Ovalbumin Research Articles

Hydrogen Gas Inhalation Alleviates Airway Inflammation and Oxidative Stress on Ovalbumin-Induced Asthmatic BALB/c Mouse Model

Airway inflammatory diseases, such as asthma, are a global public health concern owing to their chronic inflammatory effects on the respiratory mucosa. Molecular hydrogen (H2) has recently been recognized for its antioxidant and anti-inflammatory properties. In this study, we examined the therapeutic potential of H2 in airway inflammation using an ovalbumin (OVA)-induced BALB/c mouse model of allergic asthma. Female BALB/c mice were sensitized and challenged with OVA to induce airway inflammation, and 30 mice were randomly divided into five groups: NT (non-treatment), HTC (3% H2 treatment only), NC (negative control, OVA only), PC (positive control, OVA + intranasal 1 mg/mL salbutamol 50 μL), and HT (H2 treatment, OVA + inhaled 3% H2). Various inflammatory and oxidative stress (OS)-induced markers such as white blood cells (WBCs) and their differential counts, lung histology, cytokine levels such as interleukin (IL)-4, (IL)-5, (IL)-13, interferon-gamma (IFN-γ), tumor necrosis factor-alpha (TNF-α), granulocyte-macrophage colony-stimulating factor (GM-CSF), (IL)-10, reactive oxygen species (ROS), nitric oxide (NO), glutathione peroxidase (GPx), and catalase (CAT), and total immunoglobulin E (IgE) levels were investigated. Our results showed that inhaled H2 significantly reduced inflammatory cell infiltration, OS markers, and pro-inflammatory cytokine expression while upregulating antioxidant enzyme activity. Furthermore, H2 also significantly decreased serum IgE levels, a marker of allergic inflammation. Collectively, our findings suggest that H2 inhalation is a promising treatment option for airway inflammation, offering a novel approach with potential clinical applications.

P-type pilus PapG protein elicits toll-like receptor 2-mediated immune activation during cancer immunotherapy

The immune activation ability of FimH, an adhesion protein in pili of Escherichia coli (E. coli), has been recently reported. However, studies on the immune activity of PapG, another major pili terminal protein, have not been well explored. In this study, the immune stimulatory effect of purified recombinant PapG was evaluated. PapG treatment promoted dramatic changes in dendritic morphology of the bone marrow-derived dendritic cells (BMDCs) and induced upregulation of co-stimulatory molecule levels, major histocompatibility complex (MHC) I and II expression, and pro-inflammatory cytokine production in BMDCs. To identify the stimulatory receptor of PapG, an in silico study was performed. PapG exhibited strong binding affinity with murine toll-like receptor 2 (TLR2). In addition, PapG-induced activation of splenic DC and its subsets was unsuccessful in TLR2-knock out mice. Combination of PapG and ovalbumin (OVA) elicited OVA-specific T cell proliferation and cytokine production and cytotoxicity that consequently promoted anti-cancer immune responses against OVA-expressing B16 melanoma. Furthermore, PapG treatment induced activation of peripheral blood DCs and its subsets in humans in a TLR2 dependent manner. PapG-stimulated human conventional DC2 promoted syngeneic T cell proliferation and activation. The findings of this study demonstrated that PapG could be a useful immune stimulator for immunotherapy against cancer.

Quantitative Analysis, Anti-Inflammatory and Analgesic Effects of Ethanol Leaf Extract and Fractions of Microsorium scolopendria (Burm. f.) Copel. in Mice

Microsorium scolopendria, a fern was studied for phytochemicals, anti-inflammatory, and analgesic effects to add credence to its various folkloric applications. Phytochemicals were analyzed quantitatively, anti-inflammatory experiment was carried out with egg albumin- induced paw oedema and xylene-induced ear oedema models while analgesic effects were studied using formalin-induced paw licking, acetic acid-induced writhing and hot plate-induced pain models. The results revealed the presence of alkaloids, flavonoids, saponins, tannins, terpenoids, and phenols with alkaloids (14.6 %w/w) as the most predominant phytoconstituent. In anti-inflammatory experiments, M. scolopendria extract reduced oedema caused by egg albumin and xylene in a dose related manner and comparable to standard agents. Also, in analgesic experiments, the extract reduced pain induced by formalin, acetic acid, and hot plate. These reductions were statistically (p?0.05) significant. Considering the fractions in all experiments, butanol and ethyl acetate fractions were prominent in anti-inflammatory effect while ethyl acetate fraction was at top in reduction of pain. This study supports the use of M. scolopendria in ethnomedicinal practice.

Targeting TMEM16A ion channels suppresses airway hyperreactivity, inflammation, and remodeling in an experimental Guinea pig asthma model

Asthma is a chronic inflammatory disease characterized by airway hyperresponsiveness, inflammation, and remodeling. Calcium (Ca2+)-activated chloride (Cl−) channels, such as TMEM16A, are inferred to be involved in asthma. Therefore, the present study investigated the therapeutic potential of TMEM16A inhibition in a guinea pig model of ovalbumin (OVA)-induced allergic asthma. Guinea pigs were treated with a specific blocker, CaCCinh-A01 (10 μM), administered via inhalation. A significant reduction in cough reflex sensitivity and specific airway resistance was observed in animals treated with CaCCinh-A01, highlighting its potential to improve airway function. Despite a reduction in ciliary beating frequency (CBF), CaCCinh-A01 reduced airway mucus viscosity by decreasing the production of mucin-5AC (MUC5AC). The nonspecific reduction in the Th1/Th2 cytokine spectrum following CaCCinh-A01 treatment indicated the suppression of airway inflammation. Additionally, markers associated with airway remodeling were diminished, suggesting that CaCCinh-A01 may counteract structural changes in airway tissues. Therefore, inhibition appears to mitigate the pathological aspects of asthma, including airway hyperresponsiveness, inflammation, and remodeling. However, further studies are required to comprehensively evaluate the potential of TMEM16A as a therapeutic target for asthma.

Unravelling The Bioactivities of Acmella paniculata Extract-Mediated Green Deep Eutectic Solvent of Citric Acid Monohydrate and Glycerol

Plants are important sources of underlying medicinal value properties. The extraction of bioactive compounds from botanical sources using green solvents has gained interest due to its environmental sustainability. This study highlighted the bioactivities potential of Acmella paniculata extract mediated by green deep eutectic solvent (DES) composed of the citric acid monohydrate and glycerol. Acmella paniculata, a local flowering shrub was selected due to its rich medicinal value compounds. The synergistic effect between plant’s bioactive compounds and DES is capable of enhancing bioactivity, making DES a promising plant solvent extractor candidate. The plant extracts were prepared in leaf and flower parts using the centrifugation method. The phytochemical screening for both extracts showed the presence of terpenoids and steroid constituents which have valuable bioactivity functions. The antibacterial activity assessed by disc diffusion assay exhibited higher susceptible bacterial response of E. coli, Salmonella enterica ser. Typhimurium and S. aureus against the flower extract compared to the leaf extract. The DPPH assay was conducted to assess free radical scavenging activity. The flower extract demonstrated radical scavenging activity (RSA) of 75%-77% while the leaf extract demonstrated 65%-69%. The flower extract results showed higher RSA emphasizing its potential as a natural antioxidant. The anti-inflammatory activity was determined by egg albumin denaturation assay, which showed a greater inhibition rate in flower extract than the leaf extract which was up to 95% and 89% respectively. Thus, both extracts possess an in vitro anti-inflammatory effect. Conclusively, flower extract exhibited better bioactivities value than leaf extract in a green DES. Hence, offering a new insight into its application as an effective alternative in natural product-based therapeutics.

In-situ synthesis of 2D nanozymes-coated cellulose nanofibers on paper-based chips for portable detection of biothiols

BackgroundSimple, fast and low-cost paper-based analytical devices (PADs) have a good application prospect for point-of-care detection of GSH. However, effective immobilization of functional nanomaterials onto cellulose, as a critical factor in the construction of PADs, presents numerous difficulties and challenges. ResultsIn this study, we have developed an exceptionally straightforward and environmentally friendly synthetic approach by using ovalbumin (OVA) as a bio-mineralization template for the preparation of MnO2 nanosheets. The MnO2 nanosheets produced in the solution phase exhibited excellent intrinsic nano-enzyme activity and biodegradability. The OVA-MnO2 nanosheets can effectively oxidize Amplex red in the absence of H2O2, enabling sensitive detection of GSH with a linear range of 5 nM–10 μM and a detection limit as low as 2.8 nM. Furthermore, we utilized this method to facilitate in situ synthesis of OVA-MnO2 nanosheets directly on paper substrates. This approach eliminates the need for conventional stirring and centrifugation steps, greatly simplifying the fabrication process while reducing material usage and time expenditure. Characterization of the chemical composition and morphology confirmed the intimate growth of the 2D nano-enzymes on the cellulose fibers. Utilizing smartphone capabilities, the OVA-MnO2 nanosheet-modified PAD enabled instrument-free detection of GSH, demonstrating high sensitivity (0.74 μM) and a wide linear response range (1–1000 μM). SignificanceThe synthesis of MnO2 nanosheets directly on cellulose substrates substantially streamlines the modification workflow of PADs and reduces detection costs, offering new avenues for clinical diagnostics of relevant diseases.

Electroacupuncture preconditioning prevents urticaria by regulating inflammatory response in rats with urticaria

To observe the effect of electroacupuncture (EA) preconditioning at "Quchi" (LI11) and "Xuehai" (SP10) in prevention of urticaria. Twenty-four male SD rats were randomly divided into control, model and preconditioning of EA (Pre-EA) groups (8 rats/group). The urticaria model was established by intradermal injection of dilute allogeneic antioalbumin serum at the spots of the bilateral symmetry of the spine on the back, and followed by tail venous injection of mixture solution of egg albumin diluent, plus 0.5% Evans blue and normal saline. Ten days before the end of modeling, rats of the pre-EA group received EA stimulation of LI11 and SP10 for 20 min, once a day for 10 consecutive days. The times of rat's scratching the sensitized skin were recorded. HE staining method was used to observe the pathological changes of skin tissue, and toluidine blue staining method was used to observe the morphology of mast cells (MCs) in the skin, blood, mesentery, and peritoneal fluid, and calculate the degranulation rate. Immunohistochemical stainning was used to detect immunoglobulin E (IgE), histamine (HIS), and 5-hydroxytryptamine (5-HT) expressions in subcutaneous tissue. NOD like receptor thermal domain associated protein 3 (NLRP3) inflammasome, apoptosis related granule protein (ASC), and cysteine aspartate aminotransferase 1 (Caspase-1) protein expression levels in skin tissue were detected by Western blot. The contents of serum interleukin(IL)-1β and IL-18 were detected using ELISA method. Compared with the control group, the scratching times, amount of Evans blue exudation of the sensitized blue spots, degranulation rate of MCs in skin, blood, mesentery and peritoneal fluid, the expression levels of IgE, HIS, 5-HT in subcutaneous tissue, protein expression levels of NLRP3, ASC, Caspase-1 in skin tissue, and the contents of serum IL-1β and IL-18 were significantly increased (P<0.01, P<0.05) in the model group. In comparison with the model group, the scratching times, amount of Evans blue exudation of the sensitized blue spots, degranulation rate of MCs, the expression levels of IgE, HIS, 5-HT in subcutaneous tissue, protein expression levels of NLRP3, ASC, Caspase-1 in skin tissue, and the contents of serum IL-1β and IL-18 in EA group were significantly decreased (P<0.01, P<0.05). EA preconditioning at LI11 and SP10 can prevent and treat UR by inhibiting inflammatory response, which is related to the regulation of pyroptosis.

Abietic acid antagonizes the anti-inflammatory effects of celecoxib and ketoprofen: Preclinical assessment and molecular dynamic simulations

The present work is designed to explore the anti-inflammatory properties of AA and its modulatory effects on celecoxib (CEL) and ketoprofen (KET) through in vitro, ex vivo, in vivo, and in silico approaches. Different concentrations of AA were utilized to evaluate the membrane-stabilizing potential via egg albumin and the Human Red Blood Cell (HRBC) denaturation model. In the animal model, formalin (50 μL) was injected into the right hind paw of young chicks to induce inflammation. AA was administered at 20 and 40 mg/kg (p.o.) to the experimental animals. We used CEL and KET as positive controls. The vehicle was provided as a control group. Two combinations of AA with CEL and KET were also investigated in all tests to assess the modulatory activity of AA. In addition, in silico investigation was used for predictions about drug-likeness, pharmacokinetics, and toxicity of the selected chemical compounds, and the study also evaluated the binding affinity, visualization, and stability of ligand-receptor interactions through molecular dynamic (MD) simulation. Results manifested that AA concentration-dependently significantly inhibited the egg albumin denaturation (IC50: 27.53 ± 0.88 μg/ml) and breakdown of HRBC (IC50: 15.69 ± 0.75 μg/ml), indicating the membrane stabilizing potential compared to the control group. AA also significantly (p < 0.05) lessened the frequency of licking and alleviated the paw edema in a dose-dependent manner in an in vivo test. However, AA reduced the activity of CEL and KET in combination treatment. AA showed good pharmacokinetic characteristics to be considered as a therapeutic candidate. Additionally, the in silico study displayed that AA demonstrated a relatively higher docking score of −9.1 kcal/mol with the cyclooxygenase-2 (COX-2) enzyme and stable binding in MD simulation. Whereas the standard ligand (CEL) expressed the highest binding value of −9.2 kcal/mol to the COX-2.

Copper Oxide Nanoparticles Induce Pulmonary Inflammation and Exacerbate Asthma via the TXNIP Signaling Pathway.

Copper oxide nanoparticles (CuO NPs) have seen increasing use across various industries, raising significant concerns about their potential toxicity and the exacerbation of pre-existing conditions like asthma. Asthma, a chronic inflammatory condition of the airways, can be triggered or worsened by environmental factors such as allergens, air pollutants, and chemicals, including nanoparticles. This study aimed to investigate the pulmonary toxicity induced by CuO NPs and their impact on asthma, with a particular focus on the role of thioredoxin-interacting protein (TXNIP). Using an ovalbumin (OVA)-induced asthma model, we found that CuO NP exposure led to significant increases in inflammatory cell infiltration, cytokine production, airway hyperresponsiveness, OVA-specific immunoglobulin (Ig)E levels, and mucus production. These pathological changes were closely associated with the upregulation of TXNIP-related signaling pathways, including phosphorylated apoptosis signal-regulating kinase (p-ASK)1, the Bax/Bcl-2 ratio, and cleaved caspase-3 activation. Complementary in vitro experiments using NCI-H292 respiratory epithelial cells showed that CuO NP treatment enhanced TXNIP signaling and increased mRNA expression and the production of inflammatory cytokines. Notably, TXNIP knockdown significantly attenuated these CuO NP-induced effects. In conclusion, our findings suggest that CuO NP exposure not only induces pulmonary toxicity but also exacerbates asthma, primarily through the activation of the TXNIP signaling pathway.

Enhanced immunomodulatory properties of ovalbumin through transglutaminase and tyrosinase/caffeic acid-catalyzed crosslinking plus galactomannan conjugation alleviates allergic asthma

Ovalbumin (OVA) is the primary allergenic protein, associated with T helper 2 (Th2)-mediated allergy reactions. Here, we studied OVA’s conformational structure, digestibility, and allergenic potency upon galactomannan (Man) conjugation along with transglutaminase-catalyzed crosslinking (TG-Man/OVA) and tyrosinase/caffeic acid-catalyzed crosslinking (Tyr-Man/OVA). Enzymatic glycosylation altered OVA’s conformation and enhanced the gastrointestinal digestibility. Stimulation of bone marrow-derived dendritic cells (BMDCs) with enzymatically glycosylated OVA reduced the expression of CD40, CD80, CD86 and MHC class II molecules. Furthermore, glycosylated OVA increased IL-10 production while suppressing proinflammatory cytokine production in BMDCs. In an OVA-induced mouse asthma model, TG-Man/OVA and Tyr-Man/OVA upregulated IFN-γ and IL-10 expression and downregulated IL-4, IL-5, and IL-13 in lungs. Crosslinked OVA diminished infiltrated cells in bronchoalveolar lavage fluid and lung and attenuated eosinophilic airway inflammation. These findings offer valuable insights into the development of a novel product with improved hypoallergenic and immunomodulatory properties and hold promise in attenuating allergic diseases.

Effect of Nicotinamide Riboside Against the Exhaustion of CD8+ T Cells via Alleviating Mitochondrial Dysfunction.

Background: Targeting mitochondria and protecting the mitochondrial function of CD8+ T cells are crucial for enhancing the clinical efficacy of cancer immunotherapy. Objectives: In this study, our objective was to investigate the potential of nicotinamide riboside (NR) in preserving the mitochondrial function of CD8+ T cells and mitigating their exhaustion. Methods: We established two in vitro models to induce CD8+ T cell exhaustion either by tumor cell-conditioned medium (TCM) or by continuous stimulation with OVA(257-264) peptide. CD8+ T cells were treated in the absence/presence of NR. Results: Our findings demonstrated that NR supplementation effectively inhibited CD8+ T cell exhaustion and preserved mitochondrial function in both models. Moreover, apoptosis of CD8+ T cells was reduced after NR treatment. Western blot data indicated that NR treatment upregulated Silent information regulator 1 (SirT1) expression. Further inhibition of Sirt1 activity using EX527 uncovered that the inhibitory effect of NR on CD8+ T cell exhaustion and its protective effect on mitochondria were attenuated. Conclusions: In conclusion, our results indicate that NR supplementation attenuates CD8+ T cell exhaustion, and its underlying mechanism is associated with increased mitochondrial function regulated by the SirT1 pathway. Our research provides evidence that NR may assist in enhancing the clinical efficacy of immunotherapy.

Shikonin alleviates asthma phenotypes in mice via an airway epithelial STAT3-dependent mechanism.

Asthma is an inflammatory disease where the balance between Th1/Th2 and Th17/Treg plays a crucial role in its pathogenesis. Shikonin is used to treat a variety of autoimmune diseases due to its good anti-inflammatory activity. However, the effect and mechanism of shikonin on asthma remain unknown. Mice were sensitized with ovalbumin (OVA)/house dust mite (HDM) and treated with shikonin. Lung inflammation was assessed histologically and via flow cytometry. Bronchoalveolar lavage fluid (BALF) was analyzed for cell counts and cytokines. Shikonin's impact on p-STAT3 was studied in vivo and in vitro. Shikonin inhibited OVA or HDM-induced inflammation and airway hyperresponsiveness. Upon treatment, a restoration of the Th1/Th2 and Th17/Treg balance was observed, evidenced by a reduction in IL-4 and IL-17A levels in BALF, alongside an elevation in interferon-gamma and IL-10. Furthermore, shikonin impeded the infiltration of eosinophils, neutrophils, macrophages, and lymphocytes into lung tissue. The observed decrease in STAT3 phosphorylation and diminished nuclear translocation of p-STAT3 confirmed that shikonin promotes the balance of Th1/Th2 and Th17/Treg by regulating airway epithelial STAT3. Shikonin mitigates asthma symptoms through a STAT3-dependent mechanism, indicating its potential as an anti-asthmatic therapeutic agent.

Conjugation of Multiple Proteins Onto the Surface of PLGA/Lipid Hybrid Nanoparticles.

Nanoparticles are increasingly being used in the development of vaccines for disease prevention or treatment. Recent research has demonstrated that conjugating a protein onto the surface of nanoparticles can significantly increase its immunogenicity. Considering various pathogens that threaten human health, multivalent vaccines are often desirable. Up to now, nanoparticle-based vaccines are mostly limited to one protein per nanoparticle. No research has been conducted to explore the possibility of conjugating more than one protein onto the surface of a nanoparticle. Here we developed a specific conjugation strategy to conjugate multiple proteins to the PLGA/lipid hybrid nanoparticle surface. The maleimide-thiol Michael addition, Aizde-DBCO (Dibenzocyclooctyne), and TCO (trans-cycloctene)-Tetrazine click chemistry were employed to conjugate three different proteins, subunit keyhole limpet hemocyanin (sKLH), Ovalbumin (OVA), and cross-reactive material 197 (CRM197), to the surface of PLGA/lipid hybrid nanoparticles (hNPs). The successful results of this study pave the way for developing multivalent vaccines against different pathogens.

Enhancing preventive and therapeutic cancer vaccine efficacy through biotherapeutic ligand-associated extracellular vesicles

Extracellular vesicles (EVs), secreted by almost all living cells, have gained significant attention for their role in intercellular communication and their potential as versatile carriers for biotherapeutics. However, the clinical translation of EV-based therapies faces significant challenges, primarily due to the lack of efficient methods for loading biotherapeutic agents into EVs. This study introduces a simple, reproducible strategy for the simultaneous incorporation of various biotherapeutics within EVs. The process is gentle and preserves the essential physicochemical and biological characteristics of EVs, thereby protecting labile ligands from premature degradation and elimination. The binding and uptake efficiency of EVs by target cells reached approximately 97 % within 24 h of incubation. Administration of EVs loaded with oligodeoxynucleotides (ODN) resulted in a 4-fold increase in IFNγ+ CD4+ T cells and a 5-fold increase in IFNγ+ CD8+ T cells in the spleens and lymph nodes. Additionally, the co-administration of EVs with ODN and ovalbumin (OVA) induced elevated Th1-biased antibody responses and antigen-specific cytotoxic T-cell responses, providing long-lasting complete protection in 60 % of mice against T-cell thymoma challenge. Furthermore, EVs associated with three different ligands (OVA, CpG-ODN, and α-GalCer) effectively regressed established murine melanoma and significantly improved survival rates in mice. This study presents a powerful and promising approach to overcoming the limitations of EV-based cancer vaccines, advancing the development of effective cancer immunotherapies. SummaryImmunization with EVs that are co-associated with antigen and biotherapeutic cargo through a lyophilization-based technique elicits potent anti-cancer immunity.

Antigenicity elimination of ovalbumin by cold plasma-induced covalent binding with Gallic acid

The effect of cold plasma (CP) treatment in promoting the covalent grafting of ovalbumin (OVA) with gallic acid (GA) were investigated, along with identifying the binding sites in the OVA-GA complex and exploring its potential for reducing the antigenicity of OVA. The results showed that the GA content of 22.97 ± 1.27 mg/g in OVA-GA complex was obtained following 60 s of CP treatment. Using LC-MS/MS, four regions (T52-R59, V201-K207, I279-R285, and V281-K291) were identified, containing 12 GA binding sites in the OVA-GA complex. Additionally, a significant reduction in IgE binding capacity (70.83 ± 0.90 %) was observed, as confirmed by ELISA analysis. The masking/steric-hindrance effect on linear epitopes and the disruption of conformational epitopes of OVA as a result of GA grafting may be the key factors leading to the reduction in OVA antigenicity. This study highlights that promoting the grafting of polyphenols onto proteins using CP treatment is an effective strategy for reducing the antigenicity of protein allergens.

A Mixture of Four Probiotic Strains (Probiatop®) Mitigates Food Allergy to Ovalbumin in Mice.

Food allergy is a pathological condition caused by a disruption of oral tolerance. This condition leads to a pro-inflammatory environment that culminates in symptoms that varies from abdominal pain to anaphylaxis and death. The gold standard treatment consists of removing the food that triggers the allergy from diet. However, this conduct can cause nutritional impairment and social restrictions. Therefore, the need for new treatment strategies is notorious. In this context, probiotics are investigated due to their immunomodulatory mechanisms. Therefore, the objective of the present work is to investigate the probiotic potential of a mixture of four probiotic strains (Probiatop®) in an in vivo model of food allergy to ovalbumin (OVA). Our results demonstrated that oral administration of Probiatop® attenuated weight loss and diminished significantly anti-OVA IgE and IgG1 levels. Furthermore, it mitigated proximal jejunum injury, neutrophil recruitment and local IL17 levels. In addition, the probiotic mixture modulated positively the gut microbiota composition by decreasing the levels and frequency of Staphylococcus and yeast. In summary, our data suggest that Probiatop® has the potential to alleviate important symptoms of IgE-mediated food allergy, suggesting its probiotic potential as an adjuvant in the treatment of ovalbumin food allergy.

GBP2 Regulates Lipid Metabolism by Inhibiting the HIF-1 Pathway to Alleviate the Progression of Allergic Rhinitis.

Allergic rhinitis (AR) is a prevalent allergic disorder instigated by a variety of allergenic stimuli. The study aims to elucidate the mechanistic underpinnings of Guanylate-binding protein 2 (GBP2) in modulating AR. Bioinformatics analysis was used to identify hub genes in AR, and GBP2 was identified. Mice were injected with ovalbumin (OVA) to create AR model. The pathological changes of the nasal mucosa were observed by hematoxylin-eosin staining. ELISA and western blot demonstrated that in OVA-induced AR mice, high IgE and IgG1 levels, inflammation (increased TNF-α, IL-5 and IFN-γ), oxidative stress (high ROS, low TAOC and GSH) and abnormal lipid metabolism (increased TC and LDL-C, decreased HLD-C) were observed. Mouse nasal mucosal epithelial cells (MNECs) were treated with TNF-α to simulate AR. Cell viability and apoptosis were evaluated by CCK-8 assay and flow cytometer, respectively. In vitro assay revealed that GBP2 inhibited total IgE, OVA-IgE and IgG1 levels and suppressed abnormal lipid metabolism, inflammation and oxidative stress to alleviate AR. Furthermore, HIF-1 pathway was screened as the downstream pathway of GBP2. GBP2 inhibited the HIF-1 pathway, and Fenbendazole-d3, the activator of HIF-1 pathway, weakened the inhibitory effects of GBP2 on apoptosis, inflammation, oxidative stress and abnormal lipid metabolism in vitro. In summary, GBP2 alleviated abnormal lipid metabolism, inflammation and oxidative stress by inhibiting the HIF-1 pathway, providing a direction for the treatment of AR.

Comparison of Essential Oils of Schinus molle L. Leaves Growing in Different Regions of Algeria in Terms of Their Chemical Compositions and Various Biological Activities

ABSTRACTThis preliminary study investigated the chemical composition and biological properties of essential oils derived from Schinus molle L. leaves obtained from three specific regions in Algeria: Algiers (SMA), Djelfa (SMD), and Mascara (SMM). Gas chromatography (GC) and Gas chromatography–Mass Spectrometry (GC–MS) analyses confirmed the presence of 52 compounds (100.0%) in SMA essential oil, 50 compounds (100.0%) in SMD, and 72 compounds (100.0%) in SMM essential oils. The main components of SMA oil were camphene (31.82%), limonene (14.71%), and p‐cymene (9.25%). In SMD and SMM oils, α‐phellandrene (14.25% and 12.70%), limonene (13.02% and 11.90%), and germacrene D (10.62% and 10.15%) were the major components, respectively. The antioxidant characteristics were evaluated using five methods: β‐carotene‐linoleic acid, DPPH, ABTS+, CUPRAC, and Metal chelating assays. The results revealed a moderate to low anti‐oxidant effect, with SMA essential oils exhibiting the highest activity. A moderate inhibitory effect against acetylcholinesterase (AChE), butyrylcholinesterase (BChE), α‐glucosidase, α‐amylase, urease, and tyrosinase was observed, indicating anti‐enzymatic activity. Nevertheless, all samples had higher IC50 values for both antioxidant and anti‐enzymatic activities than 200 μg/mL. It was determined that regional differences in the locations where S. molle grows showed both qualitative and quantitative differences in essential oil components. This difference was also detected in the biological activities of the essential oils. The anti‐inflammatory capacities of the three samples were assessed using Human Red Blood Cell (HRBC) membrane stabilisation and egg albumin denaturation techniques. The results showed effective anti‐inflammatory activity in all three samples. The antimicrobial efficacy was evaluated using a range of tests, such as anti‐quorum sensing, violacein inhibition, anti‐swimming, and anti‐swarming assays, demonstrating moderate activity. No toxic effects were observed at the tested dosages when assessing cytotoxicity against a healthy cell line (CCD18‐Co). This thorough examination provides valuable insights into the chemical composition and bioactive properties of essential oils derived from S. molle. These findings indicate the possible use of these essential oils in the food, medicinal, and cosmetic industries.

Thermo- and pH-Responsible Gels for Efficient Protein Adsorption and Desorption

Protein adsorption behavior was examined on poly(N-isopropylacrylamide-co-sodium methacrylate)-based hydrogels at different temperatures: 5, 20, and 37 °C, and pH: 4.5, 7, and 9.2. The hydrogels, whose covalent skeleton contains pendant anionic units due to the presence of the sodium methacrylate co-monomer, exhibited both thermo- and pH-sensitivity with different extents, which depended on the content of ionizable moieties and the cross-linker density. The hydrogel composition, temperature, and pH influenced the zeta potential of the hydrogels and their swelling properties. The proteins selected for the study, i.e., bovine serum albumin (BSA), ovalbumin (OVA), lysozyme (LYZ), and a monoclonal antibody (mAb2), differed in their aminoacidic composition and conformation, thus in isoelectric point, molecular weight, electrostatic charge, and hydrophobicity. Therefore, the response of their adsorption behavior to changes in the solution properties and the hydrogel composition was different. LYZ exhibited the strongest adsorption of all proteins with a maximum at pH 7 (189.5 mg ggel−1); adsorption of BSA and OVA reached maximum at pH 4.5 (24.4 and 23.5 mg ggel−1), whereas mAb2 was strongly adsorbed at 9.2 (21.7 mg ggel−1). This indicated the possibility of using the hydrogels for pH-mediated separation of proteins differing in charge under mild conditions in a water-rich environment of both the liquid solution and the adsorbed phase. The adsorption affinity of all proteins increased with temperature, which was attributed to the synergistic effects of attractive electrostatic and hydrophobic interactions. That effect was particularly marked for mAb2, for which the temperature change from 5 to 37 °C caused a twentyfold increase in adsorption. In all cases, the proteins could be released from the hydrogel surface by a reduction in temperature, an increase in pH, or a combination of both. This allows for the elimination of the use of salt solution as a desorbing agent, whose presence renders the recycling of buffering solutions difficult.

The protective effects and mechanisms of essential oil from Chimonanthus nitens Oliv. leaves in allergic rhinitis based on the NF-κB and T-bet/GATA-3 signaling pathways

Ethnopharmacological relevancePreliminary studies showed that Shanlameiye granules are derived from Chimonanthus nitens Oliv. Leaves ameliorate inflammatory responses in mice with Allergic Rhinitis (AR). The essential oil from Chimonanthus nitens Oliv. Leaves (CLO) have been identified as the key active substances in these granules. However, whether CLO constitutes the primary mechanism for the mitigation of AR-related inflammation by these granules has not yet been investigated. Aim of the studyThis experiment was to validate the effects and mechanism of CLO on inflammatory responses in RAW264.7 cells and AR rat model. Materials and methodsAn inflammatory model was induced in RAW264.7 cells by Lipopolysaccharide (LPS) & Interferon-gamma (IFN-γ) stimulation. AR rat model was established using both systemic and local challenges with Ovalbumin (OVA). ResultsIn cell experiments, CLO obviously decreased the secretion of cytokines and inhibited the NF-κB signaling pathway activation. In animal experiments, CLO decreased the number of eosinophils in the blood and lowered the levels of cytokines in nasal lavage fluid (NALF). Additionally, CLO inhibited the expression of STAT6, GATA-3, and p-p65, while increasing the expression of STAT4 and T-bet in the nasal mucosa. ConclusionIn AR rat model, CLO may play an anti-inflammatory role in AR rat model by regulating NF-κB and T-bet/GATA-3 signaling pathways.