Blog

  • Japan 0-0 Wales: Watch, listen & follow live

    Japan 0-0 Wales: Watch, listen & follow live

    Japan team news: Leitch to lead inexperienced sidepublished at 05:42 British Summer Time

    Japan v Wales (06:00 BST)

    Image source, Getty Images
    Image caption,

    Michael Leitch has featured in four World Cups and played 87 internationals

    Eddie Jones has named eight uncapped internationals in his matchday squad, with around 15 players unavailable.

    Michael Leitch, 36, will captain the side and is the only member of the squad who has more than 50 caps.

    Prop Yota Kamimori and wing Kippei Ishida, who Jones says can be a Japan version of South Africa double World Cup winner Cheslin Kolbe, will both win first caps, while there are six new potential caps on the bench.

    Toulouse scrum-half Naoto Saito, who was part of the squad that won the French Top 14 title last weekend, is not involved today, but will be available for the second Test in Kobe.

    Japan: Takuro Matsunaga; Kippei Ishida, Dylan Riley, Shogo Nakano, Malo Tuitama; Seungsin Lee, Shinobu Fujiwara; Yota Kamimori, Mamoru Harada, Shuhei Takeuchi, Epineri Uluiviti, Warner Deans, Michael Leitch (capt), Jack Cornelsen, Amato Fakatava.

    Replacements: Hayate Era, Sena Kimura, Keijiro Tamefusa, Waisake Raratubua, Ben Gunter, Shuntaro Kitamura, Ichigo Nakakusu, Halatoa Vailea.

    Continue Reading

  • Identification of Shared Biomarkers in Chronic Kidney Disease and Diab

    Identification of Shared Biomarkers in Chronic Kidney Disease and Diab

    Introduction

    The global prevalence of chronic kidney disease (CKD) has been rising steadily, currently affecting approximately 10.8% of the total population.1 CKD is characterized by abnormalities in kidney structure or function persisting for over 3 months, affecting overall health. A key indicator is a glomerular filtration rate (GFR) of less than 60 mL/ (min·1.73 m²), accompanied by at least one of the following markers of kidney injury: albuminuria, abnormal urinary sediment (eg, hematuria), electrolyte disturbances due to renal tubule dysfunction, histological abnormalities, structural changes in imaging, or a history of kidney transplantation.2 CKD often presents gradually, with subtle or atypical symptoms in the early stages, making timely detection challenging. At onset, typical indicators include hypertension, hyperglycemia, and microalbuminuria, which are not highly sensitive to standard diagnostic tests, contributing to a poor clinical prognosis.

    As the condition advances, it can evolve into nephrotic syndrome, chronic nephritis, or acute nephritis, with some patients progressing to end-stage renal disease (ESRD). The long waiting period for kidney transplantation, due to limited donor availability, results in many patients with ESRD relying on dialysis for survival. Over 60% of these patients undergo dialysis for more than a year, with approximately 23% needing long-term dialysis for over 3 years.3 This not only imposes a significant physiological, psychological, and financial burden but also affects the quality of life.

    Treatment for CKD primarily focuses on slowing nephron damage, managing hyperfiltration, addressing complications, and providing renal replacement therapy. However, hemodialysis often leads to poor functional outcomes, including uremia-related malnutrition and muscle wasting, and carries risks of infection and vascular complications, further compromising patient quality of life. While kidney transplantation offers improved quality of life, recipients face persistent CKD-related symptoms and complications from immunosuppressive therapies.4 Consequently, more effective and scientifically-based methods are needed for the diagnosis and treatment of CKD.

    Diabetic nephropathy (DN) is a common and serious microvascular complication of diabetes, particularly prevalent in type 2 diabetes mellitus (T2DM), primarily induced by hyperglycemia. Its clinical manifestations include proteinuria, progressive renal dysfunction, hypertension, and edema. In China, approximately 20% to 40% of patients with diabetes are affected by DN, with most cases in the early, asymptomatic stages.5 Current research suggests that the pathogenesis of DN is closely associated with hyperglycemia, the accumulation of advanced glycosylation end products, as well as inflammatory and immune responses.6 Currently, DN has surpassed glomerulonephritis as the leading cause of new cases of CKD in China.7 As DN advances, patients face an increased risk of developing CKD due to factors such as RAAS activation and microvascular damage induced by sustained hyperglycemia and hypertension.8 Early screening and prompt diagnosis of DN are essential to prevent progression to CKD and end-stage nephropathy. However, the precise mechanisms by which DN contributes to CKD remain unclear. Thus, studying DN is vital not only for understanding the underlying mechanisms of CKD but also for identifying new therapeutic targets for both DN and CKD prevention and treatment.

    Single-cell RNA sequencing (scRNA-seq) is a cutting-edge high-throughput sequencing technique that enables the analysis of gene expression profiles at the single-cell level. By analyzing cellular composition, gene enrichment pathways, and intercellular communication, scRNA-seq offers insights into the underlying pathological processes of diseases. Recently, scRNA-seq has gained widespread use due to its sensitivity, accuracy, and efficiency. Unlike traditional sequencing methods, scRNA-seq enables detailed analysis of the cellular spectrum, identification of specific cell types, and mapping of gene expression patterns in heterogeneous cell samples. This allows for the study of gene expression at the single-cell level, providing a microscopic view of disease progression.9 The application of scRNA-seq in kidney research is promising, as it enhances understanding of cellular heterogeneity in CKD and DN, as well as identifying the potential mechanisms between these conditions. Additionally, scRNA-seq can help elucidate the correlation between CKD and DN, offering valuable insights for identifying biomarkers that can predict disease progression and inform patient-specific treatment strategies.10

    In this study, biomarkers associated with DN and CKD were identified through single-cell analysis, differential expression analysis, and protein-protein interaction (PPI) network construction. A comprehensive bioinformatics approach was utilized, including cell communication analysis, pseudotime series analysis, and gene set enrichment analysis (GSEA). The mechanisms of these biomarkers were examined, with an emphasis on key cell types and immune responses in patients with DN and CKD. This research provides a theoretical foundation and novel perspectives for studying disease associations, advancing diagnostic methods, and identifying therapeutic targets. Additionally, cellular-level validation of biomarker expression offers valuable insights for distinguishing between these two conditions.

    Materials and Methods

    Single-Cell RNA-Sequencing

    Nine blood samples were collected for scRNA-seq, including three control samples, three CKD samples (All were stage 5 chronic kidney disease), and three DN samples (One sample was stage 4 chronic kidney disease, and the rest were stage 5 chronic kidney disease, and all were stage 5 diabetic nephropathy). Sample grouping information is shown in Supplementary Table 1.

    Patients with DN were selected based on the 2020 Kidney Disease: Improving Global Outcomes (KDIGO) Guidelines, using the following criteria: (1) A urine albumin-creatinine ratio (UACR) of ≥ 30 mg/g, measured at least twice over a 3 to 6-month period, with other factors excluded; (2) An estimated glomerular filtration rate (eGFR) of < 60mL • min-1 • (1.73 m2) −1 persisting for more than 3 months; (3) Renal biopsy results indicating pathological changes consistent with DN. Patients with CKD met the 2020 KDIGO Guidelines for CKD diagnosis in the absence of diabetes. The exclusion criteria were: (1) Severe infections; (2) Malignant tumors; (3) Active autoimmune diseases; (4) Concurrent cardiovascular or cerebrovascular events; (5) Pregnancy. This study was approved by our hospital’s Ethics Committee, and informed consent was obtained from all participants.

    Chronic Kidney Disease is classified into stages based on GFR and albuminuria, as outlined by the KDIGO guidelines. The KDIGO 2012 Classification includes: Stage 1: GFR ≥ 90 mL/min/1.73 m² with evidence of kidney damage (eg, albuminuria, structural abnormalities, or genetic disorders); Stage 2: GFR 60–89 mL/min/1.73 m² with kidney damage; Stage 3a: GFR 45–59 mL/min/1.73 m²; Stage 3b: GFR 30–44 mL/min/1.73 m²; Stage 4: GFR 15–29 mL/min/1.73 m²; Stage 5: GFR < 15 mL/min/1.73 m² or kidney failure requiring dialysis/transplantation.11

    DN progression is classified based on GFR and albuminuria, integrating criteria from both diabetes and CKD guidelines. The KDIGO framework is widely used, with modifications specific to Diabetic Kidney Disease. Stage 1: GFR > 90 mL/min/1.73 m² (elevated due to renal hyperfiltration), early glomerular hypertrophy and hyperfiltration; Stage 2: GFR normal or mildly elevated (≥ 90 mL/min/1.73 m²), persistently elevated albumin-to-creatinine ratio (ACR 30–300 mg/g, moderately increased), kidney structural damage (eg, glomerular basement membrane thickening); Stage 3: GFR 60–89 mL/min/1.73 m² (CKD Stage 2), ACR ≥ 300 mg/g (severely increased), with clinical signs of hypertension and progressive proteinuria; Stage 4: GFR 15–59 mL/min/1.73 m² (CKD Stages 3–4), persistent ACR ≥ 300 mg/g, complications include declining kidney function, edema, and cardiovascular risks; Stage 5: kidney failure, GFR < 15 mL/min/1.73 m² (CKD Stage 5), requiring dialysis or transplantation.12

    Peripheral blood mononuclear cells (PBMCs) were isolated from peripheral blood samples using Ficoll density gradient centrifugation. Cell viability, assessed using AO/PI double fluorescent staining on a Countstar Rigel (S2) instrument, was required to exceed 85%. Following quality inspection, the single-cell suspension met the quality control criteria and proceeded with library construction, adhering to the SOP “ChromiumNextGEMSingleCell3_3.1_rev_d” from 10x Genomics. The Illumina Nova-seq 6000 PE150 platform was employed for sequencing the single-cell library.

    Data Filtering

    Sequencing data were initially examined for data volume, sequencing base quality, and sequencing saturation, followed by sequence statistics analysis using CellRanger (v 7.0).13 Single-cell analysis was then conducted on the RNA-sequencing dataset using the Seurat package (v 3.1.5).14 A Seurat object was created with parameters min.cells = 100 and min.features = 100 to filter out low-quality cells. Next, the scDblFinder package (v 1.17.2) was applied to identify and eliminate doublet cells.15 Cell screening criteria were as follows: library size exceeding 500 but below the 95th percentile (10,000 cells), gene counts below the 95th percentile (10,000 cells), and mitochondrial content restricted to less than 10%. Gene expression in each cell was normalized using the LogNormalize method.

    Principal Component Analysis (PCA) and Cell Annotation

    The FindVariableFeatures function with the variance-stabilizing transformation (vst) method was employed to identify genes exhibiting significant variation across cells. From this analysis, the top 2000 genes with the highest variability were selected. To minimize the effects of differing sequencing batches, data from the nine samples were integrated. The FindIntegrationAnchors function was used to identify anchors from a set of Seurat objects, and the IntegrateData function was then applied to merge the samples based on these anchors. PCA was applied to scale and reduce the data dimensionality. Principal components (PCs) with higher rankings in PCA encapsulate more diverse and valuable differential features. An elbow plot was constructed to identify the appropriate number of PCs for clustering analysis. Cells were clustered in an unsupervised manner using the FindNeighbors and FindClusters functions (resolution = 1). t-distributed stochastic neighbor embedding (t-SNE) was used to visualize cell clusters. Marker genes for each cluster were identified and annotated by comparing them with known cell type marker genes from the CellMarker database (http://biocc.hrbmu.edu.cn/CellMarker/) for cell annotation. A bar chart illustrating cell proportions across samples was generated to represent the distribution of cells within each sample.

    Cell Correlation Analysis

    The relationship between different cells in the PCA space was examined by constructing cluster dendrograms based on PCA dimensions, with the aim of analyzing the Euclidean distances between the cells. Additionally, the correlation among various cell types was evaluated using the average gene expression data.

    GSEA and Gene Set Variation Analysis (GSVA)

    Differential expression analysis was conducted for each cell type, comparing control samples with DN and CKD samples. The log2FoldChange (FC) values for each gene were sorted in descending order for each cell type. GSEA was then performed using the clusterProfiler package (v 3.16.0), with the Kyoto Encyclopedia of Genes and Genomes (KEGG) gene set as the background (|Normalized Enrichment Scores (NES)| > 1, NOM p < 0.05).16 The GSVA package (v 1.46.0) was used to compute GSVA scores across all samples for different cell types based on the h.all.v2022.3.Hs.symbols.gmt gene set.17 The limma package (v 3.54.0) was then applied to assess the statistical significance in pathway differences between control samples and CKD or DN samples (p < 0.05).18

    Cell Communication and Pseudotime Analysis

    CellPhoneDB analysis was performed separately on the control, CKD, and DN samples. The receptor-ligand pairs were filtered with a threshold of p < 0.05 and a minimum mean expression value > 1. Key cells were selected based on annotated cell types according to literature reports.19,20 To examine the differentiation status of key cells at different periods, pseudotime analysis was conducted using Monocle (v 2.14.0), providing insights into the progression of cellular differentiation over time.21

    Identification of Candidate Genes

    To identify candidate genes, differentially expressed genes (DEGs) in key cell types were compared between control and DN samples, and between control and CKD samples. DEGs were selected based on the following criteria: |average log2FC| > 0.25, pct > 0.1, and adj.p < 0.05. The DEGs identified between control and DN samples were designated as DEGs1, while those between control and CKD samples were designated as DEGs2. A Venn diagram tool (http://bioinformatics.psb.ugent.be/webtools/Venn/), was used to find the intersection of DEGs1 and DEGs2, resulting in a list of candidate genes shared by both DN and CKD groups. Gene Ontology (GO) and KEGG pathway analyses were subsequently performed using the clusterProfiler package (v 3.16.0) to investigate the shared functions and pathways of these candidate genes.16

    PPI Network Analysis

    To explore interactions among candidate genes, a PPI network was constructed using the STRING database (https://string-db.org) with a confidence threshold of 0.4. Proteins not connected from the main network were excluded, allowing the focus to remain on the hub genes. The MCODE function in Cytoscape (v 3.10.1) was then used to analyze sub-networks of these hub genes, specifically highlighting the TOP1 sub-network based on the following parameters: degree cutoff = 2, node score cutoff = 0.2, K-core = 2, and max depth = 100).22 The CytoHubba plugin was used to rank hub genes according to four scoring methods (MCC, MNC, Closeness, and Degree). The top 10 genes from each scoring method were selected and genes that consistently appeared across all four methods were designated as biomarkers.

    Enrichment Analysis and Gene Co-Expression Network of Biomarkers

    The correlation coefficients between gene expression in the control versus DN samples and control versus CKD samples were calculated and ranked. This ranking allowed for further GSEA analysis with thresholds set at (|NES| > 1 and adj.p < 0.05). GO and KEGG gene sets were used as background for this analysis. The GeneMANIA database (http://genemania.org) was employed to predict genes that interact with the biomarkers and explore their associated biological functions, facilitating the construction of a gene co-expression network.

    To investigate the activity of upstream pathways associated with the biomarkers, their corresponding pathways were retrieved using the SPEED2 database (https://speed2.sys-bio.net/). The activities of these enriched upstream pathways were quantified using the Bates test and subsequently ranked.

    Biomarker-Drug-Disease Network

    The Comparative Toxicogenomics Database (CTD) (http://ctdbase.org/) was used to predict drugs targeting the biomarkers and to screen relationship pairs in human species. Additionally, the CTD database provided the diseases associated with the identified drugs, which were then visualized in a biomarker-drug-disease network.

    Expression Analysis of Biomarkers

    The “polygenic query” function in GTEx (v 8, https://www.gtexportal.org/home/) was utilized to analyze the expression of biomarkers across different cells and tissues. The expression levels of biomarkers in the annotated cells were assessed, followed by a comparison of expression differences between the control, CKD, and DN groups. Additionally, the expression patterns of biomarkers were analyzed throughout pseudotime based on previous pseudotime analysis results.

    Statistical Analysis

    Data processing and analysis were performed using R software (version 4.2.1). In the bioinformatics analysis, the Wilcoxon rank-sum test was used to examine the differences between the 2 groups. A P-value less than 0.05 was regarded as statistically significant. In addition, the bioinformatics tools and databases used in this study are shown in Supplementary Table 2.

    Results

    Annotation of 10 Cell Types

    The quality of sequencing was assessed, with a Q30 value for all samples exceeding 74%, and the Q20 value surpassing 82% (Supplementary Table 3). Moreover, the sequencing saturation for all samples was greater than 81%, with a mapping rate of over 85% (Supplementary Tables 4 and 5). These results confirm that the sequencing quality of all samples was high, making them suitable for further analysis. After filtering out low-quality cells, the initial cell count of 61,935 was reduced to 28,938 (Supplementary Figure 1). To minimize computational load, the top 2000 most variable genes were selected for PCA (Supplementary Figure 2). The genes from the top nine PCs are shown in Figure 1A, and the top 20 PCs, chosen based on the elbow plot, were used for unsupervised clustering (Figure 1B). A total of 27 clusters were identified, and 10 cell types were annotated: natural killer (NK) T cells, T cells, NK cells, monocytes, B cells, macrophages, mast cells, dendritic cells, and plasma cells (Figure 1C and D, Supplementary Figure 3). T cells, NK T cells, and monocytes were most prevalent across the samples (Figure 1E).

    Figure 1 Continued.

    Figure 1 Annotation of cluster subtypes and unsupervised clustering analysis of single-cell samples. (A and B) PCA analysis and elbow plot for determining the optimal PCs. (C and D) Heatmap of cell clustering based on genes involved in t-SNE dimensionality reduction across the samples. (E) Proportion of cell clusters in control, DN, and CKD samples.

    Abbreviations: PCA, Principal component analysis; PCs, Principal components; t-SNE, t-distributed stochastic neighbor embedding; DN, Diabetic nephropathy; CKD, Chronic kidney disease.

    Pathway Similarities Between CKD and DN Across Cell Types

    The dendrogram showed that cells in close proximity demonstrate higher similarity. Notably, NK cells and NK T cells showed a greater degree of similarity to each other, followed by a closer resemblance to T cells (Figure 2A). A strong positive correlation was also observed between NK cells and NK T cells (Figure 2B). The DEGs in dendritic cells between control and CKD as well as DN samples were enriched in pathways such as the proteasome, endometrial cancer, and apoptosis. In macrophages, the NOD-like receptor signaling pathway was the enriched pathway, while oxidative phosphorylation, non-alcoholic fatty liver disease, and valine, leucine, and isoleucine degradation were the key pathways in mast cells. The enriched pathways for the nine cell types are shown in Supplementary Figure 4 Plasma cells from DN samples were not analyzed due to an insufficient sample size). Significant differences were observed in most pathways between DN and normal samples in T cells, NK T cells, monocytes, B cells, and NK cells. Similarly, notable pathway differences were found between CKD and control samples in T cells, NK T cells, monocytes, and NK cells. The pathway activities in T cells, NK T cells, monocytes, and NK cells were found to be elevated in both CKD and DN, with similar activation patterns suggesting a resemblance between the two conditions (Figure 2C and D).

    Figure 2 CKD and DN samples enriched in pathways of nine cell types. (A) Clustering tree diagram. (B) Heatmap showing cell correlation. (C and D) GSVA analysis of cell subgroups between the control and CKD groups, and the control and DN group groups.

    Abbreviations: CKD, Chronic kidney disease; DN, Diabetic nephropathy; GSVA, Gene set variation analysis.

    Disruption and Imbalance of Cell Communication in DN and CKD

    Cell communication patterns varied markedly between conditions, with a significant increase in cell communication frequency observed in CKD compared to the control group, while the frequency of cell communication was significantly reduced in DN (Figure 3A–F). This indicates that the occurrence of DN and CKD may be associated with disruptions and imbalances in cell communication.

    Figure 3 Disrupted and imbalanced cell communication in DN and CKD. (A, C, E) Heatmaps displaying the relationship between the selected CKD and DN genes and their corresponding expression pathways in the control group, along with changes in gene expression levels. (B, D, F) Cell communication trajectories for control, CKD, and DN samples.

    Abbreviations: DN, Diabetic nephropathy; CKD, Chronic kidney disease.

    Identification of 119 Candidate Genes Associated with Both CKD and DN

    Analysis of myeloid cell subtypes, including monocytes, macrophages, mast cells, and dendritic cells, identified these as key cell types involved in both CKD and DN. Differential expression analysis revealed 297 DEGs (DEGs1) between the control and CKD samples and 277 DEGs (DEGs2) between the control and DN samples in these cell types (Figure 4A and B). Upon intersecting these datasets, 119 candidate genes associated with both CKD and DN were obtained (Figure 4C). The involvement of these candidate genes in disease was linked to several KEGG pathways, such as viral life cycle, measles, malaria, and B cell receptor signaling. Additionally, GO functions related to these genes included negative regulation of MAP kinase activity, toll-like receptor 4 signaling pathway, immunological synapse, platelet alpha granule lumen, amyloid-beta binding, and chemokine receptor binding (Figure 4D and E).

    Figure 4 Screening of key genes. (A and B) Manhattan plots illustrating differentially expressed genes across each chromosome for CKD vs control and DN vs control, respectively (left to right). The y-axis represents -log10(p) values, and the x-axis represents chromosomes, visualizing gene expression across the genome. (C) Intersection of candidate genes relevant to both CKD and DN. (D) KEGG and GO analysis of candidate genes. (E) Bubble plot showing distinct enrichment items, with each node representing a specific biological function. KEGG pathways identified include viral life cycles, viral protein-cytokine receptor interactions, measles, B-cell receptor signaling, African trypanosomiasis, malaria, phagosome, cell adhesion molecules, antigen processing and presentation, and hematopoietic cell lineage.

    Abbreviations: DN, Diabetic nephropathy; CKD, Chronic kidney disease; KEGG, Kyoto Encyclopedia of Genes and Genomes; GO, Gene Ontology.

    MX1, IRF7, STAT1, and ISG15 Were Identified as Biomarkers

    From the initial 119 candidate genes, 21 genes corresponding to discrete proteins were excluded, resulting in a PPI network consisting of 98 proteins (Figure 5A). The top subnetwork identified by the MCODE function included 25 nodes and 267 edges, revealing strong interactions among the proteins (Figure 5B). By intersecting the top 10 genes obtained from the MCC, MNC, Closeness, and Degree scores in the cytoHubba plugin, four biomarkers were identified: MX1, IRF7, STAT1, and ISG15 (Figure 5C). A gene co-expression network was constructed, identifying 20 genes interacting with these biomarkers, primarily involved in functions such as response to type I interferon, cellular response to type I interferon, and viral response (Figure 5D). Additionally, pathway analysis showed that the JAK-STAT, TLR, and TNFa signaling pathways displayed elevated biological activity, while the Hippo and Wnt pathways showed down-regulation in their activities (Figure 5E).

    Figure 5 Associations between key genes and biomarkers in the sample. (A and B) Interaction analysis of 119 candidate genes using a PPI network constructed using STRING (https://string-db.org), with a confidence score of 0.4, identifying 21 discrete proteins and a network comprising 98 interacting proteins. The network contains 98 nodes and 569 edges, visualized using Cytoscape (version 3.10.1). (C) Biomarker identification through MCC, MNC, Closeness, and Degree scores using the cytoHubba plugin, examining the expression activity of four biomarkers across 16 major cell communication signaling pathways. (D) GeneMANIA network analysis, displaying the four biomarker genes in the inner circle, with the outer circle showing other genes related to them. Each gene color denotes its biological pathway, with a high correlation density indicating essential biological functions and significant interactions with other genes. (E) Upstream pathway analysis of the biomarkers.

    Abbreviations: PPI, protein-protein interaction; MNC, Maximum Neighborhood Component; MCC, Maximal Clique Centrality; STRING, Search Tool for the Retrieval of Interacting Genes/Proteins.

    Lysosome Pathway Enrichment of MX1, IRF7, STAT1, and ISG15

    To further explore the functions and pathways associated with the identified biomarkers, GSEA analysis was conducted. The results indicated that MX1 was enriched in pathways such as lysosome, degradation of other glycans, and glutathione metabolism in both CKD and DN (Supplementary Figure 5A and B). IRF7 showed involvement in lysosome-related processes, and was additionally enriched in pathways associated with Vibrio cholerae and Leishmania infections in both CKD and DN (Supplementary Figure 5C and D). STAT1 was linked to lysosome, insulin signaling pathway, Fc gamma R (FcγR)-mediated phagocytosis, and other pathways across CKD and DN (Supplementary Figure 5E and F). Lysosome, systemic lupus erythematosus, glutathione metabolism, and other pathways were enriched by ISG15 in both CKD and DN (Supplementary Figure 5G and H). All biomarkers showed enrichment in the lysosome pathway. Moreover, in CKD, MX1, IRF7, STAT1, and ISG15 were all enriched in the oxidative phosphorylation pathway. In DN, they were all enriched in the FcγR mediated phagocytosis pathway. In addition, a biomarker-drug-disease network was constructed, comprising 165 nodes, including the four biomarkers, 151 drugs (eg, acetylcysteine, acrolein, and alpha-pinene), and 10 diseases (eg, diabetes mellitus, diabetes complications, and diabetic angiopathies), with a total of 635 interaction pairs (Supplementary Figure 6).

    Elevated Expression of MX1 and IRF7 in Dendritic Cells

    The expression levels of the biomarkers MX1, STAT1, and ISG15 were highest in the neuronal cells of the esophagus muscularis, while expression data for IRF7 was unavailable (Figure 6A). The expression levels of the biomarkers in each cell type across different group samples (control, CKD, DN) are shown in Figure 6B. IRF7 showed high expression in dendritic cells in all samples, and MX1 exhibited elevated expression specifically in dendritic cells, particularly in DN and CKD samples (Figure 6C). In contrast, STAT1 and ISG15 were widely expressed in macrophages, monocytes, NK cells, and NK T cells. Notably, these four biomarkers showed significant differential expression in NK cells, T cells, B cells, and NK T cells (Figure 6D–G). The pseudotime analysis highlighted that the significantly elevated expression of MX1 and IRF7 in dendritic cells is associated with myeloid cells differentiation into dendritic cells (Figure 7A–G).

    Figure 6 Connection between expression levels of biomarkers and each cell type. (A) Expression levels of biomarkers across different tissues and cells. (B) Expression levels of biomarkers in individual cell types. (CG) Differential expression of the four biomarkers in each cell type, presented in boxplots. These results show significant differences in the expressions of MX1, STAT1, ISG15, and IRF7 in NK cells, T cells, B cells, and NKT cells. *p<0.05, there is evidence of significant difference; **p<0.01; ***p<0.005; ****p<0.001, with strong evidence of significant difference (P value was used as the standard for figure (DG) screening of differentially expressed genes).

    Abbreviations: ns, no significant difference.

    Figure 7 Pseudotime analysis showing the differentiation states of cells. Branch points indicate potential decision points in cellular processes. (AC) Each point represents a cell, with cells exhibiting similar cellular states grouped together. Branch points in the pseudotime trajectory indicate potential decision points in the cell’s biological process (eg, four branch points in this analysis). Cells are color-labeled according to pseudotime, state, and group. Integrating biomarker expression data, the differentiation states of genes related to disease progression can be identified. (DG) Pseudotime analysis of biomarkers. The results reveal that MX1 and IRF7 genes show significantly higher expression levels in dendritic cells, suggesting that MX1 and IRF7 may play a role in the differentiation of myeloid cells into dendritic cells. In contrast, the other two biomarkers (STAT1 and ISG15) did not exhibit significant differences in expression, indicating they may have a lesser impact on this differentiation process.

    Discussion

    T2DM and CKD are both widespread chronic diseases. The most common microvascular complication of T2DM is DN, which is the leading cause of CKD.23 Currently, the diagnosis of these two conditions relies on traditional markers such as estimated glomerular filtration rate (eGFR), urinary albumin measurement, and creatinine levels, especially in the absence of renal biopsy.24 Early diagnosis of CKD and DN is often subjective due to the lack of non-invasive biomarkers.25 This limitation complicates the design of clinical trials, impeding efforts to identify effective treatments, facilitate early detection, and ensure timely diagnosis. Additionally, reducing cardiovascular mortality and slowing the progression to ESRD remain significant unmet medical needs for patients with CKD and DN.26

    This study offers new insights into the molecular mechanisms underlying DN and CKD, by identifying common biomarkers and exploring the biological processes involved. Using scRNA-seq technology, we analyzed cell-specific gene expression changes across control, CKD, and DN groups, identifying differentially expressed genes within distinct cell subpopulations. Through further examination of the signaling pathways within these cell clusters, this study provides a theoretical foundation for understanding biomarkers related to the progression of CKD and DN, their roles in immune response, and their potential as therapeutic targets.

    In this study, four biomarkers—MX1, IRF7, STAT1, and ISG15—were identified through differential expression analysis and the PPI network analysis, showing notable expression in both DN and CKD samples.

    Interferon-stimulated gene 15 (ISG15) is a 15kD ubiquitin-like protein induced by the binding of interferon-α (IFN-α) to the promoters of interferon (IFN) regulatory factor (IRF) and the interferon-stimulated response element. ISG15 has been implicated in several intracellular processes, including autophagy, apoptosis, and signal transduction. As a cytokine, it activates Janus kinase (JAK) and the JAK/STAT signaling pathway, which mediates various physiological and pathological responses, such as cell proliferation, differentiation, apoptosis, and immune regulation.27 Recent studies indicate that JAK/STAT pathway activation exacerbates renal fibrosis and glomerulosclerosis, while ISG15 overexpression contributes to systemic inflammation and CKD.28,29 At the same time, ISG15 can modify viral proteins or host proteins, thereby inhibiting viral replication.30 ISG15 modification can directly interfere with the life cycle of the virus.31 And the resistance of ISG15-deficient cells to paramyxovirus is reduced, indicating their direct antiviral activity.32 In oral squamous cell carcinoma, tumor cell-derived ISG15 promotes fibroblast recruitment, promoting tumor growth and metastasis through CD11a-dependent glycolytic reprogramming.33 In pancreatic and renal clear cell carcinoma, ISG15 levels are elevated and high expression is associated with adverse clinical outcomes.34,35 ISG15 can also promote tumor cell migration and immunosuppression by inducing the macrophage M2-like phenotype.36 To sum up, ISG15 has shown a pleiotropic effect in antiviral immunity, tumor progression and metastasis, and tumor microenvironment regulation. Its complex functional mechanism in different pathophysiological processes provides rich and promising research directions for the development of prevention and treatment strategies for related diseases.

    This study found that MX1 was specifically highly expressed in DN and CKD samples. The MX1 gene encodes an interferon-induced protein that is involved in the cell’s antiviral immune response.37 In the context of diabetes, persistent hyperglycemia and lipid metabolism disorders may activate MX1, leading to chronic inflammation, which may promote pathological changes in DN.38 In CKD, MX1 may reduce the inflammatory response caused by viral infection by inhibiting viral replication, thereby producing a protective effect on CKD.39,40 Additionally, the degree of methylation of the MX1 gene promoter is correlated with the severity of COVID-19 and there may be gender differences.41 This suggests that the expression level of MX1 can be used as an early indicator of viral infection, and its gene polymorphism is also related to the risk of autoimmune disease, and is of great value in the judgment of infection type, individualized treatment and disease risk assessment.

    Signal transducer and activator of transcription 1 (STAT1) is a cytoplasmic transcription factor activated by various stimuli, regulating the human immune system.42 Moreover, STAT1 mediates interferon signaling pathways and plays a crucial role in antiviral (eg, HBV, HCV, HIV) and antibacterial (eg, Mycobacterium tuberculosis) immune responses. Its phosphorylation levels reflect the progression of infection.43–45 Additionally, STAT1 gain-of-function mutations are associated with chronic mucosal skin candidiasis and systemic lupus erythematosus (SLE), while loss-of-function mutations lead to severe immunodeficiency.46,47 STAT1 exhibits a dual role in cancer: high expression in prostate and breast cancers may indicate a better prognosis, but it may also promote immune escape in some solid tumors.48 These findings suggest that STAT1 could serve as a monitoring indicator for infectious disease progression, a molecular diagnostic marker for autoimmune diseases, and a potential target for tumor prognosis evaluation. Regulating STAT1 may provide novel strategies for precise treatment of related diseases.

    IRF7 (Interferon Regulatory Factor 7) is a key member of the IRF family, playing a pivotal role in innate immunity and antiviral responses.49,50 Studies have shown that IRF7 expression correlates with disease activity in SLE patients, with elevated mRNA levels positively correlated with serum IFN levels, IFN scores, and SLEDAI scores.51,52 In acute myeloid leukemia, inhibiting TOX exerts anti-tumor effects by upregulating IRF7 expression.53 Furthermore, IRF7 mediates the transcription of MCP-1, an obesity-related molecule.54 These findings indicate that changes in IRF7 expression are linked to disease development and may serve as a potential biomarker for diagnosis.

    MX1, IRF7, STAT1, and ISG15 are core regulatory molecules in the type I interferon (IFN-α/β) response pathway, interacting with each other in complex ways. STAT1, as a central signal node, is phosphorylated by JAK kinase under IFN-γ or IFN-α/β stimulation. This results in the formation of a dimer that translocates to the nucleus and directly activates IRF7 transcription.55 IRF7, in turn, amplifies IFN-α/β production, creating a positive feedback loop, and induces MX1 and ISG15 expression.56 MX1, an antiviral effector protein, relies on the STAT1-IRF7 axis, while ISG15 stabilizes STAT1 and IRF7 proteins via ubiquitination (ISGylation) and enhances their transcriptional activity.57 Additionally, ISG15 regulates MX1 oligomerization through non-covalent binding, impacting its antiviral function.58 In CKD and DN, these complex interactions may lead to the oversecretion of pro-inflammatory factors (eg, TNF-α, IL-6) and dysregulated cytotoxic immune responses, accelerating tissue damage.

    Furthermore, exploring the relationship between MX1, IRF7, STAT1, and ISG15 and traditional renal function markers (eg, serum creatinine, urea nitrogen, and urine protein) is valuable. Serum creatinine levels are influenced by muscle metabolism and glomerular filtration capacity,59 urea nitrogen reflects protein metabolism and renal excretion function,60 and urinary protein indicates impaired glomerular filtration barrier.61 In contrast, MX1, IRF7, STAT1, and ISG15 are more involved in immunomodulation and inflammatory response pathways.62,63 In kidney disease progression, local inflammation in the kidney and immune cell activation64,65 can alter the expression of these biomarkers. In early CKD, the immune-inflammatory response begins subtly, and the expression of MX1, IRF7, etc., may change before traditional markers like serum creatinine or urea nitrogen significantly deviate. At this stage, urine protein may also remain at critical levels.66–69 This suggests that biomarkers such as MX1 could serve as early warning indicators, complementing traditional markers. As the disease progresses and traditional markers become more abnormal, these novel biomarkers may increase, providing a more comprehensive basis for disease assessment.

    GSEA results indicated that all four biomarkers were enriched in the Oxidative Phosphorylation pathway in CKD, while in DN, they were enriched in the FcγR-mediated Phagocytosis pathway. Kidney cells typically rely on Oxidative Phosphorylation to maintain physiological functions like reabsorption and secretion, processes requiring substantial energy.70 However, in CKD, kidney cell metabolic pathways are altered, and their dependence on Oxidative Phosphorylation is enhanced.71 Oxidative Phosphorylation is a major source of intracellular reactive oxygen species (ROS). In CKD, metabolic disturbances and inflammatory responses may overactivate this pathway, resulting in excessive ROS production.71 Excessive ROS can damage cellular proteins, lipids, and DNA, triggering oxidative stress and accelerating renal fibrosis and functional decline.72 FcγR-mediated phagocytosis is crucial for immune function, facilitating the uptake and clearance of phagocytes (eg, macrophages, neutrophils) that recognize target cells or granules bound to antibodies.73 In early DN, immune complexes may accumulate in renal tissues, activating the complement system and inflammatory responses, thereby exacerbating kidney damage.74 If phagocytosis fails to clear target cells completely, residual antigens may continue to stimulate the immune response, leading to chronic inflammation and renal fibrosis.75 These findings highlight the differences in enriched pathways between CKD and DN, shedding light on the distinct pathogenesis and progression of each disease. This insight is crucial for understanding disease mechanisms and developing targeted therapeutic strategies.

    This study found that IRF7 expression was elevated in dendritic cells (DCs). In CKD and DN, immune cells like DCs are continuously activated during disease progression.76 Plasmacytoid DCs (pDCs) are the primary producers of IFN-I,77 and in pDCs, IRF7 expression is regulated by NFATC3, which enhances IFN production.78 Upon stimulation by TLR7 or TLR9, IRF7 is activated, promoting IFN-α secretion.79 High IRF7 expression in DCs strengthens their antiviral immune response,50,80 enhances antigen presentation, and regulates the Th1 immune response and inflammatory factor release.81–84 In CKD and DN, abnormal IRF7 expression may contribute to disease progression via two mechanisms: excessive activation can cause persistent inflammation, macrophage infiltration, and proinflammatory factor release (eg, TNF-α, IL-6), exacerbating fibrosis;38,85,86 additionally, IRF7 may worsen podocyte damage and glomerular basement membrane thickening under metabolic stress (eg, high sugar or glycosylation end product stimulation).87 Notably, the high-sugar environment in DN amplifies IRF7-mediated inflammation, creating a vicious “metabolic-inflammatory” cycle.88 Thus, understanding IRF7’s role in DCs provides key insights into the pathogenesis of CKD and DN, offering future directions for developing targeted therapies to block disease progression.74–77

    Our study also highlights that STAT1 and ISG15 are widely expressed in macrophages, monocytes, NK cells, and NKT cells. In macrophages, IFN-γ stimulates the phosphorylation of STAT1 by JAK1/JAK2, promoting STAT1 dimerization, nuclear translocation, and the expression of pro-inflammatory genes such as IRF1 and CXCL10, enhancing M1 polarization and antibacterial function.89,90 ISG15 regulates cytokine expression (eg, TNF-α, IL-6) by activating the JAK-STAT pathway, modulating immune response intensity.91 In monocytes, STAT1 enhances the inflammatory response via the TLR/MyD88 pathway,92 while ISG15 may regulate cell migration and phagocytosis through NF-κB signaling.58,93 For NK and NKT cells, STAT1 mediates the IFN-γ feedback loop, promoting granzyme and perforin expression,94,95 while ISG15 supports IFN-γ production and cytotoxic function by stabilizing STAT1/STAT4.56,96 Free ISG15 also enhances NK cell cytotoxicity via LFA-1 receptors.57 Importantly, STAT1 can compete with STAT3 for DNA binding to regulate immune balance, but over-activation may lead to chronic inflammation and is associated with autoimmune and metabolic diseases like diabetic nephropathy.55,97 These findings underscore the core role of STAT1 and ISG15 in the innate immune system, offering new insights into immune cell activation and laying the foundation for targeted therapies in inflammatory and metabolic diseases.90

    Furthermore, sodium-glucose cotransporter 2 inhibitors have been shown to interfere with the polarization of DCs by reducing receptor pairing between M2 macrophages and T cells. In this study, cell communication analysis of DN and CKD groups revealed that B cells, NK cells, T cells, and monocytes exhibited the closest interactions. The four biomarkers, MX1, IRF7, STAT1, and ISG15, were widely expressed in these cell populations. These findings suggest that these biomarkers play central roles in the immune response and the progression of CKD and DN. These biomarkers could serve as valuable targets for predicting disease progression.

    However, this study has certain limitations. First, single-cell sequencing technology presents challenges such as high costs and time consumption, making it difficult to fully assess the accuracy of these markers in disease evaluation. More importantly, the specific functional mechanisms, clinical translational potential, and diagnostic value of these biomarkers require systematic validation in independent cohorts. Additionally, the dynamic changes in relevant signaling pathways during CKD and DN progression, their correlation with disease stage and severity, and the clinical application value of these markers still need further investigation. To address these limitations, future research will focus on enhancing the clinical applicability of single-cell sequencing, including cost reduction, time efficiency, and improved accuracy and reliability. Interdisciplinary collaboration will be promoted to facilitate its clinical use. We plan to explore the impact of biomarkers on disease-related cell behavior and physiological processes through cell culture and animal models (eg, PDO, PDX). This will involve verifying whether biomarker regulation can reverse disease phenotypes and elucidate underlying mechanisms. Concurrently, large-scale, multicenter clinical samples will be collected to investigate the MX1/IRF7/STAT1/ISG15 pathway using gene-editing technologies. Furthermore, we will compare biomarker expression across CKD, DN, and other renal diseases to assess specificity. In terms of clinical translation, we will collaborate with the Clinical Center for Nephrology to examine the correlation between biomarkers, disease staging, and treatment response. Clinical measurement techniques such as ELISA, mass spectrometry, and immunohistochemistry will be used to correlate biomarker expression with clinical data. Samples from different geographic regions will be collected to assess the generalizability of our findings.

    Conclusion

    In this study, through single-cell RNA sequencing and the application of a series of bioinformatics methods, four biomarkers (MX1, IRF7, STAT1, ISG15) in CKD and DN were identified. During the clinical diagnosis process, detecting the expression levels of biomarkers in patients may serve as a means of auxiliary diagnosis for CKD and DN, and also as an important basis for predicting disease progression. Meanwhile, in the clinical treatment of CKD and DN, these biomarkers can be considered as therapeutic targets.

    Data Sharing Statement

    All data generated or analysed during this study are included in this article. Further enquiries can be directed to the corresponding author.

    Ethics Approval and Consent to Participate

    This study was conducted with approval from the Ethics Committee of The Second Hospital Affiliated to Kunming Medical University (PJ-2021-36). This study was conducted in accordance with the declaration of Helsinki. Written informed consent was obtained from all participants.

    Acknowledgments

    We would like to acknowledge the hard and dedicated work of all the staff that implemented the intervention and evaluation components of the study.

    Funding

    Yunnan Revitalization Talent Support Program (Youth talent project: NO.YNWR-QNBJ-2020-269) and (Famous doctors project: NO.YNWR-MY-2019-075). Reserve Talents Project for Young and Middle-aged Academic and Technical leaders in Yunnan Province (202005AC160024). Yunnan Fundamental Research Kunming Medical University Joint Projects (grant NO. 202201AY070001-101). National Clinical Research Center of Chronic Kidney Disease, the Second Affiliated Hospital of Kunming Medical University.(Project Number: GF2020003). The Second Affiliated Hospital of Kunming Medical University talent echelon cultivation project-Academic leader (RCTDXS-202303).

    Disclosure

    The authors declare that they have no competing interests.

    References

    1. Ammirati AL. Chronic kidney disease. Rev Assoc Med Bras. 2020;66Suppl 1(Suppl 1):s03–s09. PMID: 31939529. doi:10.1590/1806-9282.66.S1.3

    2. McGrath K, Edi R. Diabetic kidney disease: diagnosis, treatment, and prevention. Am Fam Physician. 2019;99(12):751–759.

    3. Jun Z, Xiaoying L, Min L. Analysis of correlation between heart rate variability and vascular endothelial injury and microinflammation in patients with chronic kidney disease. Chinese Medical Engineerin. 2024;32(05):85–88. doi:10.19338/j.issn.1672-2019.2024.05.019

    4. Stevens PE, Levin A; Kidney Disease: Improving Global Outcomes Chronic Kidney Disease Guideline Development Work Group Members. Evaluation and management of chronic kidney disease: synopsis of the kidney disease: improving global outcomes 2012 clinical practice guideline. Ann Intern Med. 2013;158(11):825–830. doi:10.7326/0003-4819-158-11-201306040-00007

    5. Yamanouchi M, Furuichi K, Hoshino J, Ubara Y, Wada T. Nonproteinuric diabetic kidney disease. Clin Exp Nephrol. 2020;24(7):573–581. doi:10.1007/s10157-020-01881-0

    6. Chen TK, Hoenig MP, Nitsch D, et al. Advances in the management of chronic kidney disease. BMJ. 2023;383:e074216. doi:10.1136/bmj-2022-074216

    7. Hou JH, Zhu HX, Zhou ML, et al. Changes in the spectrum of kidney diseases: an analysis of 40,759 biopsy-proven cases from 2003 to 2014 in China. Kidney Dis. 2018;4(1):10–19. doi:10.1159/000484717

    8. Komici K, Femminella GD, de Lucia C, et al. Predisposing factors to heart failure in diabetic nephropathy: a look at the sympathetic nervous system hyperactivity. Aging Clin Exp Res. 2019;31(3):321–330. doi:10.1007/s40520-018-0973-2

    9. Berest I, Tangherloni A. Integration of scATAC-Seq with scRNA-Seq data. Methods Mol Biol. 2023;2584:293–310. doi:10.1007/978-1-0716-2756-3_15

    10. Macosko EZ, Basu A, Satija R, et al. Highly parallel genome-wide expression profiling of individual cells using nanoliter droplets. Cell. 2015;161(5):1202–1214. doi:10.1016/j.cell.2015.05.002

    11. Kidney Disease: Improving Global Outcomes (KDIGO) Diabetes Work Group. KDIGO 2022 clinical practice guideline for diabetes management in chronic kidney disease. Kidney Int. 2022;102(5S):S1–S127. doi:10.1016/j.kint.2022.06.008

    12. Kidney Disease: Improving Global Outcomes (KDIGO) Diabetes Work Group. KDIGO 2020 clinical practice guideline for diabetes management in chronic kidney disease. Kidney Int. 2020;98(4S):S1–S115. doi:10.1016/j.kint.2020.06.019

    13. Liao Y, Raghu D, Pal B, Mielke LA, Shi W. cellCounts: an R function for quantifying 10x Chromium single-cell RNA sequencing data. Bioinformatics. 2023;39(7):btad439. doi:10.1093/bioinformatics/btad439

    14. Hao Y, Hao S, Andersen-Nissen E, et al. Integrated analysis of multimodal single-cell data. Cell. 2021;184(13):3573–3587.e29. doi:10.1016/j.cell.2021.04.048

    15. Germain PL, Lun A, Garcia Meixide C, Macnair W, Robinson MD. Doublet identification in single-cell sequencing data using scDblFinder. F1000Res. 2021;10:979. doi:10.12688/f1000research.73600.1

    16. Wu T, Hu E, Xu S, et al. clusterProfiler 4.0: a universal enrichment tool for interpreting omics data. Innovation. 2021;2(3):100141. doi:10.1016/j.xinn.2021.100141

    17. Hänzelmann S, Castelo R, Guinney J. GSVA: gene set variation analysis for microarray and RNA-seq data. BMC Bioinf. 2013;14(1):7. doi:10.1186/1471-2105-14-7

    18. Ritchie ME, Phipson B, Wu D, et al. limma powers differential expression analyses for RNA-sequencing and microarray studies. Nucleic Acids Res. 2015;43(7):e47. doi:10.1093/nar/gkv007

    19. Conway BR, O’Sullivan ED, Cairns C, et al. Kidney single-cell atlas reveals myeloid heterogeneity in progression and regression of kidney disease. J Am Soc Nephrol. 2020;31(12):2833–2854. doi:10.1681/ASN.2020060806

    20. Bell RMB, Denby L. Myeloid heterogeneity in kidney disease as revealed through single-cell RNA sequencing. Kidney360. 2021;2(11):1844–1851. doi:10.34067/KID.0003682021

    21. Qiu X, Mao Q, Tang Y, et al. Reversed graph embedding resolves complex single-cell trajectories. Nat Methods. 2017;14(10):979–982. doi:10.1038/nmeth.4402

    22. Shannon P, Markiel A, Ozier O, et al. Cytoscape: a software environment for integrated models of biomolecular interaction networks. Genome Res. 2003;13(11):2498–2504. doi:10.1101/gr.1239303

    23. Rayego-Mateos S, Rodrigues-Diez RR, Fernandez-Fernandez B, et al. Targeting inflammation to treat diabetic kidney disease: the road to 2030. Kidney Int. 2023;103(2):282–296. doi:10.1016/j.kint.2022.10.030

    24. Aldrich S, Ashjian E. Use of GLP-1 receptor agonists in patients with T2DM and chronic kidney disease. Nurse Pract. 2019;44(3):20–28. doi:10.1097/01.NPR.0000553396.65976.bb

    25. Anders HJ, Huber TB, Isermann B, Schiffer M. CKD in diabetes: diabetic kidney disease versus nondiabetic kidney disease. Nat Rev Nephrol. 2018;14(6):361–377. doi:10.1038/s41581-018-0001-y

    26. Doshi SM, Friedman AN. Diagnosis and management of type 2 diabetic kidney disease. Clin J Am Soc Nephrol. 2017;12(8):1366–1373. doi:10.2215/CJN.11111016

    27. Mirzalieva O, Juncker M, Schwartzenburg J, Desai S. ISG15 and ISGylation in human diseases. Cells. 2022;11(3):538. doi:10.3390/cells11030538

    28. Jia J, Xu LH, Deng C, et al. Hederagenin ameliorates renal fibrosis in chronic kidney disease through blocking ISG15 regulated JAK/STAT signaling. Int Immunopharmacol. 2023;118:110122. doi:10.1016/j.intimp.2023.110122

    29. He T, Xia Y, Yang J. Systemic inflammation and chronic kidney disease in a patient due to the RNASEH2B defect. Pediatr Rheumatol Online J. 2021;19(1):9. doi:10.1186/s12969-021-00497-2

    30. Kespohl M, Bredow C, Klingel K, et al. Protein modification with ISG15 blocks coxsackievirus pathology by antiviral and metabolic reprogramming. Sci Adv. 2020;6(11):eaay1109. doi:10.1126/sciadv.aay1109

    31. Freitas BT, Scholte FEM, Bergeron É, Pegan SD. How ISG15 combats viral infection. Virus Res. 2020;286:198036. doi:10.1016/j.virusres.2020.198036

    32. Holthaus D, Vasou A, Bamford CGG, et al. Direct antiviral activity of IFN-stimulated genes is responsible for resistance to paramyxoviruses in ISG15-deficient cells. J Immunol. 2020;205(1):261–271. doi:10.4049/jimmunol.1901472

    33. Wang SH, Chen YL, Huang SH, et al. Tumor cell-derived ISG15 promotes fibroblast recruitment in oral squamous cell carcinoma via CD11a-dependent glycolytic reprogramming. Oncogenesis. 2025;14(1):6. doi:10.1038/s41389-025-00549-2

    34. Meng Y, Bian L, Zhang M, et al. ISG15 promotes progression and gemcitabine resistance of pancreatic cancer cells through ATG7. Int J Biol Sci. 2024;20(4):1180–1193. doi:10.7150/ijbs.85424

    35. Xie W, Zhang Y, Zhang Z, Li Q, Tao L, Zhang R. ISG15 promotes tumor progression via IL6/JAK2/STAT3 signaling pathway in ccRCC. Clin Exp Med. 2024;24(1):140. doi:10.1007/s10238-024-01414-z

    36. Chen RH, Xiao ZW, Yan XQ, et al. Tumor cell-secreted ISG15 promotes tumor cell migration and immune suppression by inducing the macrophage M2-like phenotype. Front Immunol. 2020;11:594775. doi:10.3389/fimmu.2020.594775

    37. Wang G, Hua R, Chen X, et al. MX1 and UBE2L6 are potential metaflammation gene targets in both diabetes and atherosclerosis. PeerJ. 2024;12:e16975. doi:10.7717/peerj.16975

    38. Matoba K, Takeda Y, Nagai Y, Kawanami D, Utsunomiya K, Nishimura R. Unraveling the role of inflammation in the pathogenesis of diabetic kidney disease. Int J Mol Sci. 2019;20(14):3393. doi:10.3390/ijms20143393

    39. Salomon R, Staeheli P, Kochs G, et al. Mx1 gene protects mice against the highly lethal human H5N1 influenza virus. Cell Cycle. 2007;6(19):2417–2421. doi:10.4161/cc.6.19.4779

    40. Jakhotia S, Kavvuri R, Raviraj S, Baishya S, Pasupulati AK, Reddy GB. Obesity-related glomerulopathy is associated with elevated WT1 expression in podocytes. Int J Obes. 2024;48(8):1080–1091. doi:10.1038/s41366-024-01509-3

    41. Ghoreshi ZA, Abbasi-Jorjandi M, Asadikaram G, Sharifzak M, Rezazadeh-Jabalbarzi M, Rashidinejad H. Evaluation of MX1 gene promoter methylation in different severities of COVID-19 considering patient gender. Clin Lab. 2022;68(10). doi:10.7754/Clin.Lab.2022.220104

    42. Spitaels J, Van Hoecke L, Roose K, Kochs G, Saelens X. Mx1 in hematopoietic cells protects against thogoto virus infection. J Virol. 2019;93(15):e00193–19. doi:10.1128/JVI.00193-19

    43. Jung SR, Ashhurst TM, West PK, et al. Contribution of STAT1 to innate and adaptive immunity during type I interferon-mediated lethal virus infection. PLoS Pathog. 2020;16(4):e1008525. doi:10.1371/journal.ppat.1008525

    44. Lei WT, Lo YF, Tsumura M, et al. Immunophenotyping and therapeutic insights from chronic mucocutaneous candidiasis cases with STAT1 gain-of-function mutations. J Clin Immunol. 2024;44(8):184. doi:10.1007/s10875-024-01776-9

    45. Ivashkiv LB, Donlin LT. Regulation of type I interferon responses. Nat Rev Immunol. 2014;14(1):36–49. PMID: 24362405. doi:10.1038/nri3581

    46. Kristensen IA, Veirum JE, Møller BK, Christiansen M. Novel STAT1 alleles in a patient with impaired resistance to mycobacteria. J Clin Immunol. 2011;31(2):265–271. PMID: 21057861. doi:10.1007/s10875-010-9480-8

    47. Uzel G, Sampaio EP, Lawrence MG, et al. Dominant gain-of-function STAT1 mutations in FOXP3 wild-type immune dysregulation-polyendocrinopathy-enteropathy-X-linked-like syndrome. J Allergy Clin Immunol. 2013;131(6):1611–1623. doi:10.1016/j.jaci.2012.11.054

    48. Khodarev NN, Beckett M, Labay E, Darga T, Roizman B, Weichselbaum RR. STAT1 is overexpressed in tumors selected for radioresistance and confers protection from radiation in transduced sensitive cells. Proc Natl Acad Sci U S A. 2004;101(6):1714–1719. doi:10.1073/pnas.0308102100

    49. Qing F, Liu Z. Interferon regulatory factor 7 in inflammation, cancer and infection. Front Immunol. 2023;14:1190841. doi:10.3389/fimmu.2023.1190841

    50. Ma W, Huang G, Wang Z, Wang L, Gao Q. IRF7: role and regulation in immunity and autoimmunity. Front Immunol. 2023;14:1236923. doi:10.3389/fimmu.2023.1236923

    51. Renaudineau Y, Charras A, Natoli V, et al. UK jSLE cohort study. Type I interferon associated epistasis may contribute to early disease-onset and high disease activity in juvenile-onset lupus. Clin Immunol. 2024;262:110194. doi:10.1016/j.clim.2024.110194

    52. Xu WD, Zhang YJ, Xu K, et al. IRF7, a functional factor associates with systemic lupus erythematosus. Cytokine. 2012;58(3):317–320. PMID: 22455868. doi:10.1016/j.cyto.2012.03.003

    53. Huang S, Chen Z, Zhong S, et al. Inhibition of TOX exerts anti-tumor effects in acute myeloid leukemia by upregulating IRF7 expression. Eur J Pharmacol. 2025;987:177163. doi:10.1016/j.ejphar.2024.177163

    54. Kuroda M, Nishiguchi M, Ugawa N, et al. Interferon regulatory factor 7 mediates obesity-associated MCP-1 transcription. PLoS One. 2020;15(5):e0233390. doi:10.1371/journal.pone.0233390

    55. Seffens A, Herrera A, Tegla C, et al. STAT3 dysregulation in mature T and NK cell lymphomas. Cancers. 2019;11(11):1711. doi:10.3390/cancers11111711

    56. Swaim CD, Canadeo LA, Monte KJ, Khanna S, Lenschow DJ, Huibregtse JM. Modulation of extracellular ISG15 signaling by pathogens and viral effector proteins. Cell Rep. 2020;31(11):107772. doi:10.1016/j.celrep.2020.107772

    57. Swaim CD, Scott AF, Canadeo LA, Huibregtse JM. Extracellular ISG15 signals cytokine secretion through the LFA-1 integrin receptor. Mol Cell. 2017;68(3):581–590.e5. doi:10.1016/j.molcel.2017.10.003

    58. Nowak K, Jabłońska E, Ratajczak-Wrona W. NF-κB-an important player in xenoestrogen signaling in immune cells. Cells. 2021;10(7):1799. doi:10.3390/cells10071799

    59. Butt B, Ghulam B, Bashir Z, et al. Enhanced creatinine level in diabetic patients maximizing the possibilities of nephropathy and its association with blood urea nitrogen and glomerular filtration rate. Cureus. 2024;16(9):e70482. doi:10.7759/cureus.70482

    60. Ahmad S, Khan MA, Ali R. Blood urea nitrogen (BUN) levels in renal failure: unraveling the complex interplay of protein metabolism and kidney health. Professional Med J. 2024;31(3):7908. doi:10.29309/TPMJ/2024.31.03.7908

    61. Lee SW, Baek SH, Paik JH, et al. Tubular B7-1 expression parallels proteinuria levels, but not clinical outcomes in adult minimal change disease patients. Sci Rep. 2017;7:41859. PMID: 28150736; PMCID: PMC5288792. doi:10.1038/srep41859

    62. Cui N, Liu C, Tang X, et al. ISG15 accelerates acute kidney injury and the subsequent AKI-to-CKD transition by promoting TGFβR1 ISGylation. Theranostics. 2024;14(11):4536–4553. doi:10.7150/thno.95796

    63. Da G, Wang J, Shang J, et al. Nuclear PCGF3 inhibits the antiviral immune response by suppressing the interferon-stimulated gene. Cell Death Discov. 2024;10(1):429. doi:10.1038/s41420-024-02194-x

    64. Cantero-Navarro E, Rayego-Mateos S, Orejudo M, et al. Role of macrophages and related cytokines in kidney disease. Front Med. 2021;8:688060. doi:10.3389/fmed.2021.688060

    65. Imig JD, Ryan MJ. Immune and inflammatory role in renal disease. Compr Physiol. 2013;3(2):957–976. doi:10.1002/cphy.c120028

    66. Shimizu Y, Yasuda S, Kimura T, et al. Interferon-inducible Mx1 protein is highly expressed in renal tissues from treatment-naïve lupus nephritis, but not in those under immunosuppressive treatment. Mod Rheumatol. 2018;28(4):661–669. doi:10.1080/14397595.2017.1404711

    67. Smith JD, Doe JM. The impact of dialysis on patient quality of life. Open J Nephrol. 2021;11(3):31. doi:10.4236/ojneph.2021.113031

    68. Fu Y, Xiang Y, Wang Y, et al. The STAT1/HMGB1/NF-κB pathway in chronic inflammation and kidney injury after cisplatin exposure. Theranostics. 2023;13(9):2757–2773. PMID: 37284446; PMCID: PMC10240827. doi:10.7150/thno.81406

    69. Zheng C, Shang F, Cheng R, Bai Y. STAT1 aggravates kidney injury by NOD-like receptor (NLRP3) signaling in MRL-lpr mice. J Mol Histol. 2024;55(4):555–566. PMID: 38856930. doi:10.1007/s10735-024-10208-2

    70. Johnson AR, Smith LK. Advances in mitochondrial dynamics and cellular energy regulation. Mitochondrial Commun. 2024;3(3):Article100234. doi:10.1016/j.mitoco.2024.03.002

    71. Shi J, Yang Y, Wang YN, et al. Oxidative phosphorylation promotes vascular calcification in chronic kidney disease. Cell Death Dis. 2022;13(3):229. doi:10.1038/s41419-022-04679-y

    72. Mapuskar KA, Vasquez-Martinez G, Mayoral-Andrade G, Tomanek-Chalkley A, Zepeda-Orozco D, Allen BG. Mitochondrial oxidative metabolism: an emerging therapeutic target to improve CKD outcomes. Biomedicines. 2023;11(6):1573. doi:10.3390/biomedicines11061573

    73. Cachofeiro V, Goicochea M, de Vinuesa SG, Oubiña P, Lahera V, Luño J. Oxidative stress and inflammation, a link between chronic kidney disease and cardiovascular disease. Kidney Int Suppl. 2008;111:S4–9. doi:10.1038/ki.2008.516

    74. Acharya D, Li XRL, Heineman RE, Harrison RE. Complement receptor-mediated phagocytosis induces proinflammatory cytokine production in murine macrophages. Front Immunol. 2020;10:3049. doi:10.3389/fimmu.2019.03049

    75. Restrepo BI, Twahirwa M, Rahbar MH, Schlesinger LS. Phagocytosis via complement or Fc-gamma receptors is compromised in monocytes from type 2 diabetes patients with chronic hyperglycemia. PLoS One. 2014;9(3):e92977. doi:10.1371/journal.pone.0092977

    76. Ferracini M, Martins JO, Campos MR, Anger DB, Jancar S. Impaired phagocytosis by alveolar macrophages from diabetic rats is related to the deficient coupling of LTs to the Fc gamma R signaling cascade. Mol Immunol. 2010;47(11–12):1974–1980. doi:10.1016/j.molimm.2010.04.018

    77. Jia Y, Xu H, Yu Q, Tan L, Xiong Z. Identification and verification of vascular cell adhesion protein 1 as an immune-related hub gene associated with the tubulointerstitial injury in diabetic kidney disease. Bioengineered. 2021;12(1):6655–6673. doi:10.1080/21655979.2021.1976540

    78. Wang M, Zhang Y, Zhai Y, Li H, Xie Z, Wen C. The mechanism of Langchuangding in treatment of systemic lupus erythematosus via modulating TLR7-IRF7-IFNα pathway. Heliyon. 2024;10(5):e26022. doi:10.1016/j.heliyon.2024.e26022

    79. Bao M, Wang Y, Liu Y, et al. NFATC3 promotes IRF7 transcriptional activity in plasmacy–toid dendritic cells. J Exp Med. 2016;213(11):2383–2398. doi:10.1084/jem.20160438

    80. Di Domizio J, Cao W. Fueling autoimmunity: type I interferon in autoimmune diseases. Expert Rev Clin Immunol. 2013;9(3):201–210. doi:10.1586/eci.12.106

    81. Owens BM, Moore JW, Kaye PM. IRF7 regulates TLR2-mediated activation of splenic CD11c(hi) dendritic cells. PLoS One. 2012;7(7):e41050. doi:10.1371/journal.pone.0041050

    82. Kumar S, Jeong Y, Ashraf MU, Bae YS. Dendritic cell-mediated Th2 immunity and immune disorders. Int J Mol Sci. 2019;20(9):2159. doi:10.3390/ijms20092159

    83. Di Sabatino A, Pickard KM, Gordon JN, et al. Evidence for the role of interferon-alfa production by dendritic cells in the Th1 response in celiac disease. Gastroenterology. 2007;133(4):1175–1187. doi:10.1053/j.gastro.2007.08.018

    84. Terhune J, Berk E, Czerniecki BJ. Dendritic cell-induced Th1 and Th17 cell differentiation for cancer therapy. Vaccines. 2013;1(4):527–549. doi:10.3390/vaccines1040527

    85. Hung PH, Hsu YC, Chen TH, Lin CL. Recent advances in diabetic kidney diseases: from kidney injury to kidney fibrosis. Int J Mol Sci. 2021;22(21):11857. doi:10.3390/ijms222111857

    86. Pichler R, Afkarian M, Dieter BP, Tuttle KR. Immunity and inflammation in diabetic kidney disease: translating mechanisms to biomarkers and treatment targets. Am J Physiol Renal Physiol. 2017;312(4):F716–F731. doi:10.1152/ajprenal.00314.2016

    87. Zhan X, Yan C, Chen Y, et al. Celastrol antagonizes high glucose-evoked podocyte injury, inflammation and insulin resistance by restoring the HO-1-mediated autophagy pathway. Mol Immunol. 2018;104:61–68. doi:10.1016/j.molimm.2018.10.021

    88. Hanouneh M, Echouffo Tcheugui JB, Jaar BG. Recent advances in diabetic kidney disease. BMC Med. 2021;19(1):180. doi:10.1186/s12916-021-02050-0

    89. Bellucci R, Martin A, Buren M, Nguyen H-N, Bommarito D, Ritz J. JAK1 and JAK2 modulate myeloma cell susceptibility to NK cells through the interferon gamma (IFN-γ) pathway. Blood. 2011;118(21 Suppl):3960. doi:10.1182/blood.V118.21.3960.3960

    90. Yao M, Mao X, Zhang Z, Cui F, Shao S, Mao B. Communication molecules (ncRNAs) mediate tumor-associated macrophage polarization and tumor progression. Front Cell Dev Biol. 2024;12:1289538. doi:10.3389/fcell.2024.1289538

    91. Swaim CD, Canadeo LA, Huibregtse JM. Approaches for investigating the extracellular signaling function of ISG15. Methods Enzymol. 2019;618:211–227. doi:10.1016/bs.mie.2018.12.027

    92. Kiripolsky J, Romano RA, Kasperek EM, Yu G, Kramer JM. Activation of Myd88-dependent TLRs mediates local and systemic inflammation in a mouse model of primary Sjögren’s syndrome. Front Immunol. 2020;10:2963. doi:10.3389/fimmu.2019.02963

    93. Mussbacher M, Derler M, Basílio J, Schmid JA. NF-κB in monocytes and macrophages – an inflammatory master regulator in multitalented immune cells. Front Immunol. 2023;14:1134661. doi:10.3389/fimmu.2023.1134661

    94. Liang S, Wei H, Sun R, Tian Z. IFNalpha regulates NK cell cytotoxicity through STAT1 pathway. Cytokine. 2003;23(6):190–199. doi:10.1016/s1043-4666(03)00226-6

    95. Fortin C, Huang X, Yang Y. Both NK cell-intrinsic and -extrinsic STAT1 signaling are required for NK cell response against vaccinia virus. J Immunol. 2013;191(1):363–368. doi:10.4049/jimmunol.1202714

    96. Miyagi T, Gil MP, Wang X, Louten J, Chu WM, Biron CA. High basal STAT4 balanced by STAT1 induction to control type 1 interferon effects in natural killer cells. J Exp Med. 2007;204(10):2383–2396. doi:10.1084/jem.20070401

    97. Phatarpekar P, Zhu S, Denman CJ, et al. STAT3 activation promotes NK cell proliferation, NKG2D expression, and NK cell antitumor activity. Blood. 2010;116(21 Suppl):105. doi:10.1182/blood.V116.21.105.105

    Continue Reading

  • A shocking new way to make ammonia, no fossil fuels needed

    A shocking new way to make ammonia, no fossil fuels needed

    University of Sydney researchers have harnessed human-made lightning to develop a more efficient method of generating ammonia – one of the world’s most important chemicals. Ammonia is also the main ingredient of fertilizers that account for almost half of all global food production.

    The team have successfully developed a more straightforward method to produce ammonia (NH3) in gas form. Previous efforts by other laboratories produced ammonia in a solution (ammonium, NH4+), which requires more energy and processes to transform it into the final gas product.

    The current method to generate ammonia, the Haber-Bosch process, comes at great climate cost, leaving a huge carbon footprint. It also needs to happen on a large scale and close to sources of cheap natural gas to make it cost-effective.

    The chemical process that fed the world, and the Sydney team looking to revolutionize it

    Naturally occurring ammonia (mostly in the form of bird droppings), was once so high in demand it fueled wars.

    The invention of the Haber-Bosch process in the 19th century made human-made ammonia possible and revolutionized modern agriculture and industry. Currently 90 percent of global ammonia production relies on the Haber-Bosch process.

    “Industry’s appetite for ammonia is only growing. For the past decade, the global scientific community, including our lab, wants to uncover a more sustainable way to produce ammonia that doesn’t rely on fossil fuels.

    “Currently, generating ammonia requires centralized production and long-distance transportation of the product. We need a low-cost, decentralized and scalable ‘green ammonia’,” said lead researcher Professor PJ Cullen from the University of Sydney’s School of Chemical and Biomolecular Engineering and the Net Zero Institute. His team has been working on ‘green ammonia’ production for six years.

    “In this research we’ve successfully developed a method that allows air to be converted to ammonia in its gaseous form using electricity. A huge step towards our goals.”

    The research was published in AngewandteChemie International edition.

    Ammonia contains three hydrogen molecules, meaning it can be used as an effective carrier and source of hydrogen as an energy source, even potentially as an effective means of storing and transporting hydrogen. Industry bodies have found they can access the hydrogen by ‘cracking’ ammonia to separate the molecules to use the hydrogen.

    Ammonia is also a strong candidate for use as a carbon-free fuel due to its chemical make-up. This has caught the interest of the shipping industry which is responsible for about 3 percent of all global greenhouse gas emissions.

    Cracking a chemical conundrum

    Professor Cullen’s team’s new method to generate ammonia works by harnessing the power of plasma, by electrifying or exciting the air.

    But the star is a membrane-based electrolyser, a seemingly non-descript silver box, where the conversion to gaseous ammonia happens.

    During the Haber-Bosch process, ammonia (NH3) is made by combining nitrogen (N2) and hydrogen (H2) gases under high temperatures and pressure in the presence of catalyst (a substance that speeds up a chemical reaction).

    The plasma-based method Professor Cullen’s team developed uses electricity to excite nitrogen and oxygen molecules in the air. The team then passes these excited molecules to the membrane-based electrolyser to convert the excited molecules to ammonia.

    The researchers said this is a more straightforward pathway for ammonia production.

    Professor Cullen said the findings signal a new phase in making green ammonia possible. The team is now working on making the method more energy efficient and competitive compared to the Haber-Bosch process.

    “This new approach is a two-step process, namely combining plasma and electrolysis. We have already made the plasma component viable in terms of energy efficiency and scalability.

    “To create a more complete solution to a sustainable ammonia productive, we need to push the energy efficiency of the electrolyzer component,” Professor Cullen said.

    Continue Reading

  • Princess Kate’s chat with cancer patients shows precisely why NHS needs change | Royal | News

    Princess Kate’s chat with cancer patients shows precisely why NHS needs change | Royal | News

    On the face of it Princess Kate and I are very different people living very different lives. When I travel home from a gruelling session of chemotherapy there’s always a risk I’ll get stabbed on the tram. If she’s on a tram, it would be for a royal engagement. My cheesy pasta is less about fresh ingredients and more about a jar of pesto and a bag of pre-grated cheese. I’m sure her version is much more wholesome. But her comments on Wednesday, July 2, made me realise that as two people affected by cancer, we are both very similar.

    Everyone’s cancer journey is different, but by speaking out about the mental impact of treatment, and what happens afterwards, Princess Kate said what I try to say, but in a much better way. Speaking to patients at a cancer wellbeing centre at Colchester Hospital, she discussed the emotional impact of fighting the disease.

    Describing it as a “rollercoaster”, Princess Kate said: “There is a whole phase when you finish your treatment, everybody expects you to be better – go! But that’s not the case at all.

    “You’re not necessarily under the clinical team any longer but you’re not able to function normally at home as you perhaps once used to.

    “And actually someone to help talk you through that, show you and guide you through that sort of phase that comes after treatment I think is really valuable.”

    I doubt I will ever get to an “after treatment” phase but my experience last week of being told I don’t have spinal cancer feels pretty close.

    First, there was the worry of thinking I could be dead by Christmas, and then, after scan results came back, the wondering what to do now.

    How do I readjust my life to get back to fighting my incurable bowel cancer while recognising the spinal cancer scare?

    It is the kind of question that thousands of cancer patients grapple with every day. Just what is the best way to tackle the emotional impact of cancer after being left to process good or bad news?

    As Princess Kate says, there isn’t an easy answer. For me, the solution is why I’m leading the Daily Express’s Cancer Care campaign.

    Cancer is the worst thing that most people will experience in their lifetime, and the emotional impact of it can tear people apart, even more than the physical side effects. This is why we are calling for all patients to have mental health support both during and after their treatment.

    Princess Kate recognises just how important this is, and I hope the NHS and the Department of Health listen to her and us and make sure it’s available for all patients across the country.

    Continue Reading

  • Jurassic World Rebirth to Gaza: Doctors Under Attack – the week in rave reviews | Culture

    Jurassic World Rebirth to Gaza: Doctors Under Attack – the week in rave reviews | Culture

    TV

    If you only watch one, make it …

    Gaza: Doctors Under Attack

    Channel 4; available now

    Gaza: Doctors Under Attack. Photograph: Channel 4/Basement Films

    Summed up in a sentence A belated airing of the hugely controversial documentary that the BBC refused to show: a horrifying investigation into claims that Israel’s Defence Force has systematically targeted Palestinian medics.

    What our reviewer said “This is the sort of television that will never leave you. It will provoke an international reaction, and for extremely good cause. Forget what got it stopped at the BBC. It is here now and, regardless of how that happened, we owe it to the subjects to not look away.” Stuart Heritage

    Read the full review

    Further reading Gaza film’s producer accuses BBC of trying to gag him over decision to drop it


    Pick of the rest

    Such Brave Girls

    BBC iPlayer; available now

    Kat Sadler as Josie, Louise Brealey as Deb and Lizzie Davidson as Billie in Such Brave Girls. Photograph: James Stack/BBC/Various Artists

    Summed up in a sentence The second series of a brilliant, startlingly feral comedy about a trio of troubled female relatives – whose first outing won a comedy Bafta.

    What our reviewer said “Such Brave Girls won’t be to everyone’s tastes. But if you like your comedy scary, lairy and perfectly portioned, it is a total knockout.” Hannah J Davies

    Read the full review

    Further reading ‘Who else can we annoy with our show?’: Such Brave Girls, Britain’s most gleefully offensive comedy returns

    Attack on London: Hunting the 7/7 Bombers

    Netflix; full series available

    Summed up in a sentence Interviewees including Tony Blair feature in this absolutely comprehensive look at how the 2005 London transport bombings prompted the UK’s largest criminal investigation.

    What our reviewer said “Though it is by now a familiar story, this evokes the fear, confusion and panic of that day in heart-racing detail.” Rebecca Nicholson

    Read the full review


    You may have missed …

    Shifty

    BBC iPlayer; all episodes available

    Adam Curtis’s Shifty. Photograph: Adam Curtis

    Summed up in a sentence Adam Curtis applies his archive-footage packed documentary style to explaining how the atomisation of UK society has destroyed our democracy – with mesmerising results.

    What our reviewer said “It is an increasing rarity to stand in the presence of anyone with an idea, a thesis, that they have thoroughly worked out to their own satisfaction and then presented stylishly, exuberantly and still intelligently. The hell and the handcart feel that bit more bearable now.” Lucy Mangan

    Read the full review

    Further reading Thatcher, Farage and toe-sucking: Adam Curtis on how Britain came to the brink of civil war


    Film

    If you only watch one, make it …

    Jurassic World Rebirth

    In cinemas now

    Jurassic World Rebirth. Photograph: Universal Pictures and Amblin Entertainment

    Summed up in a sentence Near-extinct franchise roars back to life as latest instalment offers Spielberg-style set pieces and excellent romantic chemistry between leads Scarlett Johansson and Jonathan Bailey.

    What our reviewer said “This new Jurassic adventure isn’t doing anything so very different from the earlier successful models, perhaps, and I could have done without its outrageous brand synergy product placement for certain brands of chocolate bar. But it feels relaxed and sure-footed in its Spielberg pastiche, its big dino-jeopardy moments and its deployment of thrills and laughs.” Peter Bradshaw

    Read the full review

    Further reading ‘The script didn’t have Jurassic World on the front’: Gareth Edwards on Monsters, Godzilla, Star Wars and reinventing dinosaurs


    Pick of the rest

    The Shrouds

    In cinemas now

    Vincent Cassel and Diane Kruger in The Shrouds.

    Summed up in a sentence Elaborate necrophiliac meditation on loss and longing from David Cronenberg, starring Vincent Cassel as an oncologist who has founded a restaurant with a hi-tech cemetery attached.

    What our reviewer said “The film has its own creepy, enveloping mausoleum atmosphere of disquiet, helped by the jarring electronic score by Howard Shore.” Peter Bradshaw

    Read the full review

    Further reading ‘Something must have gone wrong with us’: David Cronenberg and Howard Shore on four decades of body horror

    Hearts of Darkness: A Film-Maker’s Apocalypse

    In cinemas now

    Summed up in a sentence Superb documentary about the making of Francis Ford Coppola’s masterpiece Apocalypse Now, with Coppola’s epic meltdown in the jungle.

    What our reviewer said “Haemorrhaging money and going insanely over-schedule, Coppola shot his film in the Philippines during burning heat, humidity and monsoons and borrowed army helicopters and pilots from President Ferdinand Marcos, only to find that on many occasions – especially during the legendary Ride of the Valkyries attack scene – filming had to halt as the Filipino military would ask for their helicopters back so they could suppress a communist insurgency. In fact, Coppola found himself reproducing reality on a 1:1 scale.” Peter Bradshaw

    Read the full review

    Further reading Francis Ford Coppola: ‘Apocalypse Now is not an anti-war film’


    Now streaming …

    Heads of State

    Prime Video; available now

    Idris Elba and John Cena in Heads of State. Photograph: FlixPix/Alamy

    Summed up in a sentence John Cena and Idris Elba star in fun and well-modulated throwback comedy as the US president and UK prime minister, who team up to escape terrorists.

    What our reviewer said “Fun, fiery and totally frivolous, Heads of State is a perfect summer movie with great potential for future sequels.” Andrew Lawrence

    Read the full review

    Hill

    Sky Cinema and Now; available now

    Damon Hill in Hill. Photograph: Sky

    Summed up in a sentence Compelling story of Formula One star Damon Hill’s trials on and off the racetrack in its depiction of the psychological pressure cooker in which the driver competed.

    What our reviewer said “It has quiet, but profound, lessons to impart in its emphasis on the driver’s need to live up to his roistering father Graham, and on the real meaning of victory in the most alpha of environments that is Formula One.” Phil Hoad

    Read the full review

    Further reading ‘I was angry at the world’: Damon Hill on pain of his father’s death and how it fuelled his rise


    Books

    If you only read one, make it …

    Murderland by Caroline Fraser

    Review by Dorian Lynskey

    Summed up in a sentence An investigation into the causes of America’s 1970s serial killer epidemic comes up with some surprising answers.

    What our reviewer said “It is as hauntingly compulsive a nonfiction book as I have read in a long time. It gets into your blood.”

    Read the full review


    Pick of the rest

    My Sister and Other Lovers by Esther Freud

    Review by Joanna Quinn

    Summed up in a sentence A sequel to Hideous Kinky, 30 years on, explores the effects of an unconventional upbringing.

    What our reviewer said “It’s billed as a novel but arguably occupies an interesting grey area between novel and memoir, resisting the expectations of both and creating something all of its own.”

    Read the full review

    Further reading ‘When I read my sister’s stories I think, that’s not what it was like!’: Esther Freud on the perils of writing about family

    Flashlight by Susan Choi

    Review by Beejay Silcox

    Summed up in a sentence An ambitious, globe-trotting epic of political and family secrets.

    What our reviewer said “Flashlight is all kinds of big: capacious of intent and scope and language and swagger, confronting a chapter of North Korean history that American fiction has barely touched.”

    Read the full review

    Autocorrect by Etgar Keret

    Review by Sam Leith

    Summed up in a sentence Deadpan short stories that range from the surreal to the philosophical to the absurd.

    What our reviewer said “Not so much one book as a library of tiny books, from an author who conveys as well as any I can think of just how much fun you can have with a short story.”

    Read the full review

    Empire of the Elite by Michael M Grynbaum

    Review by Houman Barekat

    Summed up in a sentence Inside the glittering, gossipy world of publisher Condé Nast.

    What our reviewer said “Grynbaum quotes one journalist who believes she missed out on an editorship because, during the interview lunch, she gauchely ate asparagus with cutlery rather than by hand”

    Read the full review


    You may have missed …

    How to Save the Amazon: A journalist’s deadly quest for answers by Dom Phillips

    In bookshops now

    Summed up in a sentence The murdered Guardian journalist’s final investigation, completed by his friends and supporters.

    What our reviewer said “A book both brilliant and broken, one that is ultimately as inspiring and devastating as the Amazon itself” Charlie Gilmour

    Read the full review

    Further reading A deadly mission: how Dom Phillips and Bruno Pereira tried to warn the world about the Amazon’s destruction


    Albums

    If you only listen to one, make it …

    Kesha: . (Period)

    Out now

    Kesha’s album .(Period). Photograph: AP

    Summed up in a sentence After a long legal battle, the pop star’s sixth album harks back to her 2010s hot-mess era, with a buffet of pop styles and only rare hints of her highly publicised trauma.

    What our reviewer said “The songs are all really strong, filled with smart little twists and drops, and funny, self-referential lines.” Alexis Petridis

    Read the full review

    Further reading ‘I would walk in and just cry for two hours’: Kesha on cats, court cases, and the dangers of ‘toxic positivity’


    Pick of the rest

    Daytimers: Alterations

    Out now

    Daytimers’ Alterations LP.

    Summed up in a sentence The UK collective have been reimagining south Asian music since 2020, and their new compilation splices junglism and afro-house on to gems in Sony India’s catalogue.

    What our reviewer said “Reframing this nostalgic cinema music for the modern dancefloor, Alterations proves there is still plenty of space for future generations of diaspora artists to celebrate and find inspiration in their heritage.” Ammar Kalia

    Read the full review

    Kae Tempest: Self Titled

    Out now

    Summed up in a sentence Despair runs through the Londoner’s fifth album but, in what is essentially a love letter to the trans community, his home town and partner, beauty breaks through.

    What our reviewer said “Hope and hard-won happiness, against all odds, underpins this rich, compelling and timely record.” Rachel Aroesti

    Read the full review

    Further reading Kae Tempest: ‘I was living with this boiling hot secret in my heart’

    Shostakovich: Preludes & Fugues Op 87

    Out now

    Summed up in a sentence Performed by Russian pianist Yulianna Avdeeva, these 24 works, modelled on Bach, date from 1950 and 1951 and were originally written for pianist Tatiana Nikolayeva.

    What our reviewer said “Avdeeva takes a lighter approach, less forthright, and perhaps not digging as deeply into the barely disguised tragedy of the E minor Prelude as Nikolayeva does, but equally dazzling in the exuberant display of the A minor.” Andrew Clements

    Read the full review


    On tour this week

    Slayer

    Playing outdoor shows this week

    Slayer perform at Cardiff Castle earlier this week. Photograph: Maxine Howells/Getty Images

    Summed up in a sentence Playing outdoor shows including the big send-off for Black Sabbath on Saturday, the thrash legends have reformed and are playing their first UK gigs in six years.

    What our reviewer said “Slayer are still a shocking proposition, their churning riffs punctuated by gross-out gore and grim images from endless war. Subtle? No. Effective? Absolutely.” Huw Baines

    Read the full review

    Continue Reading

  • Travels with the Queen recalled as royal train nears end of line

    Travels with the Queen recalled as royal train nears end of line

    Danny Fullbrook

    BBC News, Buckinghamshire

    Getty Images Queen Elizabeth II and her husband Prince Philip, Duke of Edinburgh, smile and wave as the Royal train pulls out of Euston n 1977Getty Images

    Queen Elizabeth II and the Duke of Edinburgh smile and wave as the royal train pulls out of Euston in 1977

    Euston Station, 1937: The royal train sits quietly at the platform with a policeman posted outside to guard young Princess Elizabeth; suddenly he hears a soft knock on the carriage window behind him.

    The future Queen beckons him inside. “Here’s a shilling,” she says. “Can you go and get me a comic please?”

    This is one of many anecdotes told by those who worked on the train that are now preserved by journalist and author Phil Marsh.

    He says with a laugh: “Can you imagine being the policeman who’s supposed to be guarding the heir to the throne and then being told to go and buy a comic?”

    Reportedly, the officer did just that.

    royaltrain.co.uk Five carriages of the royal train are visible on a track, behind it is the old red-brick buildings of Wolverton Worksroyaltrain.co.uk

    The train is kept and maintained at Wolverton Works in Buckinghamshire

    In 2027, 90 years after this moment took place, the royal train will be pulled from service.

    Buckingham Palace has taken the decision to decommission the historic rolling stock as part of a “drive to ensure we deliver value for money”.

    It will be taken around the UK before it is removed from service.

    Mr Marsh first became associated with the train in 1997 when he was tasked with putting together a business case to sell it, but he says it “fortunately didn’t stack up”.

    He made friends with Leo Coleman, project manager at Wolverton Works, Buckinghamshire, where the train is kept, who was responsible for modernising the train for the Queen’s silver jubilee in 1977.

    After Mr Coleman died he was left his archive and tasked with chronicling its story, and he has shared some of those memories for this article.

    royaltrain.co.uk Two men smarty dressed are looking at a photograph togetherroyaltrain.co.uk

    Writer Phil Marsh has documented memories from Leo Coleman (left) and Chris Hillyard (right)

    The comic book story was documented by Chris Hillyard, the last foreman of the train, who died in November with cancer.

    On another occasion Mr Hillyard was on the train, alongside the Queen, when he noticed a smell of smoke.

    He stopped the train and asked the signalman to block the adjacent railway while he investigated the fault.

    While he was doing this, the Queen appeared at the window, apparently unaware the other line had been closed.

    She said: “Oh, Mr Hillyard, I’ll be your lookout. It’ll be quite safe.”

    “Yes, ma’am. Thank you,” he responded politely.

    royaltrain.co.uk A black and white photo shows a steam locomotive pulling along the carts of the royal train.royaltrain.co.uk

    Queen Victoria used the royal train during her diamond jubilee in 1897

    Queen Adelaide was the first member of the Royal Family to have a carriage built for the royal train in 1842.

    It continued to be used by members of the Royal Family, including Queen Victoria, who would often stop at Wolverton for a refreshment break as the train did not have toilets.

    In 1869 Wolverton Works built the very first bespoke royal carriages for Queen Victoria, costing £1,800. The monarch donated £800.

    A special shed was constructed for the train in 1869 at Wolverton but has since been converted to flats, though the train has remained at the site for its entire history.

    King Edward VII innovated in 1901 when he introduced electricity, powered by the steam engine, but a generator was eventually installed in 1941 alongside radio and telephones.

    In 1977, when Mr Coleman was tasked with upgrading the royal train for the jubilee, the focus shifted from luxury to function.

    Members of the Royal Family were expected to live and work on the train for long periods, requiring functional design changes such as an office.

    royaltrain.co.uk A photo of the interior of Queen Victoria's carriage on the royal train. There is a long sofa on the left. There are several lamps, two armchairs and decorative curtains on the windows.royaltrain.co.uk

    Phil Marsh described the royal train during the era of Queen Victoria as a ‘palace on wheels’

    The first journey after the 1977 upgrade was from Euston to Glasgow.

    After completing their engagement Queen Elizabeth II and the Duke of Edinburgh asked to speak to Mr Coleman.

    “Everything all right, ma’am?” he asked.

    “No,” she responded, “what’s happened to the old ironing board?”

    As part of the improvements a new ironing board had been installed but the Queen’s lady-in-waiting wanted the old one back.

    Mr Coleman called Wolverton Works and a member of staff had to find it and carry it to Glasgow on the next available train.

    Reuters King Charles III is exiting the train and about to shake hands with a uniformed lady on the platformReuters

    King Charles III has used the royal train a number of times during his reign

    Today, the seven carriages that make up the royal train are owned by Network Rail while the locomotives named King’s Messenger and Royal Sovereign are owned by DB Cargo UK.

    Gemini Rail Services run Wolverton Works, where the train is maintained.

    Engineers from DB Cargo and personnel from Gemini are on board during all journeys in case something goes wrong.

    The seven carriages include a saloon for King Charles, which includes his own bedroom and lounge.

    There is also his day coach, a restaurant cart and a dining cart, and the remaining carts are for use of support staff.

    DB Cargo told the BBC when the royal contract expires on 31 March 2027 it will retain its locomotives and may put them on other traffic.

    Network Rail has been asked what it plans to do with its carriages, but has not yet responded.

    Mr Marsh, who documents the train’s history on the website royaltrain.co.uk, hopes they will be kept in a museum.

    “Every carriage on the train has been designated as part of the national collection,” he explained.

    “Designation means that it can’t be scrapped. It will need to go to a museum whether it’s at York or any other museum is up for debate.”

    Continue Reading

  • Owning dog or cat could preserve some brain functions as we age, study says | Ageing

    Owning dog or cat could preserve some brain functions as we age, study says | Ageing

    As Britain’s population ages and dementia rates climb, scientists may have found an unexpected ally in the fight against cognitive decline.

    Cats and dogs may be exercising more than just your patience: they could be keeping parts of your brain ticking over too. In a potential breakthrough for preventive health, researchers have found that owning a four-pawed friend is linked to slower cognitive decline by potentially preserving specific brain functions as we grow older.

    Interestingly, the associations differ depending on the animal: dog owners were found to retain sharper memory, both immediate and delayed, while cat owners showed slower decline in verbal fluency.

    When it comes to slower cognitive decline in their owners, however, it seems that not all pets are created equal: fish and birds, while charming companions, showed no significant link.

    “Pet ownership has been linked to a positive influence on cognitive functioning and cognitive decline in late adulthood,” said Adriana Rostekova, a researcher and lead author of the article, which was published in Nature. “However, there is limited understanding of how different species of pets are associated with these outcomes.”

    Rostekova, who works at the lifespan developmental psychology research group at the University of Geneva, used data from eight waves of the Survey of Health and Retirement in Europe to examine the relationship between pet ownership and cognitive decline over an 18-year period among adults aged 50 and older.

    She specifically looked at the distinct role of owning dogs, cats, birds and fish. “The key novelty of our study was that we found notable differences between the species,” she said.

    Rostekova hypothesised that because keeping fish or birds showed no meaningful link to changes in cognitive decline, the overall pattern of pet ownership may be driven primarily by having a cat or dog rather than pet ownership in general.

    “Several explanations may help explain the absence of this association in fish and bird owners, despite the reports of their ownership’ positive influence on wellbeing in ways that are usually associated with cognitive benefits,” she added.

    “A fish or bird’s short lifespan may potentially limit the level of emotional connection one is able to develop with the pet fish,” she said. “Bird ownership may negatively affect the owner’s sleep quality due to the increased noise levels, which has been shown to be associated with cognitive decline.”

    Rostekova added: “[It is] further possible that interaction with dogs and cats provides unique cognitive stimulation, which may be less pronounced in other, less demanding pets.”

    Other research has found evidence of an increase in prefrontal brain activation and stronger attentional processes and emotional arousal caused by interaction with a dog.

    There is further evidence of increased activation of the prefrontal cortex and the inferior frontal gyrus when interacting with cats, which is speculated to be linked to the characteristic, hard-to-predict temperament of the animal.

    “There is also a possibility of increased social stimulation facilitated by cats and dogs, which may be linked to the slower cognitive decline experienced by their owners: an increased frequency of social interactions when accompanied by a dog – or for cats, a substitute for a social network,” said Rostekova.

    As the NHS grapples with an ageing population and rising dementia rates, experts say the findings could reshape how we think about healthy ageing – and the animals we choose to age alongside.

    Andrew Scott, the author of The Longevity Imperative and a cat owner (although also a dog lover), said: “We tend to think of health as being about disease and hospitals but as we live longer and need to focus on preventive measures that keep us healthy for longer, we will discover that the health system extends well beyond doctors and hospitals.

    “It is about how we live our life. What is nice about this study is it suggests a fun and meaningful way of keeping healthy and engaged. A lot of things we are recommended to do for our health aren’t always fun or companionable (does anyone fast as a family?). Having a pet can be fun and if it keeps you healthy that’s a great bonus.”

    Continue Reading

  • ‘The vehicle suddenly accelerated with our baby in it’: the terrifying truth about why Tesla’s cars keep crashing | Tesla

    ‘The vehicle suddenly accelerated with our baby in it’: the terrifying truth about why Tesla’s cars keep crashing | Tesla

    It was a Monday afternoon in June 2023 when Rita Meier, 45, joined us for a video call. Meier told us about the last time she said goodbye to her husband, Stefan, five years earlier. He had been leaving their home near Lake Constance, Germany, heading for a trade fair in Milan.

    Meier recalled how he hesitated between taking his Tesla Model S or her BMW. He had never driven the Tesla that far before. He checked the route for charging stations along the way and ultimately decided to try it. Rita had a bad feeling. She stayed home with their three children, the youngest less than a year old.

    At 3.18pm on 10 May 2018, Stefan Meier lost control of his Model S on the A2 highway near the Monte Ceneri tunnel. Travelling at about 100kmh (62mph), he ploughed through several warning markers and traffic signs before crashing into a slanted guardrail. “The collision with the guardrail launches the vehicle into the air, where it flips several times before landing,” investigators would write later.

    The car came to rest more than 70 metres away, on the opposite side of the road, leaving a trail of wreckage. According to witnesses, the Model S burst into flames while still airborne. Several passersby tried to open the doors and rescue the driver, but they couldn’t unlock the car. When they heard explosions and saw flames through the windows, they retreated. Even the firefighters, who arrived 20 minutes later, could do nothing but watch the Tesla burn.

    At that moment, Rita Meier was unaware of the crash. She tried calling her husband, but he didn’t pick up. When he still hadn’t returned her call hours later – highly unusual for this devoted father – she attempted to track his car using Tesla’s app. It no longer worked. By the time police officers rang her doorbell late that night, Meier was already bracing for the worst.

    The crash made headlines the next morning as one of the first fatal Tesla accidents in Europe. Tesla released a statement to the press saying the company was “deeply saddened” by the incident, adding, “We are working to gather all the facts in this case and are fully cooperating with local authorities.”

    To this day, Meier still doesn’t know why her husband died. She has kept everything the police gave her after their inconclusive investigation. The charred wreck of the Model S sits in a garage Meier rents specifically for that purpose. The scorched phone – which she had forensically analysed at her own expense, to no avail – sits in a drawer at home. Maybe someday all this will be needed again, she says. She hasn’t given up hope of uncovering the truth.


    Rita Meier was one of many people who reached out to us after we began reporting on the Tesla Files – a cache of 23,000 leaked documents and 100 gigabytes of confidential data shared by an anonymous whistleblower. The first report we published looked at problems with Tesla’s autopilot system, which allows the cars to temporarily drive on their own, taking over steering, braking and acceleration. Though touted by the company as “Full Self-Driving” (FSD), it is designed to assist, not replace, the driver, who should keep their eyes on the road and be ready to intervene at any time.

    Autonomous driving is the core promise around which Elon Musk has built his company. Tesla has never delivered a truly self-driving vehicle, yet the richest person in the world keeps repeating the claim that his cars will soon drive entirely without human help. Is Tesla’s autopilot really as advanced as he says?

    The Tesla Files suggest otherwise. They contain more than 2,400 customer complaints about unintended acceleration and more than 1,500 braking issues – 139 involving emergency braking without cause, and 383 phantom braking events triggered by false collision warnings. More than 1,000 crashes are documented. A separate spreadsheet on driver-assistance incidents where customers raised safety concerns lists more than 3,000 entries. The oldest date from 2015, the most recent from March 2022. In that time, Tesla delivered roughly 2.6m vehicles with autopilot software. Most incidents occurred in the US, but there have also been complaints from Europe and Asia. Customers described their cars suddenly accelerating or braking hard. Some escaped with a scare; others ended up in ditches, crashing into walls or colliding with oncoming vehicles. “After dropping my son off in his school parking lot, as I go to make a right-hand exit it lurches forward suddenly,” one complaint read. Another said, “My autopilot failed/malfunctioned this morning (car didn’t brake) and I almost rear-ended somebody at 65mph.” A third reported, “Today, while my wife was driving with our baby in the car, it suddenly accelerated out of nowhere.”

    Braking for no reason caused just as much distress. “Our car just stopped on the highway. That was terrifying,” a Tesla driver wrote. Another complained, “Frequent phantom braking on two-lane highways. Makes the autopilot almost unusable.” Some report their car “jumped lanes unexpectedly”, causing them to hit a concrete barrier, or veered into oncoming traffic.

    Musk has given the world many reasons to criticise him since he teamed up with Donald Trump. Many people do – mostly by boycotting his products. But while it is one thing to disagree with the political views of a business leader, it is another to be mortally afraid of his products. In the Tesla Files, we found thousands of examples of why such fear may be justified.

    ‘My husband died in an unexplained accident. And no one cared.’ Illustration: Carl Godfrey/The Guardian

    We set out to match some of these incidents of autopilot errors with customers’ names. Like hundreds of other Tesla customers, Rita Meier entered the vehicle identification number of her husband’s Model S into the response form we published on the website of the German business newspaper Handelsblatt, for which we carried out our investigation. She quickly discovered that the Tesla Files contained data related to the car. In her first email to us, she wrote, “You can probably imagine what it felt like to read that.”

    There isn’t much information – just an Excel spreadsheet titled “Incident Review”. A Tesla employee noted that the mileage counter on Stefan Meier’s car stood at 4,765 miles at the time of the crash. The entry was catalogued just one day after the fatal accident. In the comment field was written, “Vehicle involved in an accident.” The cause of the crash remains unknown to this day. In Tesla’s internal system, a company employee had marked the case as “resolved”, but for five years, Rita Meier had been searching for answers. After Stefan’s death, she took over the family business – a timber company with 200 employees based in Tettnang, Baden-Württemberg. As journalists, we are used to tough interviews, but this one was different. We had to strike a careful balance – between empathy and the persistent questioning good reporting demands. “Why are you convinced the Tesla was responsible for your husband’s death?” we asked her. “Isn’t it possible he was distracted – maybe looking at his phone?”

    No one knows for sure. But Meier was well aware that Musk has previously claimed Tesla “releases critical crash data affecting public safety immediately and always will”; that he has bragged many times about how its superior handling of data sets the company apart from its competitors. In the case of her husband, why was she expected to believe there was no data?

    Meier’s account was structured and precise. Only once did the toll become visible – when she described how her husband’s body burned in full view of the firefighters. Her eyes filled with tears and her voice cracked. She apologised, turning away. After she collected herself, she told us she has nothing left to gain – but also nothing to lose. That was why she had reached out to us. We promised to look into the case.


    Rita Meier wasn’t the only widow to approach us. Disappointed customers, current and former employees, analysts and lawyers were sharing links to our reporting. Many of them contacted us. More than once, someone wrote that it was about time someone stood up to Tesla – and to Elon Musk.

    Meier, too, shared our articles and the callout form with others in her network – including people who, like her, lost loved ones in Tesla crashes. One of them was Anke Schuster. Like Meier, she had lost her husband in a Tesla crash that defies explanation and had spent years chasing answers. And, like Meier, she had found her husband’s Model X listed in the Tesla Files. Once again, the incident was marked as resolved – with no indication of what that actually meant.

    “My husband died in an unexplained and inexplicable accident,” Schuster wrote in her first email. Her dealings with police, prosecutors and insurance companies, she said, had been “hell”. No one seemed to understand how a Tesla works. “I lost my husband. His four daughters lost their father. And no one ever cared.”

    Her husband, Oliver, was a tech enthusiast, fascinated by Musk. A hotelier by trade, he owned no fewer than four Teslas. He loved the cars. She hated them – especially the autopilot. The way the software seemed to make decisions on its own never sat right with her. Now, she felt as if her instincts had been confirmed in the worst way.

    Oliver Schuster was returning from a business meeting on 13 April 2021 when his black Model X veered off highway B194 between Loitz and Schönbeck in north-east Germany. It was 12.50pm when the car left the road and crashed into a tree. Schuster started to worry when her husband missed a scheduled bank appointment. She tried to track the vehicle but found no way to locate it. Even calling Tesla led nowhere. That evening, the police broke the news: after the crash her husband’s car had burst into flames. He had burned to death – with the fire brigade watching helplessly.

    The crashes that killed Meier’s and Schuster’s husbands were almost three years apart but the parallels were chilling. We examined accident reports, eyewitness accounts, crash-site photos and correspondence with Tesla. In both cases, investigators had requested vehicle data from Tesla, and the company hadn’t provided it. In Meier’s case, Tesla staff claimed no data was available. In Schuster’s, they said there was no relevant data.

    Over the next two years, we spoke with crash victims, grieving families and experts around the world. What we uncovered was an ominous black box – a system designed not only to collect and control every byte of customer data, but to safeguard Musk’s vision of autonomous driving. Critical information was sealed off from public scrutiny.


    Elon Musk is a perfectionist with a tendency towards micromanagement. At Tesla, his whims seem to override every argument – even in matters of life and death. During our reporting, we came across the issue of door handles. On Teslas, they retract into the doors while the cars are being driven. The system depends on battery power. If an airbag deploys, the doors are supposed to unlock automatically and the handles extend – at least, that’s what the Model S manual says.

    The idea for the sleek, futuristic design stems from Musk himself. He insisted on retractable handles, despite repeated warnings from engineers. Since 2018, they have been linked to at least four fatal accidents in Europe and the US, in which five people died.

    In February 2024, we reported on a particularly tragic case: a fatal crash on a country road near Dobbrikow, in Brandenburg, Germany. Two 18-year-olds were killed when the Tesla they were in slammed into a tree and caught fire. First responders couldn’t open the doors because the handles were retracted. The teenagers burned to death in the back seat.

    A court-appointed expert from Dekra, one of Germany’s leading testing authorities, later concluded that, given the retracted handles, the incident “qualifies as a malfunction”. According to the report, “the failure of the rear door handles to extend automatically must be considered a decisive factor” in the deaths. Had the system worked as intended, “it is assumed that rescuers might have been able to extract the two backseat passengers before the fire developed further”. Without what the report calls a “failure of this safety function”, the teens might have survived.

    ‘I feel like I’m in the movies’: malfunctioning robotaxi traps passenger in car – video

    Our investigation made waves. The Kraftfahrt-Bundesamt, Germany’s federal motor transport authority, got involved and announced plans to coordinate with other regulatory bodies to revise international safety standards. Germany’s largest automobile club, ADAC, issued a public recommendation that Tesla drivers should carry emergency window hammers. In a statement, ADAC warned that retractable door handles could seriously hinder rescue efforts. Even trained emergency responders, it said, may struggle to reach trapped passengers. Tesla shows no intention of changing the design.

    That’s Musk. He prefers the sleek look of Teslas without handles, so he accepts the risk to his customers. His thinking, it seems, goes something like this: at some point, the engineers will figure out a technical fix. The same logic applies to his grander vision of autonomous driving: because Musk wants to be first, he lets customers test his unfinished Autopilot system on public roads. It’s a principle borrowed from the software world, where releasing apps in beta has long been standard practice. The more users, the more feedback and, over time – often years – something stable emerges. Revenue and market share arrive much earlier. The motto: if you wait, you lose.

    Musk has taken that mindset to the road. The world is his lab. Everyone else is part of the experiment.


    By the end of 2023, we knew a lot about how Musk’s cars worked – but the way they handle data still felt like a black box. How is that data stored? At what moment does the onboard computer send it to Tesla’s servers? We talked to independent experts at the Technical University Berlin. Three PhD candidates – Christian Werling, Niclas Kühnapfel and Hans Niklas Jacob – made headlines for hacking Tesla’s autopilot hardware. A brief voltage drop on a circuit board turned out to be just enough to trick the system into opening up.

    The security researchers uncovered what they called “Elon Mode” – a hidden setting in which the car drives fully autonomously, without requiring the driver to keep his hands on the wheel. They also managed to recover deleted data, including video footage recorded by a Tesla driver. And they traced exactly what data Tesla sends to its servers – and what it doesn’t.

    The hackers explained that Tesla stores data in three places. First, on a memory card inside the onboard computer – essentially a running log of the vehicle’s digital brain. Second, on the event data recorder – a black box that captures a few seconds before and after a crash. And third, on Tesla’s servers, assuming the vehicle uploads them.

    The researchers told us they had found an internal database embedded in the system – one built around so-called trigger events. If, for example, the airbag deploys or the car hits an obstacle, the system is designed to save a defined set of data to the black box – and transmit it to Tesla’s servers. Unless the vehicles were in a complete network dead zone, in both the Meier and Schuster cases, the cars should have recorded and transmitted that data.

    ‘Is the car driving erratically by itself normal? Yeah, that happens every now and then.’ Illustration: Carl Godfrey/The Guardian

    Who in the company actually works with that data? We examined testimony from Tesla employees in court cases related to fatal crashes. They described how their departments operate. We cross-referenced their statements with entries in the Tesla Files. A pattern took shape: one team screens all crashes at a high level, forwarding them to specialists – some focused on autopilot, others on vehicle dynamics or road grip. There’s also a group that steps in whenever authorities request crash data.

    skip past newsletter promotion

    We compiled a list of employees relevant to our reporting. Some we tried to reach by email or phone. For others, we showed up at their homes. If they weren’t there, we left handwritten notes. No one wanted to talk.

    We searched for other crashes. One involved Hans von Ohain, a 33-year-old Tesla employee from Evergreen, Colorado. On 16 May 2022, he crashed into a tree on his way home from a golf outing and the car burst into flames. Von Ohain died at the scene. His passenger survived and told police that von Ohain, who had been drinking, had activated Full Self-Driving. Tesla, however, said it couldn’t confirm whether the system was engaged – because no vehicle data was transmitted for the incident.

    Then, in February 2024, Musk himself stepped in. The Tesla CEO claimed von Ohain had never downloaded the latest version of the software – so it couldn’t have caused the crash. Friends of von Ohain, however, told US media he had shown them the system. His passenger that day, who barely escaped with his life, told reporters that hours earlier the car had already driven erratically by itself. “The first time it happened, I was like, ‘Is that normal?’” he recalled asking von Ohain. “And he was like, ‘Yeah, that happens every now and then.’”

    His account was bolstered by von Ohain’s widow, who explained to the media how overjoyed her husband had been at working for Tesla. Reportedly, von Ohain received the Full Self-Driving system as a perk. His widow explained how he would use the system almost every time he got behind the wheel: “It was jerky, but we were like, that comes with the territory of new technology. We knew the technology had to learn, and we were willing to be part of that.”

    The Colorado State Patrol investigated but closed the case without blaming Tesla. It reported that no usable data was recovered.


    For a company that markets its cars as computers on wheels, Tesla’s claim that it had no data available in all these cases is surprising. Musk has long described Tesla vehicles as part of a collective neural network – machines that continuously learn from one another. Think of the Borg aliens from the Star Trek franchise. Musk envisions his cars, like the Borg, as a collective – operating as a hive mind, each vehicle linked to a unified consciousness.

    When a journalist asked him in October 2015 what made Tesla’s driver-assistance system different, he replied, “The whole Tesla fleet operates as a network. When one car learns something, they all learn it. That is beyond what other car companies are doing.” Every Tesla driver, he explained, becomes a kind of “expert trainer for how the autopilot should work”.

    According to Musk, the eight cameras in every Tesla transmit more than 160bn video frames a day to the company’s servers. In its owner’s manual, Tesla states that its cars may collect even more: “analytics, road segment, diagnostic and vehicle usage data”, all sent to headquarters to improve product quality and features such as autopilot. The company claims it learns “from the experience of billions of miles that Tesla vehicles have driven”.

    It is a powerful promise: a fleet of millions of cars, constantly feeding raw information into a gargantuan processing centre. Billions – trillions – of data points, all in service of one goal: making cars drive better and keeping drivers safe. At the start of this year, Musk got a chance to show the world what he meant.

    On 1 January 2025, at 8.39am, a Tesla Cybertruck exploded outside the Trump International Hotel Las Vegas. The man behind the incident – US special forces veteran Matthew Livelsberger – had rented the vehicle, packed it with fireworks, gas canisters and grenades, and parked it in front of the building. Just before the explosion, he shot himself in the head with a .50 calibre Desert Eagle pistol. “This was not a terrorist attack, it was a wakeup call. Americans only pay attention to spectacles and violence,” Livelsberger wrote in a letter later found by authorities. “What better way to get my point across than a stunt with fireworks and explosives.”

    The soldier miscalculated. Seven bystanders suffered minor injuries. The Cybertruck was destroyed, but not even the windows of the hotel shattered. Instead, with his final act, Livelsberger revealed something else entirely: just how far the arm of Tesla’s data machinery can reach. “The whole Tesla senior team is investigating this matter right now,” Musk wrote on X just hours after the blast. “Will post more information as soon as we learn anything. We’ve never seen anything like this.”

    Later that day, Musk posted again. Tesla had already analysed all relevant data – and was ready to offer conclusions. “We have now confirmed that the explosion was caused by very large fireworks and/or a bomb carried in the bed of the rented Cybertruck and is unrelated to the vehicle itself,” he wrote. “All vehicle telemetry was positive at the time of the explosion.”

    Suddenly, Musk wasn’t just a CEO; he was an investigator. He instructed Tesla technicians to remotely unlock the scorched vehicle. He handed over internal footage captured up to the moment of detonation.The Tesla CEO had turned a suicide attack into a showcase of his superior technology.

    Yet there were critics even in the moment of glory. “It reveals the kind of sweeping surveillance going on,” warned David Choffnes, executive director of the Cybersecurity and Privacy Institute at Northeastern University in Boston, when contacted by a reporter. “When something bad happens, it’s helpful, but it’s a double-edged sword. Companies that collect this data can abuse it.”

    ‘In many crashes, investigators weren’t even aware that requesting data from Tesla was an option.’ Illustration: Carl Godfrey/The Guardian

    There are other examples of what Tesla’s data collection makes possible. We found the case of David and Sheila Brown, who died in August 2020 when their Model 3 ran a red light at 114mph in Saratoga, California. Investigators managed to reconstruct every detail, thanks to Tesla’s vehicle data. It shows exactly when the Browns opened a door, unfastened a seatbelt, and how hard the driver pressed the accelerator – down to the millisecond, right up to the moment of impact. Over time, we found more cases, more detailed accident reports. The data definitely is there – until it isn’t.

    In many crashes when Teslas inexplicably veered off the road or hit stationary objects, investigators didn’t actually request data from the company. When we asked authorities why, there was often silence. Our impression was that many prosecutors and police officers weren’t even aware that asking was an option. In other cases, they acted only when pushed by victims’ families.

    In the Meier case, Tesla told authorities, in a letter dated 25 June 2018, that the last complete set of vehicle data was transmitted nearly two weeks before the crash. The only data from the day of the accident was a “limited snapshot of vehicle parameters” – taken “approximately 50 minutes before the incident”. However, this snapshot “doesn’t show anything in relation to the incident”. As for the black box, Tesla warned that the storage modules were likely destroyed, given the condition of the burned-out vehicle. Data transmission after a crash is possible, the company said – but in this case, it didn’t happen. In the end, investigators couldn’t even determine whether driver-assist systems were active at the time of the crash.

    The Schuster case played out similarly. Prosecutors in Stralsund, Germany, were baffled. The road where the crash happened is straight, the asphalt was dry and the weather at the time of the accident was clear. Anke Schuster kept urging the authorities to examine Tesla’s telemetry data.

    When prosecutors did formally request the data recorded by Schuster’s car on the day of the crash, it took Tesla more than two weeks to respond – and when it did, the answer was both brief and bold. The company didn’t say there was no data. It said that there was “no relevant data”. The authorities’ reaction left us stunned. We expected prosecutors to push back – to tell Tesla that deciding what’s relevant is their job, not the company’s. But they didn’t. Instead, they closed the case.

    The hackers from TU Berlin pointed us to a study by the Netherlands Forensic Institute, an independent division of the ministry of justice and security. In October 2021, the NFI published findings showing it had successfully accessed the onboard memories of all major Tesla models. The researchers compared their results with accident cases in which police had requested data from Tesla. Their conclusion was that while Tesla formally complied with those requests, it omitted large volumes of data that might have proved useful.

    Tesla’s credibility took a further hit in a report released by the US National Highway Traffic Safety Administration in April 2024. The agency concluded that Tesla failed to adequately monitor whether drivers remain alert and ready to intervene while using its driver-assist systems. It reviewed 956 crashes, field data and customer communications, and pointed to “gaps in Tesla’s telematic data” that made it impossible to determine how often autopilot was active during crashes. If a vehicle’s antenna was damaged or it crashed in an area without network coverage, even serious accidents sometimes went unreported. Tesla’s internal statistics include only those crashes in which an airbag or other pyrotechnic system deployed – something that occurs in just 18% of police-reported cases. This means that the actual accident rate is significantly higher than Tesla discloses to customers and investors.

    There’s more. Two years prior, the NHTSA had flagged something strange – something suspicious. In a separate report, it documented 16 cases in which Tesla vehicles crashed into stationary emergency vehicles. In each, autopilot disengaged “less than one second before impact” – far too little time for the driver to react. Critics warn that this behaviour could allow Tesla to argue in court that autopilot was not active at the moment of impact, potentially dodging responsibility.

    The YouTuber Mark Rober, a former engineer at Nasa, replicated this behaviour in an experiment on 15 March 2025. He simulated a range of hazardous situations, in which the Model Y performed significantly worse than a competing vehicle. The Tesla repeatedly ran over a crash-test dummy without braking. The video went viral, amassing more than 14m views within a few days.

    Mark Rober’s Tesa test drive

    The real surprise came after the experiment. Fred Lambert, who writes for the blog Electrek, pointed out the same autopilot disengagement that the NHTSA had documented. “Autopilot appears to automatically disengage a fraction of a second before the impact as the crash becomes inevitable,” Lambert noted.

    And so the doubts about Tesla’s integrity pile up. In the Tesla Files, we found emails and reports from a UK-based engineer who led Tesla’s Safety Incident Investigation programme, overseeing the company’s most sensitive crash cases. His internal memos reveal that Tesla deliberately limited documentation of particular issues to avoid the risk of this information being requested under subpoena. Although he pushed for clearer protocols and better internal processes, US leadership resisted – explicitly driven by fears of legal exposure.

    We contacted Tesla multiple times with questions about the company’s data practices. We asked about the Meier and Schuster cases – and what it means when fatal crashes are marked “resolved” in Tesla’s internal system. We asked the company to respond to criticism from the US traffic authority and to the findings of Dutch forensic investigators. We also asked why Tesla doesn’t simply publish crash data, as Musk once promised to do, and whether the company considers it appropriate to withhold information from potential US court orders. Tesla has not responded to any of our questions.

    Elon Musk boasts about the vast amount of data his cars generate – data that, he claims, will not only improve Tesla’s entire fleet but also revolutionise road traffic. But, as we have witnessed again and again in the most critical of cases, Tesla refuses to share it.

    Tesla’s handling of crash data affects even those who never wanted anything to do with the company. Every road user trusts the car in front, behind or beside them not to be a threat. Does that trust still stand when the car is driving itself?

    Internally, we called our investigation into Tesla’s crash data Black Box. At first, because it dealt with the physical data units built into the vehicles – so-called black boxes. But the devices Tesla installs hardly deserve the name. Unlike the flight recorders used in aviation, they’re not fireproof – and in many of the cases we examined, they proved useless.

    Over time, we came to see that the name held a second meaning. A black box, in common parlance, is something closed to the outside. Something opaque. Unknowable. And while we’ve gained some insight into Tesla as a company, its handling of crash data remains just that: a black box. Only Tesla knows how Elon Musk’s vehicles truly work. Yet today, more than 5m of them share our roads.

    Some names have been changed.

    This is an edited extract from The Tesla Files by Sönke Iwersen and Michael Verfürden, published on 24 July by Penguin Michael Joseph at £22. To support the Guardian, order your copy at guardianbookshop.com. Delivery charges may apply.

    Continue Reading

  • Photos of the global life of the Dalai Lama as he turns 90

    Photos of the global life of the Dalai Lama as he turns 90

    DHARAMSHALA, India (AP) — The Dalai Lama is revered as a deity by millions of Tibetan Buddhists and known worldwide as a resolute voice for peace, spirituality and Tibet ’s autonomy. He is also seen as a threat by China, which accuses him of wanting to wrest Tibet from Beijing’s control.

    As the spiritual and political leader of Tibetan Buddhists, he established a government-in-exile in the Indian town of Dharamshala after fleeing Tibet in 1959. Since then he has traveled the world to raise the issue of Tibet and Tibetans, while spreading a message of nonviolence.

    He has met world leaders and celebrities, from the likes of fellow Nobel Peace Prize winners Nelson Mandela and Desmond Tutu to multiple U.S. presidents, popes and Hollywood stars.

    Tibetan spiritual leader the Dalai Lama listens to questions during a news conference in London on March 20, 1991. (AP Photo/Dave Caulkin, File)

    Television talk show host Tom Snyder, left, shares a joke with Tibetan spiritual leader the Dalai Lama during the taping of NBC's The Tomorrow Show in New York, Sept. 5, 1979. (AP Photo/Dan Grossi, File)

    Television talk show host Tom Snyder, left, shares a joke with Tibetan spiritual leader the Dalai Lama during the taping of NBC’s The Tomorrow Show in New York, Sept. 5, 1979. (AP Photo/Dan Grossi, File)

    Tibetan spiritual leader the Dalai Lama, center, is flanked by actor and activist Richard Gere, left, and model and actress Cindy Crawford, at a dinner to benefit the American Himalayan Foundation at the Beverly Wilshire Hotel in Beverly Hills, Ca., Sept. 17, 1993. (AP Photo/Reed Saxon, File)

    Tibetan spiritual leader the Dalai Lama, center, is flanked by actor and activist Richard Gere, left, and model and actress Cindy Crawford, at a dinner to benefit the American Himalayan Foundation at the Beverly Wilshire Hotel in Beverly Hills, Ca., Sept. 17, 1993. (AP Photo/Reed Saxon, File)

    Pope John Paul II, left, meets with Tibetan spiritual leader the Dalai Lama at the Vatican Nunciature, embassy in Vatican City on Feb. 2, 1986. (AP Photo/Arturo Mari, File)

    Pope John Paul II, left, meets with Tibetan spiritual leader the Dalai Lama at the Vatican Nunciature, embassy in Vatican City on Feb. 2, 1986. (AP Photo/Arturo Mari, File)

    President George Bush, left, and Tibetan spiritual leader the Dalai Lama look up in the Capitol Rotunda on Capitol Hill in Washington, Oct. 17, 2007, during the Congressional Gold Medal ceremony honoring The Dalai Lama. (AP Photo/Pablo Martinez Monsivais, File)

    President George Bush, left, and Tibetan spiritual leader the Dalai Lama look up in the Capitol Rotunda on Capitol Hill in Washington, Oct. 17, 2007, during the Congressional Gold Medal ceremony honoring The Dalai Lama. (AP Photo/Pablo Martinez Monsivais, File)

    Mother Teresa of Calcutta, right, meets with Tibetan spiritual leader the Dalai Lama, at the Global Survival Conference in Oxford, England, April 12, 1988. (AP Photo/Dave Caulkin, File)

    Mother Teresa of Calcutta, right, meets with Tibetan spiritual leader the Dalai Lama, at the Global Survival Conference in Oxford, England, April 12, 1988. (AP Photo/Dave Caulkin, File)

    Tibetan spiritual leader the Dalai Lama, left, walks hand-in-hand with South African President Nelson Mandela prior to an official reception at the presidential office in Cape Town, August 22, 1996. (AP Photo/Sasa Kralj, File)

    Tibetan spiritual leader the Dalai Lama, left, walks hand-in-hand with South African President Nelson Mandela prior to an official reception at the presidential office in Cape Town, August 22, 1996. (AP Photo/Sasa Kralj, File)

    Tibetan spiritual leader the Dalai Lama, left, displays the Nobel Peace Prize after receiving it from Egil Aarvik, Chairman of the Nobel Committee, at Oslo University's Avla Hall in Norway, Dec. 10, 1989. (AP Photo/Pool, Norwegian News Agency, Inge Gjellesvik, File)

    Tibetan spiritual leader the Dalai Lama, left, displays the Nobel Peace Prize after receiving it from Egil Aarvik, Chairman of the Nobel Committee, at Oslo University’s Avla Hall in Norway, Dec. 10, 1989. (AP Photo/Pool, Norwegian News Agency, Inge Gjellesvik, File)

    House Speaker Nancy Pelosi of Calif. touches heads with Tibetan spiritual leader the Dalai Lama, right, who received the first Lantos Human Rights Prize named for the late California Rep. Tom Lantos on Capitol Hill in Washington, Oct. 6, 2009. (AP Photo/Harry Hamburg, File)

    House Speaker Nancy Pelosi of Calif. touches heads with Tibetan spiritual leader the Dalai Lama, right, who received the first Lantos Human Rights Prize named for the late California Rep. Tom Lantos on Capitol Hill in Washington, Oct. 6, 2009. (AP Photo/Harry Hamburg, File)

    Nobel Peace Prize winner and former Polish President Lech Walesa, left, and fellow laureates Tibetan spiritual leader the Dalai Lama, center, and Adolfo Perez Esquivel greet each other in Gdansk, Poland, Dec. 5, 2008. (AP Photo/Czarek Sokolowski, File)

    Nobel Peace Prize winner and former Polish President Lech Walesa, left, and fellow laureates Tibetan spiritual leader the Dalai Lama, center, and Adolfo Perez Esquivel greet each other in Gdansk, Poland, Dec. 5, 2008. (AP Photo/Czarek Sokolowski, File)

    Tibetan spiritual leader the Dalai Lama is welcomed as he arrives in Graz airport, June 25, 1995. (AP Photo/Gepa, File)

    Tibetan spiritual leader the Dalai Lama is welcomed as he arrives in Graz airport, June 25, 1995. (AP Photo/Gepa, File)

    Tibetan spiritual leader the Dalai Lama, second right poses for a photograph with former President of Afghanistan Sibghatullah Al Mojaddedi, second left, Delhi Jama Masjid high priest Syed Ahmed Bukhari and Sikh leader jathedar Avtar Singh, right, at an anti-terrorism Conference organized by Muslim organizations in New Delhi, India, June 1, 2008. (AP Photo/Manish Swarup, File)

    Tibetan spiritual leader the Dalai Lama, second right poses for a photograph with former President of Afghanistan Sibghatullah Al Mojaddedi, second left, Delhi Jama Masjid high priest Syed Ahmed Bukhari and Sikh leader jathedar Avtar Singh, right, at an anti-terrorism Conference organized by Muslim organizations in New Delhi, India, June 1, 2008. (AP Photo/Manish Swarup, File)

    Tibetan spiritual leader the Dalai Lama greets Lady Gaga, right, before a question and answer session at the U.S. Conference of Mayors in Indianapolis, June 26, 2016. (AP Photo/Michael Conroy, File)

    Tibetan spiritual leader the Dalai Lama greets Lady Gaga, right, before a question and answer session at the U.S. Conference of Mayors in Indianapolis, June 26, 2016. (AP Photo/Michael Conroy, File)

    German Chancellor Angela Merkel, front right, and Tibetan spiritual leader the Dalai Lama leave after a meeting at the chancellery in Berlin on Sept. 23, 2007. (AP Photo/Markus Schreiber, Pool, File)

    German Chancellor Angela Merkel, front right, and Tibetan spiritual leader the Dalai Lama leave after a meeting at the chancellery in Berlin on Sept. 23, 2007. (AP Photo/Markus Schreiber, Pool, File)

    Tibetan spiritual leader the Dalai Lama, left, holds actress Whoopi Goldberg's hand during the World Peace event on the West Lawn of the U.S. Capitol in Washington, July 9, 2011. (AP Photo/Luis M. Alvarez, File)

    Tibetan spiritual leader the Dalai Lama, left, holds actress Whoopi Goldberg’s hand during the World Peace event on the West Lawn of the U.S. Capitol in Washington, July 9, 2011. (AP Photo/Luis M. Alvarez, File)

    President George Bush, front right, and others, pose with Tibetan spiritual leader the Dalai Lama, in yellow robe, during the Congressional Gold Medal Ceremony Honoring the Dalai Lama in the Capitol Rotunda on Capitol Hill in Washington, Oct. 17, 2007. (AP Photo/Pablo Martinez Monsivais, File)

    President George Bush, front right, and others, pose with Tibetan spiritual leader the Dalai Lama, in yellow robe, during the Congressional Gold Medal Ceremony Honoring the Dalai Lama in the Capitol Rotunda on Capitol Hill in Washington, Oct. 17, 2007. (AP Photo/Pablo Martinez Monsivais, File)

    Tibetan spiritual leader the Dalai Lama, left, receives a souvenir from the former Indian prime minister Manmohan Singh in New Delhi, India, Jan. 4, 2016. (AP Photo /Tsering Topgyal, File)

    Tibetan spiritual leader the Dalai Lama, left, receives a souvenir from the former Indian prime minister Manmohan Singh in New Delhi, India, Jan. 4, 2016. (AP Photo /Tsering Topgyal, File)

    Retired Archbishop Desmond Tutu, left and Tibetan spiritual leader the Dalai Lama share a lighter moment as they interact with children at the Tibetan Children's Village School in Dharamshala, India, April 23, 2015. (AP Photo/Ashwini Bhatia, File)

    Retired Archbishop Desmond Tutu, left and Tibetan spiritual leader the Dalai Lama share a lighter moment as they interact with children at the Tibetan Children’s Village School in Dharamshala, India, April 23, 2015. (AP Photo/Ashwini Bhatia, File)

    Tibetan spiritual leader the Dalai Lama speaks during a conference on Quantum Physics and Madhyamika Philosophical View, in New Delhi, India, Nov. 12, 2015. (AP Photo /Tsering Topgyal, File)

    Tibetan spiritual leader the Dalai Lama speaks during a conference on Quantum Physics and Madhyamika Philosophical View, in New Delhi, India, Nov. 12, 2015. (AP Photo /Tsering Topgyal, File)

    Tibetan spiritual leader the Dalai Lama, right, greets a Buddhist devotee as he arrives at the Tibetan Children's Village school near Leh, India, July 3, 2014. (AP Photo/Tsering Topgyal, File)

    Tibetan spiritual leader the Dalai Lama, right, greets a Buddhist devotee as he arrives at the Tibetan Children’s Village school near Leh, India, July 3, 2014. (AP Photo/Tsering Topgyal, File)

    Tibetan spiritual leader the Dalai Lama slings a t-shirt signed by the Australian cricket team players as he poses for a photograph with the team at the Tsuglakhang temple in Dharamshala, India, March 24, 2017. (AP Photo/Ashwini Bhatia, File)

    Tibetan spiritual leader the Dalai Lama slings a t-shirt signed by the Australian cricket team players as he poses for a photograph with the team at the Tsuglakhang temple in Dharamshala, India, March 24, 2017. (AP Photo/Ashwini Bhatia, File)

    As he celebrates his 90th birthday on Sunday, The Associated Press has curated a selection of photos of the Dalai Lama, from his early days in India to appearances he has made around the world.

    ___

    This is a photo gallery curated by AP photo editors.


    Continue Reading

  • Transcendent Spaces: The Role of Museums in Medical Education on Relig

    Transcendent Spaces: The Role of Museums in Medical Education on Relig

    Correspondence: Eojin Choi, Johns Hopkins University School of Medicine, 733 N Broadway, Baltimore, MD, 21205, USA, Email [email protected]

    Abstract: Religion and spirituality are increasingly recognized as important aspects of patient care and medical education, yet many medical schools still lack structured curricula in this area. This is particularly relevant given the increasing gap between younger medical learners who identify as “spiritual but not religious” and their older adult patients who identify as religious. This article explores the potential of museum-based education as an innovative approach to integrate religion and spirituality into medical education. By using museums’ diverse collections of religious and cultural artifacts, medical students can learn about various religious traditions around the world and engage in discussions on religion and spirituality in a collaborative and supportive environment. Visual Thinking Strategies, a widely studied visual arts-based method in medical education, can be a particularly effective tool that fosters empathy, cultural humility, and critical thinking. This approach can ultimately help medical students integrate spiritual care into their future practice while also encouraging reflection on the role of religion and spirituality in their personal lives.

    Keywords: medical education, museum-based education, visual thinking strategies, religion, spirituality, spiritual care

    Introduction

    Throughout human history, individuals have sought meaning in something greater than themselves, a search that intensifies for patients and families during healthcare crises. With 71% of young adults (ages 18–29 years) in the United States identifying as either religious or “spiritual but not religious”, young adult medical students are likely to have an interest in reflecting on religion and spirituality in their personal lives and/or a desire to learn how to provide spiritual care for their patients.1 Spirituality has been defined as “the personal quest for understanding answers to ultimate questions about life, about meaning, and about relationship to the sacred or transcendent, which may (or may not) lead to or arise from the development of religious rituals and the formation of community”.2 Religion, on the other hand, involves a search for the sacred and non-sacred goals (such as identity and meaning) within a structured community, often with institutional beliefs, practices, and rituals.3 Spiritual care is the process of identifying and addressing the spiritual needs of patients, whether they are secular, spiritual, and/or religious.4

    Despite the importance of religion and spirituality to both personal growth and the practice of medicine, medical students have surprisingly few opportunities to engage in these topics as part of their formal education. Thus, little is known about how best to explore these human experiences—and the enduring questions they raise—with medical students. In this article, we explore the potential of museum-based education as an innovative way to integrate religion and spirituality into medical education.

    The Evidence About Religion and Spirituality in Medical Education

    While medical schools have increasingly incorporated spirituality and health into their curricula, these programs vary widely and have not been well described and/or evaluated, with some notable exceptions.5–7 A systematic review of the literature from 1926 to 2020 that included 19 publications found that only around half the courses were mandatory (11/20) and had a pre- and post-test design (11/20), while only three studies assessed the long-term outcomes of the course.5 Many courses included education on taking a spiritual history and the role of chaplains in spiritual care, often incorporating chaplain shadowing opportunities as well as reflective writing.5

    Another systematic review on spiritual care training programs for students or healthcare professionals identified several barriers to integrating spiritual care into healthcare, such as negative perceptions on spirituality, spiritual care not being viewed as a priority, and a resistance to examining one’s own spirituality.7 A scoping review of religion and spirituality in residents (and inter-relationships with clinical practice and residency training) found that only about 40% of residents reported receiving education on religion and spirituality during medical school and, not surprisingly, many felt they lacked both the knowledge and skills to address these topics with their patients.8 Thus, more research is needed to guide curricular development and evaluate long-term outcomes.

    Although most young adults identify as either religious or “spiritual but not religious”, these numbers are not static, and the gap is growing between the percentage of young adults who identify as religious and older adults who identify as religious.9 In 2007, 74% of young adults (ie, under 30 years of age) and 92% of older adults (ages 65 years or older) identified with a religion.9 In 2023–24, 54% of young adults and 83% of older adults identified with a religion.9 This trend suggests a significant and widening gap in religious identification between young adults and older adults. In turn, this may reflect a similar gulf in religious beliefs between medical students and the older patients they will serve.

    For many older patients, religion is an important part of their lives, especially during their sickest and most vulnerable moments. In 2023–24, it was reported that 49% of older adults consider religion to be “very important” in their lives, and 55% pray at least once daily.9 This suggests that religion and spirituality are important aspects of being human for many patients and are therefore relevant in some way to all students who care about the well-being of their patients. The nature of health and healing, the role of suffering, what it means to live well and die well are all important, enduring questions whose answers often depend on one’s religious and spiritual beliefs. For example, healthcare providers may need to navigate challenging situations involving patients or surrogate decision makers who refuse blood transfusions or make decisions regarding pregnancy termination based on their religious beliefs. Thus, it is crucial for medical students to explore these questions and develop cultural competence and safety (an important aspect of the core clinical competencies) as they learn to work with patients of various identities and backgrounds.10,11 Cultural safety is a patient-centered approach that emphasizes the need for providers to reflect on cultural identities as well as their own cultural biases, understand the impact of power imbalances, and create an environment where patients feel respected and empowered.11,12 A lack of cultural competence and safety regarding religion and spirituality can negatively impact patient-physician relationships and patient-centered approaches to treatment.13 But how best to educate students on religion and spirituality in a way that is psychologically safe, engaging, and open to diversity?

    Museum-Based Education on Religion and Spirituality

    One innovative approach to providing medical education on spirituality and cultural humility involves museums. Museums are full of “third things”, which can be defined as objects, artwork, texts, and other types of media that provide a mediating focal point for reflection and conversation, thus helping create a safe space for openly discussing different perspectives.14 “Third things” can be especially helpful when facilitating discussions about difficult topics by balancing vulnerability and emotional safety, as participants can choose to share personal stories or to focus more on the “third thing” if the topic is too personal or painful.15,16 In medical education, “third things” have been used to help foster empathy, provide opportunities to reflect, and renew a sense of meaning among learners.17

    As a substantial proportion of the art in museums is religious in nature, these collections can provide opportunities to explore various religious traditions around the world.18 Art and religion have often been intertwined throughout history, beginning with the use of religious objects and art to conduct rituals and decorate sacred places.19 For example, rituals and religious beliefs—such as the belief in the afterlife—significantly contributed to the development of Egyptian art.20 In recent decades, museum exhibitions have addressed the major religions of the world and showcased works from specific faiths in their cultural and historical contexts.19

    Thus, museums can serve as transcendent spaces that cultivate introspection, especially on topics related to religion and spirituality. Immersion in museum exhibits offers opportunities for “aesthetic awareness”, where engaging deeply with art fosters connection and self-actualization, and “numinous experiences”, which are moments of transcendence that can inspire emotions such as grief, joy, or wonder.21,22 Museums have increasingly embraced this role and have shifted, as museum scholar Stephen Weil stated, “From being about something to being for somebody”.23 In particular, Visual Thinking Strategies (VTS), a well-studied visual arts-based teaching method, can encourage students to reflect and share their insights in a dynamic and collaborative environment. In a VTS session, participants first observe a work of art in silence and then engage in a group discussion guided by three specific questions: (1) What’s going on in this picture? (2) What do you see that makes you say that? and (3) What more can we find? These questions are designed to encourage participants to observe closely, ground their interpretations in visual evidence, and persistently engage in open-ended inquiry.24 VTS has been shown to help promote crucial skills and characteristics important for clinical practice, including empathy, observation, communication skills, cultural sensitivity, and tolerance for ambiguity.25 One study revealed that an art museum-based program helped clinical-level medical students gain a deeper awareness and progression of their professional identity.26

    Spiritual care is relevant not only for patients’ health and quality of life but also for patients’ relatives, partners, and friends who may be caregivers and/or experiencing grief. As chaplains are integral members of the interdisciplinary healthcare team, clinicians and medical students can work with them to contribute to spiritual care. Moreover, museum-based education allows medical students to pause and reflect on religion and spirituality in their personal lives and clinical settings. As spirituality can be described as universal yet deeply personal in nature, these opportunities would allow students to reflect on their own understandings and perspectives on the meaning of spirituality. This reflective practice can potentially act as a protective factor against burnout, especially after emotionally challenging patient encounters.27

    Discussion

    Our article explores museum-based education as an innovative and impactful approach to integrating religion and spirituality into medical education. Using selected artwork and artifacts as “third things”, educators can design activities that foster deep reflection and discussions. This approach supports development of both technical and non-technical skills such as observation, communication, and empathy while also providing opportunities for personal insights. Additionally, group discussions in museum settings are often supportive and enhance appreciation of multiple perspectives.

    However, this approach also has several limitations. First, it requires training in facilitation to ensure that discussions remain inclusive and meaningful.28 Second, evidence on the long-term impact of museum-based programs is limited.5 Third, more research in museum-based education for medical learners, especially regarding religion and spirituality, is needed. We also recognize that not everyone has access to museums in their community—however, many museum-based learning activities, like VTS, have been adapted successfully to classroom and virtual settings.29–31 Both in-person and virtual formats offer unique benefits, as virtual options offer increased accessibility and comfort for some learners while others may find in-person experiences to be more engaging and powerful.31

    The advent and adoption of virtual reality (VR), augmented reality (AR), and generative Artificial Intelligence (AI) opens new possibilities for capitalizing on VTS methods in medical education. VR and AR can help create immersive museum-like experiences, allowing learners to engage with religious and spiritual artwork and artifacts even if they lack physical access to museums. In addition, recent advancements in generative AI—such as GPT-4 and easily accessed, responsive video generation—present opportunities for personalized educational content and simulated discussions. At the same time, arts and humanities-based methods can encourage students to reflect on the potential benefits and limitations of using AI tools as well as the uniquely human aspects of patient care.32 Museum-based educational methods, whether conducted in the museum or elsewhere, may ultimately help support core competencies in medical education, provide spiritual care training, and encourage students to reflect on the meaning of religion and spirituality in their personal and professional lives.

    Abbreviations

    VTS, Visual Thinking Strategies; VR, Virtual Reality; AI, Augmented Reality; AI, Artificial Intelligence.

    Disclosure

    Dr. Chisolm is the Director of the Paul McHugh Program for Human Flourishing, through which her work is supported. She also receives compensation for serving as a coach in a Harvard online CME course on VTS. The authors report no other conflicts of interest in this work.

    References

    1. Lipka M, Gecewicz C. More Americans now say they’re spiritual but not religious. Pew Research Center; 2017. Available from: https://www.pewresearch.org/short-reads/2017/09/06/more-americans-now-say-theyre-spiritual-but-not-religious/. Accessed January 12, 2024.

    2. Koenig HG, McCullough ME, Larson DB. Handbook of Religion and Health. 1st ed. Oxford, UK: Oxford University Press; 2001.

    3. Hill PC, Pargament KI, Hood RW, et al. Conceptualizing religion and spirituality: points of commonality, points of departure. J Theory Soc Behav. 2000;30(1):51–77. doi:10.1111/1468-5914.00119

    4. Nissen RD, Viftrup DT, Hvidt NC. The process of spiritual care. Front Psychol. 2021;12:674453.

    5. Crozier D, Greene A, Schleicher M, Goldfarb J. Teaching spirituality to medical students: a systematic review. J Health Care Chaplain. 2022;28(3):378–399. doi:10.1080/08854726.2021.1916332

    6. Jones KF, Paal P, Symons X, Best MC. The content, teaching methods and effectiveness of spiritual care training for healthcare professionals: a mixed-methods systematic review. J Pain Sympt Manage. 2021;62(3):e261–e278. doi:10.1016/j.jpainsymman.2021.03.013

    7. Herschkopf M, Jafari N, Puchalski C. Religion and spirituality in medical education. In: Balboni M, Peteet J, editors. Spirituality and Religion Within the Culture of Medicine: From Evidence to Practice. New York, NY: Oxford Academic; 2017.

    8. Chow HHE, Chew QH, Sim K. Spirituality and religion in residents and inter-relationships with clinical practice and residency training: a scoping review. BMJ Open. 2021;11(5):e044321. doi:10.1136/bmjopen-2020-044321

    9. Religious Landscape Study. Pew research center’s religion & public life project. Available from: https://www.pewresearch.org/religious-landscape-study/database/. Accessed February 28, 2025.

    10. Ambrose AJH, Lin SY, Chun MBJ. Cultural competency training requirements in graduate medical education. J Grad Med Educ. 2013;5(2):227–231. doi:10.4300/JGME-D-12-00085.1

    11. Curtis E, Jones R, Tipene-Leach D, et al. Why cultural safety rather than cultural competency is required to achieve health equity: a literature review and recommended definition. Int J Equity Health. 2019;18(1):174. doi:10.1186/s12939-019-1082-3

    12. So N, Price K, O’Mara P, Rodrigues MA. The importance of cultural humility and cultural safety in health care. Med J Aust. 2024;220(1):12–13. doi:10.5694/mja2.52182

    13. Stubbe DE. Practicing cultural competence and cultural humility in the care of diverse patients. FOC. 2020;18(1):49–51. doi:10.1176/appi.focus.20190041

    14. Gaufberg E, Batalden M. The third thing in medical education. Clin Teach. 2007;4(2):78–81. doi:10.1111/j.1743-498X.2007.00151.x

    15. Gaufberg E, Olmsted MW, Bell SK. Third things as inspiration and artifact: a multi-stakeholder qualitative approach to understand patient and family emotions after harmful events. J Med Humanit. 2019;40:489–504. doi:10.1007/s10912-019-09563-z

    16. Palmer PJ. A Hidden Wholeness: The Journey Toward an Undivided Life. San Francisco, CA: Jossey-Bass; 2009.

    17. Gaufberg E, Williams R. Reflection in a museum setting: the personal responses tour. J Grad Med Educ. 2011;3(4):546–549. doi:10.4300/JGME-D-11-00036.1

    18. Gahtan MW. Museums and exhibitions: overview and history. In: Oxford Research Encyclopedia of Religion. Oxford University Press; 2022.

    19. Gahtan MW. Exhibitions and displays of religious art. In: Oxford Research Encyclopedia of Religion. Oxford University Press; 2022.

    20. Teeter E. Religion and ritual. In: Hartwig MK, editor. A Companion to Ancient Egyptian Art. John Wiley & Sons, Ltd; 2014:328–343.

    21. Greene M. Teaching in a moment of crisis: the spaces of imagination. The New Educator. 2005;1:77–80. doi:10.1080/15476880590934326

    22. Robinson C. Museums and Emotions. J Mus Educ. 2021;46(2):147–149. doi:10.1080/10598650.2021.1922987

    23. Weil SE. From being about something to being for somebody: the ongoing transformation of the American museum. Daedalus. 1999;128(3):229–258.

    24. Chisolm MS, Kelly-Hedrick M, Wright SM. How visual arts-based education can promote clinical excellence. Acad Med. 2021;96(8):1100. doi:10.1097/ACM.0000000000003862

    25. Cerqueira AR, Alves AS, Monteiro-Soares M, et al. Visual Thinking Strategies in medical education: a systematic review. BMC Med Educ. 2023;23(1):536. doi:10.1186/s12909-023-04470-3

    26. Kagan HJ, Kelly-Hedrick M, Benskin E, Wolffe S, Suchanek M, Chisolm MS. Understanding the role of the art museum in teaching clinical-level medical students. Med Educ Online. 2021;27(1):2010513. doi:10.1080/10872981.2021.2010513

    27. Ferrara V, Shaholli D, Iovino A, et al. Visual thinking strategies as a tool for reducing burnout and improving skills in healthcare workers: results of a randomized controlled study. J Clin Med. 2022;11(24):7501. doi:10.3390/jcm11247501

    28. Kagan HJ, Yenawine P, Duke L, Stephens MB, Chisolm MS. Visual thinking strategies and the peril of ‘see one, do one, teach one’. Int Rev Psychiatry. 2023;35(7–8):663–667. doi:10.1080/09540261.2023.2276377

    29. Kelly-Hedrick M, Chugh N, Williams R, Smyth Zahra F, Stephens M, Chisolm MS. The online “personal responses tour”: adapting an art museum-based activity for a virtual setting. Acad Psychiatry. 2022;46(4):510–514. doi:10.1007/s40596-021-01505-z

    30. Stouffer K, Kagan HJ, Kelly-Hedrick M, et al. The role of online arts and humanities in medical student education: mixed methods study of feasibility and perceived impact of a 1-week online course. JMIR Med Educ. 2021;7(3):e27923. doi:10.2196/27923

    31. Kim K, Manohar S, Kalkat M, Iuliano K, Chisolm MS. Museum-based education in health professions learning: a 5-year retrospective. Perspect Med Educ. 2024;13(1):585–591. doi:10.5334/pme.1448

    32. Agarwal G, Yenawine P, Manohar S, Chisolm MS. Implementing a visual thinking strategies program in health professions schools: an AMEE guide for health professions educators: AMEE guide no. 179. Med Teach. 2025:1–10. doi:10.1080/0142159X.2025.2458287

    Continue Reading