Search Results (426)

Background/Objectives: Lipid nanoparticles (LNPs) have emerged as crucial vehicles for messenger RNA (mRNA) applications in antitumor therapy. Combining LNPs with stimulator of interferon genes (STING) activation holds promise for treating “cold” tumors such as pancreatic cancer. However, two major challenges remain: inefficient mRNA escape from endosomes and STING pathway suppression in immunosuppressive tumor microenvironments. Methods: To improve endosomal escape, we developed a novel pH-responsive PEGylated lipid (Ben-mPEG₂₀₀₀) for mRNA-LNP preparation while using commercial Man-mPEG₂₀₀₀ for dendritic cell (DC)-targeted delivery of LNPs; to alleviate suppression of the STING pathway in the tumor microenvironment and activate immune responses, STING-R283S mRNA was encapsulated into LNPs, ultimately resulting in DC-targeted/pH-responsive LNPs loaded with STING-R283S mRNA for pancreatic cancer immunotherapy research. Results: After pH-responsive cleavage, Ben-mPEG₂₀₀₀ not only enhanced the positive charge of LNPs through the exposed protonated amino groups but also eliminated the PEG-induced steric hindrance effect. The combination of these two effects promoted membrane fusion between LNPs and the endosome, thereby enhancing mRNA translation. As a payload, STING-R283S could further amplify STING signaling in DCs without cytotoxicity to counteract immunosuppression in pancreatic cancer. Conclusions: This engineered LNP platform enhanced mRNA expression and STING activation in DCs, improving immunotherapy outcomes in pancreatic cancer. Full article

Chilling injury is a major problem limiting the postharvest storage and marketability of mango fruit at low temperature. The present study investigated the individual and combined effects of sodium nitroprusside (SNP), L-arginine (Arg) and salicylic acid (SA) on chilling tolerance, regulation of oxidative stress and the postharvest quality of ‘Keitt’ mango fruit stored at 5 ± 1 °C for 28 days followed by 4 days of shelf life at 23 °C. Fruits were pre-treated with 1 mM SNP, 1 mM Arg, 2 mM SA or their binary combinations before storage. The chilling injury, membrane damage, lipid peroxidation, protein oxidation and fruit softening were greatly enhanced by cold storage in untreated fruits. In contrast, all the treatments significantly ameliorated these deteriorative changes, and the combined treatments were superiorly effective. Among these, SNP + Arg was the most effective treatment, which reduced the chilling injury index from 4.05 in control fruits to 1.00 after shelf life, completely inhibiting the incidence of decay and reducing electrolyte leakage and malondialdehyde accumulation by 47.4 and 48.2%, respectively. The same treatment also maintained higher firmness, titratable acidity, visual appearance and ascorbic acid content than untreated fruits. The enhanced chilling tolerance was accompanied by increased antioxidant defense, as SNP + Arg significantly stimulated the activities of superoxide dismutase, catalase and peroxidase, but suppressed the activity of pectin methylesterase. Multivariate analyses, such as PCA, clustered heatmap and integrated stress index, demonstrated a strong negative relationship between oxidative stress markers and antioxidant metabolism. The results showed that combined SNP and Arg treatments enhanced chilling tolerance through increasing antioxidant capacity, preserving membrane integrity, and retarding ripening-related metabolism, which provides an effective way to maintain the postharvest quality of cold-stored mango fruit. Full article

Reusing wastewater in agriculture is essential due to water scarcity but requires treatment technologies that preserve soil biological integrity. This study evaluated the impact of irrigation with wastewater treated by cold atmospheric plasma (CAP; 60 min exposure) on the soil microbiome during Festuca rubra L. and Sinapis alba L. cultivation. The experimental design included various CAP-wastewater dilutions evaluated in two replicates (n = 2), with microbial shifts assessed via 16S rRNA gene sequencing. CAP treatment reduced non-purgeable organic carbon (NPOC) while enriching the water with nitrogen, which significantly stimulated S. alba root growth. Metagenomic analysis confirmed high microbiome stability. Dominant phyla (Proteobacteria and Actinobacteriota) remained stable, and beta-diversity indices showed no statistically significant ecological shifts (R² = 0.420, p = 0.121). Furthermore, CAP-treated irrigation promoted beneficial taxa, specifically the genus Bacillus. These findings demonstrate that CAP wastewater treatment is a safe, environmentally responsible strategy for wastewater reclamation. It successfully supports nutrient cycling and agricultural production without compromising soil microbial homeostasis or health, offering a viable solution aligned with the principles of a circular economy. Full article

Background: Paranasal sinuses and mastoid air cells have been attributed to multiple functions—such as voice resonance, cranial lightening, and pressure regulation—yet their potential role in local thermal homeostasis remains underappreciated. The thermoregulatory hypothesis, first proposed in the mid-twentieth century, was largely abandoned after the mid-century, when anthropological findings of climate-correlated variation seemed contradictory. Hypothesis: We propose that pneumatized skull regions form a three-component craniofacial biothermal system that maintains thermal stability in the ocular vitreous and vestibular endolymph, two avascular, temperature-sensitive structures that lack intrinsic thermoregulatory capacity. This represents a novel integration that explicitly links paranasal and mastoid pneumatization into a coordinated system that protects sensory organs, distinct from previous brain-cooling hypotheses. Mechanism: The system comprises: (1) passive thermal insulation via air spaces, providing ~15-fold greater thermal resistance than bone; (2) active cold protection via mucosal heat delivery (estimated 2–5 W capacity); and (3) active heat dissipation via evaporative cooling (estimated 0.3–0.5 W capacity). This architecture provides asymmetric protection, with cold buffering exceeding heat dissipation by approximately 5- to 15-fold, consistent with thermodynamic constraints and putative evolutionary priorities. Evidence: Preliminary observations consistent with this hypothesis include the anatomical proximity of pneumatized regions to the vitreous and labyrinth, intranasal selective brain cooling studies, and clinical observations after mastoidectomy showing preserved pressure buffering but reduced vestibular thermal insulation under extreme stimulation. Climate-correlated pneumatization patterns are consistent with bidirectional thermal adaptation. Implications: We present five falsifiable predictions that can be tested with thermographic imaging, pharmacological manipulation, and computational modeling. Validation could inform surgical planning, explain postoperative thermal-sensitivity symptoms, and provide evolutionary insights into craniofacial adaptation. Full article

Background/Objectives: Respiratory syncytial virus (RSV), human metapneumovirus (hMPV), and parainfluenza virus type 3 (PIV3) are leading causes of acute respiratory infections in children and the elderly, yet no licensed T-cell vaccines are available. This study aimed to develop multivalent T-cell vaccine candidates against these pathogens using a live attenuated influenza virus (LAIV) vector platform. Methods: Conserved F, N, and M proteins of RSV, hMPV, and PIV3 were identified through multiple sequence alignments. Fragments enriched with experimentally confirmed and predicted T-cell epitopes were selected using the IEDB and NetMHCpan servers. These fragments were assembled into polyepitope immunogenic cassettes, and their selected order was determined by thermodynamic analysis of mRNA secondary structures using the RNAfold Web Server. The selected cassettes were cloned into the neuraminidase (NA) gene of a cold-adapted LAIV vector. Recombinant viruses were rescued by reverse genetics and assessed for replicative fitness in embryonated chicken eggs and MDCK cells, NA enzymatic activity and genetic stability upon serial passaging. Results: Four cassettes were designed for RSV, three for hMPV, and one for PIV3, all containing fragments with multiple T-cell epitopes. Three recombinant viruses of LAIV/RSV type and three of LAIV/hMPV type were successfully rescued, while attempts to recover the remaining recombinant viruses, i.e., LAIV/RSV and LAIV/PIV3, were not successful. All rescued recombinant viruses replicated to titers comparable to the parental LAIV strain and retained the full-length insert for at least eight passages in eggs. Importantly, NA enzymatic activity of the LAIV vector was not compromised by the insertion of the polyepitope T-cell cassettes. Conclusions: We developed a panel of recombinant T cell-based vaccine candidates against RSV and hMPV using the LAIV vector platform. These recombinant viruses encode conserved T-cell epitopes of the target viruses while retaining the biological properties of LAIV strains. Taken together, these characteristics warrant further evaluation of these recombinant viruses in appropriate relevant in vitro models to directly assess their immunogenicity in terms of stimulating a T-cell response against target pathogens. Full article

SIRT2 (Sirtuin 2) is an NAD+-dependent deacetylase that exerts crucial regulatory effects on immune homeostasis and macrophage activation. While chronic cold exposure is a known predisposing factor for pulmonary dysfunction, the precise mechanisms by which SIRT2 potentially modulates lung macrophage polarization under cold stress remains poorly understood. In this study, we evaluated the protective capacity of SIRT2 using both wild-type (WT) and Sirt2-knockout (Sirt2−/−) murine models subjected to chronic cold exposure (4 °C for 3 h daily over 21 days). Our results demonstrated that Sirt2 deficiency significantly exacerbated cold-induced pulmonary histopathological damage and increased the secretion of pro-inflammatory cytokines (TNF-α, IL-1β, and IL-6) (p < 0.05). Furthermore, chronic cold stress triggered a macrophage-centered inflammatory response, a process wherein SIRT2 was found to curtail M1 pro-inflammatory polarization. To further investigate these mechanisms, in vitro experiments were conducted using the mouse alveolar macrophage cell line MH-S. While LPS was utilized as a canonical inflammatory stimulus to mimic the injury environment, SIRT2 overexpression was found to reverse the LPS-induced increase in M1 markers and attenuate inflammatory cytokine secretion. These findings suggest that SIRT2 maintains intracellular homeostasis by modulating macrophage plasticity and plays a protective role in the development of chronic cold stimulus-induced lung injury. Consequently, SIRT2 activation may represent a potential therapeutic pathway for the treatment of environment-related respiratory diseases. Full article

To investigate the effects of water-saving irrigation and different straw retention methods on soil CH₄ and N₂O emissions from paddy fields in cold regions and their potential underlying mechanisms, a field experiment was conducted in Qing’an City, Heilongjiang Province. Two water management regimes were set, combined with four straw retention treatments. The static chamber-gas chromatography method was used to monitor CH₄ and N₂O emission fluxes during the entire rice growth period. Meanwhile, soil pH, oxidation–reduction potential (Eh), dissolved oxygen (DO), and dynamic changes in carbon and nitrogen substrates were measured, and the global warming potential (GWP) and greenhouse gas emission intensity (GHGI) were comprehensively evaluated. The results showed that controlled irrigation significantly increased soil dissolved oxygen content and oxidation–reduction potential. Compared with conventional flooding irrigation, total CH₄ emission decreased by more than 50%, while N₂O emission increased by 1.5–2.5 times, exhibiting an obvious divergent correlation with the two gas emission fluxes. Among different straw retention methods, organic fertilizer returning and direct straw returning significantly promoted CH₄ emission by supplying easily decomposable organic carbon. In contrast, biochar, due to its stable carbon structure and favorable pore properties, inhibited CH₄ emission without significantly stimulating N₂O emission. The treatment of controlled irrigation combined with biochar returning (CB) achieved the lowest global warming potential and greenhouse gas emission intensity at 7230.82 kg CO₂-eq/hm² and 0.8054 kg CO₂-eq/kg, respectively, while maintaining high rice yield. Path analysis based on soil physicochemical properties and emission fluxes further revealed that Eh and DO were significantly negatively correlated with CH₄ emission but positively correlated with N₂O emission. Path inference from flux and substrate data indicated that carbon and nitrogen availability were the key factors limiting the denitrification process. In conclusion, the combined application of controlled irrigation and biochar returning can realize the synergistic effect of stable yield and emission reduction in cold-region paddy fields by improving soil aeration and regulating the transformation of carbon and nitrogen substrates, providing a scientific basis for establishing a green and low-carbon rice production technology system for black soil in cold regions. Full article

Ridge tillage (RC) has been proposed as a water-saving irrigation technique to mitigate greenhouse gas (GHG) emissions from paddy fields. To evaluate its effectiveness under cold-region climatic conditions, a two-year field experiment (2023–2024) was conducted in Northeast China. The study assessed the effects of RC on rice yield, methane (CH₄), nitrous oxide (N₂O), and total GHG emissions (expressed as CO₂e). A no-puddling treatment (NP) was additionally included in 2024. The results showed that compared to conventional cultivation (CK), RC significantly increased the number of effective panicles in 2023 (p < 0.05) but did not significantly affect yield in either year. CH₄ emissions exhibited a double-peak pattern, with peaks at the heading and grain-filling stages; the heading stage contributed the largest part (53.1–69.0%). N₂O emissions showed no distinct seasonal pattern, although N fertilization events stimulated N₂O peak. RC consistently reduced CH₄ emissions, with reductions of 50.8% in 2023 and 71.0% in 2024. NP in 2024 reduced CH₄ emissions by 27.0%. N₂O emissions showed no significant differences among treatments; however, their contribution from fertilization events varied with treatment and year. Total GHG was dominated by CH₄ (>99%). RC significantly lowered GHG and GHGI by 50.7–70.1% and 57.9–73.2% compared to CK, respectively. In conclusion, ridge tillage is an effective practice to reduce CH₄ and GHG emissions while maintaining rice yield in cold-region paddy fields. The large inter-annual variability strongly affects baseline emissions and underscores the needs for multi-year assessments. Full article

Macrophages play a crucial role in health and disease. Currently, reporter mice for tracking alternatively activated macrophages in vivo are lacking. We designed a transgenic mouse model in which luminescence and fluorescence proteins, click beetle red luciferase (CBRED2) and mKate2, report on the expression of the Mrc1/Cd206 promoter, active in the monocyte/macrophage population. The mouse line was named B6Mrc1-mKate2-CBRED2. Using this novel mouse model, we were able to develop in vitro assays to validate transgenic macrophage polarization and test them with compounds of repolarization potency. Furthermore, in the in vivo assays, we exploited the migratory and infiltrative potency of macrophages for detecting tumor locations via optical imaging. In fact, macrophages can act as universal cancer markers, as they infiltrate primary and secondary tumors, stimulating or suppressing tumor growth. We first characterized transgenic mice for reporter expression ex vivo, followed by the generation of luminescence-based assays to reflect the polarity of differentiated macrophages, and lastly, we visualized reporter macrophages accumulating and infiltrating the tumor microenvironment (TME) of murine pancreatic ductal adenocarcinoma (PDAC) at multiple time points. We found that the extent of macrophage recruitment and retention was dependent on the infiltrative T-cell and dendritic cell populations present in the TME, reflecting the immunologically hot or cold nature of the PDAC clones, respectively. In conclusion, the ability to optically detect light-emitting macrophages can be applied not only for cancer studies but also in the context of inflammatory diseases. Full article

Cancer immunotherapy has transformed oncology by harnessing the immune system to recognize and eliminate malignant cells. However, many cancers exhibit limited or variable responses to this class of treatment due to insufficient antigen presentation and impaired interferon (IFN) signaling, creating an immunologically “cold” tumor microenvironment (TME) characterized by poor immune cell infiltration and treatment resistance. Viral mimicry has emerged as a therapeutic strategy to overcome these limitations by reactivating innate antiviral pathways within tumor cells. Viral mimicry occurs through the reactivation of endogenous retroviruses (ERVs) and other retrotransposons (e.g., LINE-1), which subsequently stimulate downstream nucleic acid sensing pathways. The resulting type I/III IFN responses restore antigen presentation and attract cytotoxic immune cells, sensitizing resistant tumors to immunotherapy. However, systemic stimulation of these pathways can trigger context-dependent inflammation and adaptive resistance, highlighting the need for temporal and spatial control. In this review, we examine the mechanistic foundation and clinical trajectory of viral mimicry, with an emphasis on its potential integration with established treatments and engineered immune cell platforms. By identifying the molecular and clinical gaps, viral mimicry can be harnessed to enhance tumor-specific immune activation and overcome treatment resistance in cancer immunotherapy. Full article

Background: Post-stroke upper-limb spasticity can cause pain, hinder passive care, and lead to secondary musculoskeletal complications. Current minimally invasive treatments have important limitations. Cryoneurolysis, which creates a controlled cold lesion of peripheral nerves, may offer a partially reversible focal denervation alternative. Methods: We conducted a feasibility case series in the outpatient department of a rehabilitation centre. Three adults with chronic post-stroke hemiparesis and a non-functional spastic upper limb underwent ultrasound- and nerve stimulation-guided cryoneurolysis of the musculocutaneous, median, and/or ulnar nerves. All had demonstrated a positive response to diagnostic nerve blocks beforehand. Feasibility outcomes included completion of planned nerve targets, tolerability under local anesthesia, absence of serious adverse events, and completion of 6-month follow-up. Secondary outcomes were Modified Ashworth Scale (MAS), qualitatively assessed passive joint mobility (video-documented), pain measured by visual analogue scale, sensory testing, and electroneuromyography (ENMG). Results: All procedures were completed as planned. Treatment was well tolerated under local anesthesia, and no serious adverse events occurred. MAS decreased by at least 2 points in targeted patterns, with immediate improvement in passive mobility; these effects persisted at 6 months. Pain remained unchanged in two participants and improved in one. Sensory testing at 6 weeks was stable. ENMG findings were heterogeneous, including reduced ulnar sensory action potential amplitude and biceps denervation activity in one participant. Conclusions: In this small series, cryoneurolysis for post-stroke upper-limb spasticity was feasible and associated with sustained tone reduction and improved passive mobility. Larger controlled studies are required to better define safety, optimize targeting strategies, and assess patient-centred outcomes. Full article

Cold exposure is an unavoidable stressor in cold regions, leading to growth retardation, oxidative damage, and endocrine disruption. This study investigated changes in blood parameters and antioxidant function in the brown adipose tissue (BAT) of mice exposed to cold. Sixteen naturally mated female mice (aged 70 days) were selected and divided into a control group (CON, n = 8, 25 ± 1 °C) and a cold exposure group (CE, n = 8, 4 ± 1 °C). Each pregnant female gave birth to approximately 12 pups, and the litter (dams and pups co-housed) served as the independent experimental unit, with both euthanized for sampling when the pups reached 20 days of age. Results showed that cold exposure increased ADFI and ADG but decreased the feed conversion rate (FCR) in lactating mice. It also decreased platelet count (PLT) and mean corpuscular hemoglobin concentration (MCHC), elevated lactate dehydrogenase (LDH) activity, and decreased TG and non-esterified fatty acid (NEFA) levels. Hormonal changes included increased adrenocorticotropic hormone (ACTH), apelin 12 (AP12), INS, NE, decreased cortisol (COR), LEP, and thyroid-stimulating hormone (TSH). In pups, cold exposure inhibited growth, reduced PLT, plateletcrit (PCT), red blood cells (RBC), and hemoglobin (HGB), altered lipid profiles, and induced hormonal shifts. Notably, cold exposure enhanced the BAT antioxidant capacity in pups, increasing the total antioxidant capacity (T-AOC) and antioxidant enzyme activities, as supported by gene expression. These findings suggest that, despite growth suppression, mice maintain homeostasis by modulating blood parameters and enhancing BAT antioxidant function to mitigate cold-induced damage. Full article

Adverse environmental and postharvest conditions challenge the functional quality of lemons, an economically vital citrus crop. Melatonin (MEL) has emerged as an effective regulator of plant stress responses and secondary metabolism. This study evaluated the effects of pre- and postharvest MEL treatments, combined with cold storage, on the fruit quality of two lemon cultivars (‘Fino’ and ‘Verna’). The research focused specifically on endogenous MEL and flavanone dynamics. Three experimental conditions were assessed: (a) preharvest MEL application at 0.1 and 1 mM; (b) preharvest treatment followed by cold storage; and (c) combined pre- and postharvest MEL treatment followed by cold storage. Preharvest treatments increased endogenous MEL at harvest in a dose- and cultivar-dependent manner. Specifically, 1 mM being optimal for ‘Fino’, while 0.1 mM was more effective for ‘Verna’. During cold storage, ‘Fino’ fruit, characterized by low basal endogenous MEL levels, showed a marked increase in MEL accumulation, suggesting the stimulation of biosynthesis. In contrast, ‘Verna’ fruit, which had initially high endogenous MEL content, exhibited a pronounced decline, indicating MEL consumption to counteract oxidative stress. Flavanone content increased dose-dependently after preharvest treatment and was preserved during storage in ‘Fino’ but declined in ‘Verna’. These findings demonstrate that the fruit cultivar must be considered a critical factor in MEL-based strategies, as identical treatments may yield markedly different outcomes even within the same species. Full article

Autophagy is a fundamental lysosome-dependent degradation process that maintains cellular homeostasis in response to stress. VSP34 (Vacuolar Protein Sorting 34, PIK3C3) is the only class-III phosphatidylinositol 3-kinase and generates phosphatidylinositol 3-phosphate (PI3P) for auto-phagosome nucleation and maturation. Thus, it provides a critical adaptive survival pathway for cells that are experiencing metabolic stress. The VPS34–autophagy axis plays dual roles in cancer, which depend on the context: it can restrain early tumorigenesis, but in established tumors, it can promote survival in conditions of hypoxia, nutrient deprivation, and therapeutic pressure. Moreover, VPS34 shapes the tumor microenvironment (TME) through its influence on both immune and cancer cells by modulating autophagy, cGAS-STING (cyclic GMP-AMP synthase Stimulator of Interferon Genes), and STAT1 pathways. VPS34 inhibition has been reported to induce an interferon response that increases CD8⁺ T and natural killer (NK) cell infiltration and converts cold tumors into hot ones. This behavior suggests that combining VPS34 inhibitors with cancer immunotherapies could be beneficial. In this review, we summarize the molecular functions and regulations of VPS34 in autophagy and discuss recent advances linking VPS34 to tumor and cancer immunotherapy. Full article

Gut ischemia/reperfusion (I/R) injury releases damage-associated molecular patterns (DAMPs), such as extracellular cold-inducible RNA-binding protein (eCIRP). Milk fat globule–epidermal growth factor VIII-derived oligopeptide 3 (MOP3) is a novel peptide enabling macrophage uptake of eCIRP via αvβ3-integrin. MOP3 reduces inflammation in gut I/R, but its mechanisms are not completely understood. We hypothesized MOP3 promotes macrophage polarization toward an anti-inflammatory, M2-like phenotype in gut I/R. We induced gut I/R in mice through 60 min of superior mesenteric artery occlusion followed by 4 h of reperfusion. Intestines were evaluated for macrophage polarization by flow cytometry and immunofluorescence histology. Peritoneal cavity macrophages were isolated from mice and treated with eCIRP, MOP3, α_vβ₃-antibody, and/or naïve IgG for 4 or 24 h. Polarity was assessed by flow cytometry, qPCR, and ELISA. Compared to the sham, the M2 proportion after gut I/R decreased by 22.7%, and the M1 proportion increased by 241%. MOP3 treatment increased the M2 proportion by 64.3%, and the M1 proportion decreased by 22.7%. In eCIRP-stimulated macrophages, MOP3 treatment increased M2-like and reduced M1-like cell-surface markers, gene expression, and cytokine levels. α_vβ₃ antibody dramatically reduced MOP3′s effects. MOP3 promotes M2 polarization through α_vβ₃ integrin-mediated clearance of eCIRP, a novel mechanism whereby MOP3 reduces gut I/R injury. Full article