NT-proBNP serves as a prognostic marker for adverse outcomes in severe immune checkpoint inhibitor-associated myocarditis
Background Immune checkpoint inhibitors (ICIs) have transformed cancer treatment but carry risks of rare, life-threatening immune-related adverse events, particularly myocarditis. Prognostic biomarkers and optimal management strategies for severe ICI-associated myocarditis remain poorly defined. Methods This single-center retrospective cohort study analyzed 71 patients diagnosed with ICI-associated myocarditis among 7,157 ICI-treated individuals at a tertiary center (August 2018–August 2024). Myocarditis severity was graded per American Society of Clinical Oncology (ASCO) guidelines. Cardiac biomarkers, including NT-proBNP and troponin T, were assessed. Binary logistic regression identified predictors of mortality. Treatment protocols and immunotherapy rechallenge outcomes were evaluated. Results Severe myocarditis (Grades 3–4) occurred in 33 patients (46.5%), with an overall mortality rate of 54.5% in this subgroup (18/33). NT-proBNP levels were significantly elevated in fatal cases versus survivors (median: 13,804 vs. 4,050 pg/mL; P < 0.001) and independently predicted mortality risk (odds ratio (OR) 4.3, 95% confidence interval (CI) [1.2–21.9]; P = 0.023). A multimodal regimen combining plasmapheresis with high-dose corticosteroids, intravenous immunoglobulin, and mycophenolate mofetil was associated with improved survival. Among nine patients rechallenged with immunotherapy, seven (77.8%) tolerated subsequent cycles without recurrent immune toxicity, while two with prior Grade 2 myocarditis experienced symptom recurrence. Discussion Elevated NT-proBNP emerges as a critical prognostic marker for risk stratification in severe ICI-associated myocarditis. Immunotherapy rechallenge appears feasible in select patients but warrants caution in those with prior moderate-grade myocarditis. These findings advocate for biomarker-guided escalation of therapies and shared decision-making frameworks to balance oncologic efficacy with cardiovascular safety.
He et al. (2025) NT-proBNP serves as a prognostic marker for adverse outcomes in severe immune checkpoint inhibitor-associated myocarditis. PeerJ 13:e20020 https://doi.org/10.7717/peerj.20020
Oropharyngeal microbiome dysbiosis in esophageal squamous cell carcinoma: taxonomic shifts, metabolic reprogramming, and geographic disparities in a high-incidence cohort
Background Esophageal squamous cell carcinoma (ESCC) is a leading cause of cancer mortality globally, with pronounced geographic disparities in incidence. Emerging evidence links oral microbiome dysbiosis to ESCC pathogenesis, yet comprehensive insights into microbial diversity, taxonomic shifts, and functional alterations in high-risk populations remain limited. Methods Using 16S rRNA amplicon sequencing, we compared the oral microbiome of ESCC patients and healthy controls from a high-incidence region in Northwest China. Alpha and beta diversity metrics, taxonomic composition, and predicted functional pathways were analyzed to identify microbial signatures associated with ESCC. Results ESCC patients exhibited significantly elevated microbial richness (observed amplicon sequence variants (ASVs), Chao1, ACE; p < 0.05) but comparable Shannon/Simpson diversity to controls. Unique amplicon sequence variants (ASVs) were more prevalent in ESCC samples, and principal component analysis confirmed distinct community structures (p < 0.05). Taxonomically, Streptococcus and Neisseria dominated both groups, but ESCC patients showed enrichment of Gemella (p = 0.0003) and Corynebacterium (p < 0.00001), alongside depletion of Prevotella_7 (p = 0.0002) and Moraxella (p < 0.001). Functional profiling revealed upregulated amino acid metabolism (e.g., beta-alanine and valine degradation) and downregulated carbohydrate metabolism in ESCC-associated microbiota. Conclusion This study uncovers unique oral microbial signatures in ESCC patients from a high-incidence region, characterized by increased richness, taxon-specific shifts, and metabolic reprogramming favoring amino acid catabolism. These findings highlight the potential of microbial biomarkers for ESCC detection and provide mechanistic insights into microbiome-driven carcinogenesis. The geographic specificity of the cohort underscores the urgency of tailored interventions in high-risk populations and advances our understanding of microbial contributions to esophageal cancer.
Liu et al. (2025) Oropharyngeal microbiome dysbiosis in esophageal squamous cell carcinoma: taxonomic shifts, metabolic reprogramming, and geographic disparities in a high-incidence cohort. PeerJ 13:e20009 https://doi.org/10.7717/peerj.20009
The 12-LOX/12-HETE/GPR31 metabolic pathway promotes tumor-associated macrophage M2 polarization mediated pancreatic cancer development
Background The 12-lipoxygenase (12-LOX)/12-hydroxyeicosatetraenoic acid (12-HETE) pathway is associated with various tumors. M2 macrophages in the tumor microenvironment promote tumorigenesis and progression. However, the role of the 12-LOX/12-HETE/G protein-coupled receptor 31 (GPR31) metabolic pathway and its relationship with M2 macrophages remains unclear in pancreatic cancer (PC). Methods The expression levels of 12-LOX, GPR31, and 12-HETE were detected in PC and mouse PC models using western blot and enzyme-linked immunosorbent assays (ELISA). In vivo and in vitro experiments were conducted using the 12-LOX inhibitor ML355 to investigate the role of the 12-LOX/12-HETE/GPR31 metabolic pathway in M2 macrophage polarization and tumor progression through flow cytometry, reverse transcription polymerase chain reaction (RT-PCR), 5-Ethynyl-20-deoxyuridine (EdU) assays, and Transwell experiments. Results The 12-LOX/12-HETE/GPR31 metabolic pathway is expressed actively in PC. Inhibition of 12-LOX in a mouse model of pancreatic cancer suppressed the expression of this metabolic pathway, retarded tumor growth, and reduced the polarization of macrophages towards the M2 type. In vitro, co-culturing PC cell line PANC-1 with macrophages and selectively inhibiting 12-LOX influenced the proliferation, migration, and invasion of PC cells. Inhibiting 12-LOX did not suppress the function of individual PC cells, but it inhibited the development of PC cells co-cultured with macrophages. Moreover, inhibiting 12-LOX reduced the co-cultured M2 macrophages. Conclusion This study, through in vivo and in vitro experiments, reveals that the 12-LOX/12-HETE/GPR31 metabolic pathway affects the growth, migration, and invasion of PC by modulating M2 macrophage polarization patterns.
Yang et al. (2025) The 12-LOX/12-HETE/GPR31 metabolic pathway promotes tumor-associated macrophage M2 polarization mediated pancreatic cancer development. PeerJ 13:e19963 https://doi.org/10.7717/peerj.19963
The BAF53A-BACH1-GCLM axis regulates glutathione metabolism and enhances ferroptosis resistance in esophageal squamous cell carcinoma
Objective Esophageal squamous cell carcinoma (ESCC), a highly lethal malignancy, exhibits poor survival rates and limited treatment options. Ferroptosis, a regulated form of cell death driven by lipid peroxidation, emerges as a potential therapeutic target. However, the mechanisms suppressing ferroptosis in ESCC remain poorly understood. Methods Short hairpin RNA (shRNA) was employed to knock down BAF53A and BACH1 in ESCC cell lines, followed by assessments of cell proliferation, colony formation, and ferroptosis sensitivity. Glutathione (GSH) metabolism was evaluated by measuring GSH/GSSG and NADP+/NADPH ratios, reactive oxygen species (ROS) levels, and lipid peroxidation through flow cytometry and fluorescence imaging. Molecular interactions were evaluated using co-immunoprecipitation and chromatin immunoprecipitation sequencing (ChIP-seq) to identify transcriptional targets of the BAF53A-BACH1 complex. Results BAF53A was elevated in ESCC, and its depletion impaired cell proliferation and colony formation ability of cells. Knockdown of BAF53A disrupted GSH metabolism, leading to increased ROS levels, reduced GSH/GSSG and NADP+/NADPH ratios, and enhanced ferroptosis sensitivity. Mechanistically, BAF53A collaborated with BACH1 to transcriptionally activate glutamate-cysteine ligase modifier subunit (GCLM), a key enzyme in GSH biosynthesis. Overexpression of GCLM restored redox balance and cell viability in BAF53A- or BACH1-silenced cells. Conclusions The BAF53A-BACH1-GCLM axis constitutes a novel egulatory pathway that integrates chromatin remodeling, transcriptional regulatione, and ferroptosis resistance in ESCC. Targeting this axis may offer a promising approach to exploit metabolic vulnerabilities and enhance ferroptosis sensitivity in ESCC treatment.
Jiang et al. (2025) The BAF53A-BACH1-GCLM axis regulates glutathione metabolism and enhances ferroptosis resistance in esophageal squamous cell carcinoma. PeerJ 13:e20156 https://doi.org/10.7717/peerj.20156
The association between ADIPOQ gene variants (rs266729, rs2241766, rs1501299) and acute myocardial infarction in Vietnamese patients with type 2 diabetes mellitus
Background Type 2 diabetes mellitus (T2DM) significantly increases the risk of acute myocardial infarction (AMI). Polymorphisms in the ADIPOQ gene, which encodes the adiponectin hormone, are believed to influence cardiovascular disease risk. This study aims to evaluate the association between three ADIPOQ gene variants—rs266729, rs2241766, and rs1501299—and the risk of AMI in Vietnamese patients with T2DM. Methods A case–control study was conducted from January 2023 to June 2024 at the University Medical Center, Ho Chi Minh City. The study included 275 T2DM patients with a history of AMI (case group) and 275 T2DM patients without AMI (control group). Participants were matched by age, gender, smoking status, and duration of T2DM to control for potential confounding factors. DNA was extracted from peripheral blood samples, and the three polymorphisms were genotyped using Sanger sequencing. Results Significant associations were found between ADIPOQ gene polymorphisms and AMI risk. The G/G genotype of rs266729 and the T/T genotype of rs1501299 were associated with a reduced risk of AMI (OR = 0.45, 95% CI [0.23–0.89], p = 0.021; OR = 0.45, 95% CI [0.22–0.90], p = 0.025, respectively). In contrast, the T/G genotype of rs2241766 was significantly associated with an increased risk of AMI (OR = 2.07, 95% CI [1.44–2.97], p < 0.001). Conclusion This study suggests that rs266729 and rs1501299 may have protective effects, while rs2241766 may increase the risk of AMI in Vietnamese patients with T2DM. These findings highlight the importance of further research into adiponectin levels and long-term monitoring, and support the potential use of genetic variants in personalized cardiovascular risk management for patients with T2DM.
Le et al. (2025) The association between ADIPOQ gene variants (rs266729, rs2241766, rs1501299) and acute myocardial infarction in Vietnamese patients with type 2 diabetes mellitus. PeerJ 13:e20145 https://doi.org/10.7717/peerj.20145
Optimizing legume green manure applications for enhanced forage sorghum-sudangrass performance and soil property improvements
Background Legume-based green manuring is an environmentally sustainable and economically viable approach that enhances soil fertility by improving organic matter content, facilitating biological nitrogen fixation, and stimulating microbial activity. These benefits collectively reduce reliance on mineral fertilizers, which are associated with environmental degradation. While the soil-enhancing properties of legumes are well-established, limited information is available regarding how the timing of green manure incorporation affects the growth performance and forage quality of subsequent crops. This study aimed to assess the effects of incorporating different legume species at various phenological stages on the growth, yield, and forage quality of sorghum × sudangrass under Mediterranean climatic conditions. Methods A field experiment was conducted from 2022 to 2024 at Aydın Adnan Menderes University, Türkiye, using a randomized complete block design with three replicates. Eleven treatments were implemented, consisting of three legume species (common vetch, narbon vetch, and forage pea) incorporated at three distinct phenological stages (pre-flowering, 10% flowering, and full flowering), along with unfertilized and fertilized controls. Following incorporation, sorghum × sudangrass was sown without additional fertilization, except in the fertilized control. Key response variables included fresh forage yield (FFY), hay yield (HY), plant height, leaf number, leaf area index (LAI), chlorophyll content (SPAD), and crude protein yield (CPY). Soil organic matter and total nitrogen levels were also measured before and after treatments. Results Green manure application significantly enhanced the growth, biomass accumulation, and forage quality of sorghum × sudangrass. The incorporation of common vetch and narbon vetch at the 10% flowering stage led to up to a 50% increase in fresh forage yield compared to the unfertilized control. Forage pea maintained consistent yield performance across all incorporation stages. Improvements in soil organic matter and nitrogen content were particularly notable in legume-amended plots. Furthermore, the highest SPAD and LAI values were recorded at early flowering stages, suggesting enhanced photosynthetic efficiency. These findings underscore the effectiveness of legume green manuring, particularly with optimal incorporation timing, as a sustainable strategy to improve soil health, forage productivity, and resource-use efficiency in Mediterranean agroecosystems.
Kara (2025) Optimizing legume green manure applications for enhanced forage sorghum-sudangrass performance and soil property improvements. PeerJ 13:e20137 https://doi.org/10.7717/peerj.20137
Reconstruction of masked sequences via inverse mapping of incomplete information natural vectors
Alignment-free embedding methods, which map biological sequences into a fixed-dimensional space using mathematical techniques, hold significant value in biology. A key challenge in this field is constructing an inverse mapping to recover sequences from embedded vectors. The natural vector approach uniquely provides a theoretical one-to-one correspondence between sequences and high-order natural vectors, but reconstructing sequences from lower-order vectors remains unsolved. Moreover, when sequences contain masked regions, extracting features and constructing the inverse mapping to restore the original sequence, including the masked parts, becomes even more challenging. In this article, we define incomplete information natural vectors for masked sequences and develop a long short-term memory model that achieves over 99.9% accuracy in reconstructing unmasked positions in original sequences on SARS-CoV-2 and HIV-1 datasets, while also providing predictions for masked sites that significantly outperform random prediction. Our model can robustly handle sequences with varying masked nucleotides. Overall, our approach expands the scope of alignment-free embedding methods by enabling bidirectional conversion and addressing challenges posed by incomplete information.
Ding et al. (2025) Reconstruction of masked sequences via inverse mapping of incomplete information natural vectors. PeerJ 13:e20126 https://doi.org/10.7717/peerj.20126
Proteomic landscape of porcine induced neural stem cell reprogramming and differentiation
Background Direct reprogramming of somatic cells into induced neural stem cells (iNSCs) holds strong potential for regenerative medicine, especially in large animal models like pigs, which are crucial for translational and preclinical research. However, the molecular mechanisms underlying porcine fibroblast-to-iNSC reprogramming and subsequent differentiation remain poorly understood at the proteomic level. Methods To map the proteomic landscapes associated with reprogramming and differentiation, we performed unbiased label-free discovery proteomics (nano-LC-MS/MS) and targeted SWATH-MS quantification. Proteomes of porcine tail fibroblasts (PTFs; passage 3), two porcine iNSC lines (piNSCs; VSMUi002-B and VSMUi002-E, passage 20), and their differentiated progeny (piNSCs-NGs; VSMUi002-B-NGs and VSMUi002-E-NGs, representing piNSCs at passage 20 cultured for an additional 14 days under differentiation conditions) were compared. Two previously established piNSC lines (VSMUi002-B and VSMUi002-E), generated via Sendai virus-mediated reprogramming, were used as the cellular models. Results The piNSC lines displayed hallmark neural stem cell (NSC) morphology and expressed canonical markers (PAX6+, SOX2+, NES+, VIM+, OCT4−). Upon differentiation, they generated neuronal and glial cells expressing TUJ1, MAP2, SYP, TH, and GFAP, confirming their multipotency. A total of 4,094 proteins were identified across the three cell states. Multivariate analysis revealed distinct proteomic signatures separating fibroblasts, iNSCs, and their neuronal/glial progeny. The proteomic shift from the fibroblast to the piNSC state was marked by increased expression of stathmin 1 (STMN1), neurofilament light polypeptide (NEFL), aconitate hydratase (ACO2), electron transfer flavoprotein subunit beta (ETFB), fructose-bisphosphate aldolase B (ALDOB), and transketolase (TKT), alongside suppression of actin-related protein 2/3 complex subunit 5 (ARPC5) and LIM domain and actin-binding protein 1 (LIMA1). These shifts indicate a dismantling of the fibroblast cytoskeleton and a broad upregulation of cellular energy and biosynthetic metabolism, reflecting a loss of fibroblast identity and the acquisition of an NSC state. Upon differentiation into piNSCs-NGs, 19 proteins were consistently upregulated. These included neuronal structural proteins (INA, STMN1), cytoskeletal regulators (PFN1), signaling modulators (MBIP), and proteins involved in lysosomal function (NCOA7), cell adhesion (CDHR2), and calcium signaling (ANXA4). Pathway and network analyses highlighted post-transcriptional regulation—particularly involving RNA processing and the RNA exosome complex (e.g., EXOSC3)—as a key feature of differentiation. Conclusion This study provides the first comprehensive proteomic map of piNSC reprogramming and differentiation in a large animal model. Our findings uncover critical regulatory proteins and pathways governing cytoskeletal organization, metabolism, and RNA processing, offering valuable insights into neural fate states. This resource advances the understanding of neural reprogramming in translational models and supports future regenerative and comparative neuroscience efforts.
Ploypetch et al. (2025) Proteomic landscape of porcine induced neural stem cell reprogramming and differentiation. PeerJ 13:e20120 https://doi.org/10.7717/peerj.20120
Exploration of nitrogen use efficiency and root-shoot-soil variation on the stoichiometric characteristics of carbon, nitrogen, phosphorous and potassium for winter wheat under various nitrogen treatments
Background Nitrogen (N) addition has significant effects on grain yield and nutrient use efficiency (NUE) in farmland ecosystems, and the total carbon: nitrogen: phosphorus: potassium (TC:TN:TP:TK) stoichiometry in plants and soil is of great significance for improving plant productivity and nutrition. However, there is still uncertainty regarding the effects of N fertilizers on the stoichiometric characteristics of different root-shoot parts of winter wheat during growth progression. Methods ‘Fengdecunmai5’ wheat was selected as the material for the positioning test field. Three N application treatments (0, 180 and 300 kg ha−1, designated N0, N180 and N300) were designed to study the yield, NUE, and stoichiometric characteristics of TC, TN, TP and TK in each organ at different stages, and to analyze the relationships among them. Results The N180 treatment increased the total nitrogen (TN) content in roots, stems plus leaves and grain at each stage. The TN and TP content in stems plus leaves gradually decreased from jointing to maturity stages, whereas the TN content in grain gradually increased from filling to maturity stages. Under the N180 treatment, TN:TP values significantly increased, by 20.3%, 27.9% and 26.3% compared with N0 treatment, respectively. Moreover, the farmland ecosystem was mainly limited by the TN content. TK content in roots and TC:TN in stems plus leaves was significantly positively correlated with NUE. TN:TK and TN: TP in stems plus leaves were positively correlated with yield, and TN and TK content in stems plus leaves considerably influenced the yield. Conclusions Under N180 treatment, compared with N0 and N300 treatments, wheat plants achieved higher nutrient uptake, and greater yield and NUE. This study contributes to the assessment of plant productivity and the precise optimization of ecosystem stoichiometry in the context of green sustainable agricultural development.
Lv et al. (2025) Exploration of nitrogen use efficiency and root-shoot-soil variation on the stoichiometric characteristics of carbon, nitrogen, phosphorous and potassium for winter wheat under various nitrogen treatments. PeerJ 13:e20101 https://doi.org/10.7717/peerj.20101
Influence of organic, synthetic and biofertilizers on the diversity of cassava rhizosphere microbiome in Northeastern Thailand
Cassava, one of Thailand’s main economic crops, is capable of growing in nearly all soil types. However, continuous monocropping depletes soil nutrients over time. Adopting good agricultural practices can help farmers reduce costs while improving soil fertility. The aim of this study was to compare cassava rhizosphere microbial communities resulting from cultivation under eight different fertilizer treatments, including synthetic, organic, and biological fertilizers, and to identify beneficial microbes that promote cassava growth and yield. The study was conducted at two sites in Northeastern Thailand. Results show that bacterial abundance and species richness (alpha diversity) peaked at 5 months after planting (MAP), showing a significant increase compared to 2 MAP. However, by 10 MAP, alpha diversity began to decline at both sites, Nampong and Seungsang. Among the treatments, the most notable differences in alpha diversity were observed at 5 MAP. At the Nampong site, experimental treatments with chicken manure (T3) and chicken manure combined with other fertilizers (T5, T6, and T8) exhibited significantly higher alpha diversity than did the control (without fertilizer, T1). At the Seungsang site, sole treatment with the full recommended rate of chicken manure (T3), and half of the recommended dose of synthetic fertilizer combined with half the recommended dose of chicken manure (T6) resulted in greater alpha diversity than did swine manure extract application (T4), half of the recommended dose of synthetic fertilizer combined with half of the recommended dose of swine manure extract (T7), and chicken manure application combined with stalk inoculation with plant growth-promoting rhizobacteria (PGPRs) (T8). Since T3 and T8 had the most significant impact on microbial abundance and diversity, as well as cassava growth and yield, the predominant bacteria in these treatments were identified as key targets. A total of eight target bacterial genera were identified: Pseudomonas, Tumebacillus, Lysinibacillus, Paenibacillus, Dongia, Acidibacter, Sphingomonas, and Bacillus. Among them, Tumebacillus was the most notable, as it showed a significant correlation with fresh tuber yield. These beneficial bacteria may serve as key candidates for future biofertilizer production.
Somyong et al. (2025) Influence of organic, synthetic and biofertilizers on the diversity of cassava rhizosphere microbiome in Northeastern Thailand. PeerJ 13:e20085 https://doi.org/10.7717/peerj.20085