by anti-Pros antibodies. (C) CtBP dimerization (number of BiFC positive cells within EE cell population) is inhibited by LbNOX and TpNOX overexpression, which oxidize NADH and NADPH, respectively. The data are analyzed using ordinary one-way ANOVA, ****P < 0.0001. ns indicates no significant difference in the studied parameter. (D) CtBP homodimerization in EE cells (BiFC) is regulated by sugar catabolism and nicotinamide metabolism. A diet containing 2-deoxyglucose (DG) was used to inhibit glucose catabolism. Diets with NR and AG1, a small molecule activator of the enzyme glucose-6-phosphate dehydrogenase (G6PD), were used to modulate the metabolic state. The data are analyzed using one-way ANOVA with Kruskal-Wallis correction for not normal distribution, with *P=0.0143, ***P = 0.0002, ****P < 0.0001. ns indicates no significant difference in the studied parameter. (E) Glucose catabolism through glycolysis and the pentose phosphate pathway activates CtBP homodimerization in the EE cells. The enzymes of glycolysis (phosphofructokinase, pfk ) and the pentose phosphate pathway (G6PD, Zw ) were depleted in EE cells. The data are analyzed using ordinary one-way ANOVA, with *P=0.0264, ****P < 0.0001. (F) Depletion of pfk and Zw in larval EE cells leads to sugar intolerance. Larval development on HSD (30% sucrose) was fully impaired with pfk knockdown (Pros-GAL4>) (χ = 128.0, P < 0.0001) and modestly delayed by Zw knockdown (χ = 12.1, P = 0.0004), as determined by the log-rank test. (G) Knockdown of pfk and Zw (Pros-GAL4>) in larvae fed on moderate (MSD, 5% sucrose) or high sugar diet (HSD, 30% sucrose) reduces the proportion of EE cells in the larval intestine. These genetic manipulations also decrease populations of Dh31-positive (H) and Tk-positive (I) cells (unpaired t-test, with **P < 0.01, ***P < 0.001).

Article Snippet: NR and AG1 diet contained yeast in concentration of 15% (w/v) and 500 μM nicotinamide riboside chloride (Sigma-Aldrich, SMB00907) or 10 μM glucose-6-phosphate dehydrogenase activator AG1 (MedChemExpress, HY-123962).

Techniques: Activity Assay, Expressing, Over Expression, Knockdown