ABSTRACT Background Obesity impairs endothelial nitric oxide synthase (eNOS)‐nitric oxide (NO) signaling, triggering chronic inflammation, endothelial dysfunction and defective vasodilation, which drives cardiometabolic diseases. Existing metabolic therapies provide only indirect vascular protection and fail to resolve residual inflammatory‐mediated cardiovascular risk. As an anti‐inflammatory interleukin 12 (IL‐12) family cytokine, interleukin 35 (IL‐35) is suppressed in obesity and exerts vascular protective effects under inflammatory conditions, yet its function and underlying mechanisms in obesity‑related endothelial vasodilator dysfunction remain undefined. Materials and Methods An obese mouse model was established by 8‐week high‐fat diet (HFD) feeding, followed by 4‐week intraperitoneal administration of recombinant mouse IL‐35. Aortic‐ring assays evaluated vasodilatory function. Palmitic acid (PA)‐treated human umbilical vein endothelial cells (HUVECs), recapitulate lipotoxic endothelial injury, were treated with recombinant human IL‐35. Western blotting, immunofluorescence, 4‐amino‐5‐methylamino‐2′,7′‐difluorofluorescein diacetate (DAF‐FM DA) detection and immunoprecipitation‐mass spectrometry (IP‐MS) were applied to detect IL‐35‐subunit (Epstein‐Barr virus‐induced gene 3 [EBI3] and interleukin‐12 subunit alpha [IL‐12A]) levels, eNOS Ser1177 phosphorylation, phosphorylation of mitogen‐activated protein kinase/extracellular signal‐regulated kinase (MAPK/ERK) signaling molecules (cellular rapidly accelerated fibrosarcoma [cRAF], mitogen‐activated protein kinase kinase [MEK], and ERK) and signal transducer and activator of transcription 3 (STAT3) activation, NO production, and eNOS‐binding partners. Rescue assays were performed using Raf or ERK specific inhibitors and STAT3 small interfering RNA (siRNA). Results HFD mice displayed reduced aortic endothelial EBI3 expression, impaired endothelium‐dependent vasorelaxation, decreased eNOS Ser1177 phosphorylation and lowered NO production; all these phenotypes were rescued by exogenous IL‐35. IP‐MS and immunoblotting demonstrated that IL‐35 strengthened eNOS‐ERK1 interaction and restored PA‐repressed phosphorylation of cRAF, MEK, ERK and eNOS Ser1177, which were abrogated by Raf or ERK blockade. Furthermore, IL‐35 recovered STAT3 phosphorylation suppressed by PA, while STAT3 knockdown blunted IL‐35‐driven MAPK/ERK activation, eNOS phosphorylation and subsequent NO production. Conclusion IL‐35 alleviates obesity‐induced impairment of endothelium‐dependent vasodilation via the STAT3‐MAPK/ERK‐eNOS axis, which elevates eNOS Ser1177 phosphorylation and NO synthesis. This work supports IL‐35 as a prospective therapeutic candidate for obesity‐associated vascular complications.
ABSTRACT Background Chronic airway inflammation can be induced by both chemical and environmental factors. Airway inflammation can be induced through different immune pathways, including allergic Th2‐dependent activation of eosinophils and mast cells, Th1/Th17‐dependent neutrophil recruitment, or a combination of both mast cell and neutrophil recruitment, that is extremely difficult to treat. An environmental factor that we have shown to induce neutrophil‐mediated lung inflammation is cyanobacterium lipopolysaccharide (LPS). The study has two main objectives: first to determine whether TLR4 is required for cyanobacterium LPS‐induced lung inflammation, given our prior findings that it signals differently from other LPS types, and second to test if cyanobacterium LPS exacerbates lung inflammation in a Th2‐dependent mouse model of airway hypersensitivity. Methods LPS from the cyanobacterium strain, Brasilonema sp., was purified and used for both in vivo and in vitro assays. For in vitro assays, the TLR4 inhibitor, SPA‐4, was assessed in its ability to block Brasilonema sp.‐dependent IL‐6 and IL‐8 induction in lung epithelial cell lines, and in vivo, the ability of Brasilonema sp. LPS to induce lung inflammation was assessed in TLR4‐deficient mice in comparison to controls. Finally, Th2‐driven lung inflammation was established using ovalbumin and then these mice were exposed to Brasilonema sp. LPS intranasally to assess if Brasilonema sp. LPS resulted in the exacerbation of Th2‐dependent airway inflammation. Results Our results indicate that Brasilonema sp. LPS stimulates the production of cytokines both in vitro and in vivo in a TLR4‐dependent manner, since the addition of the SPA‐4 peptide blocks Brasilonema sp. LPS‐dependent increases in IL‐6 and IL‐8 in vitro and TLR4‐deficient mice were refractory to Brasilonema sp. LPS‐mediated induction of lung inflammation in vivo. Additionally, both E. coli and Brasilonema sp. LPS exacerbated airway inflammation in a Th2‐dependent model of lung inflammation. However, Brasilonema sp. LPS exacerbated Th2‐dependent lung inflammation without significant neutrophil infiltration. Conclusions Despite the altered signaling through TLR4 by cyanobacterium LPS, these data demonstrate that lung inflammation induced by LPS from the cyanobacterium strain, Brasilonema sp., requires TLR4 and exacerbates Th2‐dependent lung inflammation. Therefore, novel interventions that target TLR4 may be beneficial in the treatment of LPS‐induced lung inflammation from environmental exposure.
ABSTRACT Background Allergic Rhinitis (AR) is a widespread condition that significantly affects the quality of life (QOL) of those affected globally. It contributes to sleep disturbances, fatigue, poor concentration, irritability, and can worsen symptoms in individuals with coexisting asthma. This study examines the impact of AR on QOL in adults, with specific attention to sex differences. Materials and Methods A cross‐sectional online survey using a purpose‐designed, non‐validated questionnaire was distributed via social media using convenience sampling to 1572 Jordanian adults. The questionnaire collected data on demographics, AR symptoms, triggers, and the impact of AR on mental health, social functioning, and daily life. Data were analyzed using descriptive statistics and Chi‐square tests. Multivariable logistic regression was used to adjust for potential confounders, and Benjamini–Hochberg false discovery rate (FDR) correction was applied to account for multiplicity. Results A total of 1572 respondents participated; 683 (43.4%) had physician‐diagnosed AR. AR was reported by 157/419 males (37.5%) and 526/1153 females (45.6%); this sex difference was statistically significant ( χ 2 = 8.31, p = 0.004; OR = 1.40, 95% CI 1.11–1.76). Most patients lived in urban areas (69.4%). While sneezing and rhinorrhoea were common across sexes, females reported significantly higher rates of nasal itching/congestion (55.5% vs. 43.9%, p = 0.011), headaches (61.0% vs. 43.9%, p < 0.001), and increased stress/anxiety (53.4% vs. 42.7%, p = 0.018, not surviving false discovery rate (FDR) correction). In contrast, snoring was more prevalent in males (40.1% vs. 24.1%, p < 0.001). Regarding QOL, AR severely disrupted sleep (80.7%) and concentration (66.2%) for both sexes. However, females were significantly more likely to report irritability/emotional distress (65.4% vs. 55.4%, p = 0.023), prefer social isolation during symptoms (59.3% vs. 47.8%, p = 0.010), and experience limitations in activities outside the home (55.1% vs. 45.9%, p = 0.041). After adjustment, female sex remained significantly associated with irritability/emotional distress (aOR = 1.50, 95% CI 1.02–2.22, p = 0.039) and preference for social isolation (aOR = 1.53, 95% CI 1.05–2.23, p = 0.028). After Benjamini–Hochberg FDR correction, nasal itching/congestion, snoring, and headache remained statistically significant in symptom analyses; no QOL items survived FDR correction, and these findings should be considered exploratory. Asthma was reported by 80/683 (11.7%) of AR patients, more frequently among males than females (19.7% vs. 9.3%, p < 0.001). Sensitivity analyses showed that the association between female sex and social isolation was robust to adjustment for or exclusion of asthma, whereas the association with irritability was attenuated after excluding participants with asthma. Conclusion These findings suggest AR's substantial impact on daily functioning and mental well‐being, with signals of greater psychosocial burden among women. Improved understanding of sex‐specific challenges can support more effective prevention, diagnosis, and management strategies, with tailored strategies to address the distinct symptom burden and QOL impairments experienced by female patients. These findings may provide preliminary insights into the burden of AR in Jordan and should be confirmed in representative, validated surveys.
ABSTRACT Background Allergic rhinitis (AR) represents a highly prevalent inflammatory disorder of the nasal mucosa, but molecular indicators capable of capturing its disease‐specific transcriptional features remain insufficiently defined. This study aimed to identify cell adhesion‐related diagnostic biomarkers for AR and to investigate their immune relevance and molecular heterogeneity. Methods Publicly available AR microarray datasets were analyzed, with GSE75011 serving as the discovery dataset and GSE46171 and GSE40037 used to test the reproducibility of the diagnostic model. A predefined adhesion‐related gene set was curated from MSigDB to provide the biological search space for biomarker screening. Genes showing disease‐associated expression changes were overlapped with this adhesion gene set to obtain AR‐associated adhesion genes. Their biological functions and interaction structure were examined through enrichment analysis, protein interaction mapping, and module extraction. Diagnostic candidates were prioritized by integrating three complementary feature‐selection strategies: least absolute shrinkage and selection operator regression, random forest modeling, and support vector machine‐recursive feature elimination. The selected markers were then assessed for diagnostic discrimination, individualized prediction, calibration performance, and potential clinical net benefit. In addition, immune infiltration, upstream regulatory networks, candidate compounds, and non‐negative matrix factorization‐based molecular subtypes were explored. Results A total of 38 differentially expressed adhesion‐related genes were identified in AR, and 13 hub genes were extracted from the key protein–protein interaction module. Integrated machine learning ultimately prioritized TNFSF14 and CD83 as diagnostic biomarkers. The combined TNFSF14/CD83 model showed strong diagnostic separation in GSE75011, with an area under the curve (AUC) of 0.968, and maintained external performance in GSE46171 and GSE40037, with AUC values of 1.000 and 0.758, respectively. Both biomarkers were linked to altered immune‐cell features, particularly T‐cell‐related signatures. Using the two‐marker expression pattern, AR samples could be separated into two exploratory molecular patterns that showed differences in immune profiles and enriched biological processes. Conclusions TNFSF14 and CD83 may represent promising candidate diagnostic biomarkers associated with AR‐related immune alterations. These findings also reveal immune heterogeneity within AR and provide a bioinformatics‐based framework for biomarker discovery and molecular stratification.
ABSTRACT Background Gastric cancer remains a major challenge in public health, with low early diagnosis rates and poor prognosis for advanced patients. Heat shock proteins (HSPs) are highly conserved molecular chaperones that maintain proteostasis and promote cell survival under stress. Aberrantly elevated HSPs in gastric cancer are closely linked to tumor initiation, invasion, metastasis, and poor prognosis. This review aims to comprehensively synthesize the latest advances concerning HSPs in gastric cancer across three core dimensions: their utility as diagnostic/prognostic biomarkers, their regulatory mechanisms in key signaling pathways and the tumor microenvironment (TME), and their viability as therapeutic targets. Materials and Methods This review is structured as a narrative synthesis of the literature. A structured search was performed across PubMed, EMBASE, Web of Science Core Collection, and Cochrane Library, from their inception to March 2026. The search strategy combined keywords and MeSH terms related to three domains: (1) HSPs; (2) gastric cancer; and (3) therapeutics/inhibitors. Two authors independently screened titles and abstracts, and disagreements were resolved through discussion or consultation with a third author. Observational human studies, experimental studies (in vitro and in vivo), and preclinical animal models relevant to the three thematic pillars were included. Reference lists of included papers and relevant review articles were also manually searched. Given the broad scope and qualitative, integrative nature of this review, no formal quantitative meta‐analysis or standardized risk‐of‐bias assessment was performed. Instead, findings were synthesized thematically, emphasizing critical appraisal of conflicting data, identification of knowledge gaps, and discussion of translational barriers. Results HSPs exhibit heterogeneous expression in gastric cancer tissues, which is associated with tumor differentiation, lymph node metastasis, and progression. HSP70 expression correlates with poor differentiation, high invasiveness, intestinal‐type gastric cancer, and lymphovascular invasion, and is involved in proliferation, migration, anti‐apoptosis, and chemoresistance; inhibition of HSP70 enhances therapeutic sensitivity. HSP90ab1 interacts with LRP5, inhibits its ubiquitin‐mediated degradation, activates AKT and Wnt/β‐catenin signaling, and promotes epithelial–mesenchymal transition and invasive metastasis. Plasma HSP90α is elevated in gastric cancer patients and correlates with CEA, CA125, CA153, CA199, and immune cells, as well as with metastatic stage. HSP27 is elevated in gastric cancer tissues, promotes tumor growth by inhibiting PANoptosis, and is associated with progression and poor prognosis. HSPs also participate in TME remodeling by regulating cancer‐associated fibroblasts, immune cells, vascular endothelial cells, and the extracellular matrix. They can both activate anti‐tumor immunity and promote immunosuppression and immune evasion, and are involved in multidrug resistance. Mechanistically, HSPs interact with pathways such as PI3K/Akt, MAPK, and TGF‐β, showing context‐dependent and biphasic regulatory roles. HSP inhibitors, especially HSP90 inhibitors such as ganetespib, have shown preclinical and clinical activity in gastric cancer, but single‐agent efficacy is limited by toxicity, poor tumor penetration, and HSF1‐driven compensatory upregulation of HSP70, HSP40, and HSP27. Combination with chemotherapy, immunotherapy, photothermal/photodynamic therapy, and nanocarrier‐based delivery has shown potential to improve efficacy. Dual targeting of HSP70/HSF1 and individualized combination regimens are proposed future directions. Discussion HSPs are key molecules in the initiation and progression of gastric cancer. Their abnormal expression reflects tumor biological characteristics and provides new avenues for clinical diagnosis and treatment. The mechanisms of HSPs in gastric cancer are complex, involving stress responses, protein folding/repair, signaling pathway regulation, and immune evasion, which determine their critical roles in the TME. As biomarkers, HSPs have shown sensitivity and specificity in early diagnosis and prognosis, but clinical translation still faces challenges such as inhibitor selection, protection of normal cells, and drug resistance. Future research requires multidisciplinary collaboration to resolve existing controversies, clarify molecular mechanisms, and develop efficient and safe targeted drugs, so as to improve the diagnosis and treatment outcomes of gastric cancer and enhance patient survival and quality of life.
