Search Results (52)

As the human population continues to grow, the demand for pork increases, and the management of infectious diseases in swine from a One Health standpoint is becoming more important than ever. To prevent antimicrobial use as much as possible, the search continues for alternative substances that can aid in mitigating oxidative stress and inflammation, which are cornerstones of infectious disease. In this study, we stimulated porcine peripheral mononuclear blood cells (pPBMCs) with either bacterial lipopolysaccharides (LPS) of different origin (Salmonella Typhimurium, Salmonella Enteritidis and E. coli), or the plant lectins concanavalin A (ConA) and phytohemagglutinin (PHA) to create an in vitro inflammatory model. Quercetin, a flavonoid with well documented positive effects, was used with the aim of decreasing oxidative stress and the production of the inflammatory cytokines IL-6 and IL-8. Oxidative stress was successfully induced in the pPBMCs by all stimulants (except for S. Enteritidis LPS), along with IL-6 production (except for E. coli LPS); IL-8 production was only induced by treatment with LPS. While quercetin had an antioxidant effect on the pPBMCs, it did not reduce IL-6 or IL-8 levels under the conditions tested and even had a pro-inflammatory effect by increasing IL-8 production when combined with LPS. To gain a deeper understanding of the immunomodulatory effects of quercetin on pPBMCs, further studies should be conducted to measure the production of additional pro- and anti-inflammatory cytokines, including TNF-α, IL-10, and IL-1β. Full article

Background/Objectives: Immune-mediated hepatitis, including autoimmune hepatitis, remains a formidable clinical challenge characterized by the rapid destruction of the liver parenchyma. While whey proteins are well-regarded for their anti-inflammatory properties, goat whey possesses a distinct bioactive profile, offering superior digestibility and reduced allergenicity compared to their bovine counterparts. This study investigated the hepatoprotective potential and underlying immunological mechanisms of lyophilized goat whey (LGW) in a Concanavalin A (ConA)-induced model of acute hepatitis. Methods: BALB/c and C57BL/6 mice were administered LGW orally (1 g/kg/day) for five consecutive days prior to a ConA challenge. Liver injury was quantified via serum transaminase levels and histopathological evaluation. The cytokine profiles and the phenotype of liver mononuclear cells (MNCs) were analyzed using ELISA and flow cytometry, respectively. Results: LGW pretreatment significantly attenuated ConA-induced hepatitis in both mouse strains, markedly reducing serum transaminase levels and preserving hepatic architecture. Mechanistically, LGW triggered a fundamental shift in the hepatic immune microenvironment by suppressing the pro-inflammatory Th1/Th17 axis (evidenced by decreased IFN-γ and IL-17) while concurrently upregulating the anti-inflammatory cytokine IL-10. Furthermore, LGW induced a tolerogenic phenotype in hepatic dendritic cells (CD11c⁺CD206⁺), which directly correlated with a significant expansion of regulatory T cells (Tregs). This strain-independent protection suggests that LGW modulates fundamental, early-stage immune signaling pathways within the liver. Conclusions: Our findings demonstrate that LGW exerts potent hepatoprotection by effectively reprogramming the hepatic immune microenvironment toward a tolerogenic state. These results position LGW as a promising, safe, and effective functional food candidate for the prevention and adjunct management of immune-mediated inflammatory liver diseases. Full article

Glycosylation plays a critical role in maintaining cardiac structure and function, yet its modulation during aging and hypertrophic cardiomyopathy (HCM) in feline hearts remains uncharacterized. This study provides a systematic analysis of lectin-binding patterns in feline myocardium across different age groups and disease states. Post-mortem feline hearts (n = 64), classified by age (newborn to senior) and diagnostic status (healthy vs. HCM-affected), were evaluated using tissue microarrays stained with five plant-derived lectins—Concanavalin A (ConA), Wheat Germ Agglutinin (WGA), RCA (Ricinus communis Agglutinin I), Tomato (Lycopersicon esculentum Agglutinin), and Griffonia (Bandeiraea) simplicifolia Lectin I (BS)—alongside Draq5 nuclear counterstaining. Lectin histochemistry revealed distinct, region-specific glycosylation patterns, with notable remodelling in both aged and HCM-affected hearts. These glycan alterations reflect underlying molecular and structural changes associated with cardiac aging and pathology. Although lectin histochemistry has been used to examine cardiac glycosylation in species such as mice, rats, zebrafish, and humans, comparable data for felines have been lacking, even if domestic cat represents a spontaneous model for human HCM. This study provides the first essential step in characterizing the feline cardiac glycosylation. The observed shifts in lectin-binding profiles reveal specific remodelling associated with aging and HCM in cats. These results provide a foundation for future studies assessing the utility of glycan motifs as potential post-mortem markers of disease progression in felines. Full article

This presented study depicts the synthesis of three novel carbazole–thiazole conjugates, thoroughly investigating their spectroscopic properties as well as evaluating their biological activity as tyrosinase inhibitors. Additionally, we investigated the possibility of using Concanavalin A (ConA) complexes with dyes from a theoretical point of view, developing a promising protein-based strategy of delivery of dyes to the target cells. The tyrosinase inhibition assay showed that compounds K1 and K3 demonstrated higher activity than the kojic acid with IC₅₀ values of 46 and 59 mM, respectively. Among the tested compounds, carbazole K3 exhibits the most pronounced nonlinear optical response due to its high electronic flexibility, strong solvatochromism, large excited-state dipole moments, and efficient intramolecular charge transfer. Additionally, all investigated carbazoles demonstrate high ability to form stable supramolecular complexes with ConA, which was confirmed using molecular docking studies. It was found experimentally and theoretically that the compound K3 has the best biophysical parameters, making it a promising candidate for potential diagnostic applications. Full article

Due to rising antibiotic resistance, it is necessary to develop alternative ways to combat pathogenic bacteria. One alternative is photodynamic antibacterial chemotherapy (PACT). This work presents the conjugation of the photosensitizer Rose Bengal (RB) to lectins to improve its efficacy against Gram-positive and Gram-negative bacteria. Two lectins, concanavalin A (ConA) and wheat germ agglutinin (WGA), were covalently linked to RB. Spectroscopic and chromatographic data confirmed successful conjugation. Microscopic examination demonstrated that both lectins agglutinate cells of Gram-positive S. aureus, including clinical multidrug-resistant MRSA strains, and Gram-negative E. coli, P. aeruginosa, and S. paratyphi B, although ConA showed a more pronounced reaction. Photodynamic assays showed that ConA-RB achieved complete eradication of S. aureus at significantly lower concentrations and light doses than free RB or WGA-RB. ConA-RB also exhibited higher efficacy against Gram-negative bacteria compared to free RB at lower concentrations and shorter illumination periods. WGA-RB was less effective, showing preferential activity against S. aureus. Our findings suggest that lectin–RB conjugates offer a promising approach for selective antibacterial treatment under illumination. Full article

Background: Ten new sesquiterpenes, including eight eremophilane-type sesquiterpenes (1–8) and two compounds (9–10) with a cyclopentane ring, representing an undescribed subtype of sesquiterpene, along with a new abietane-type diterpenoid (11), were isolated and identified from a deep-sea-derived fungus: Eutypella sp. Methods: Their structures were elucidated on the basis of various spectroscopic analyses, mainly including nuclear magnetic resonance (NMR) and high-resolution electrospray ionization mass spectrometry (HRESIMS) data, ¹³C NMR calculations with DP4+ probability analyses, electronic circular dichroism (ECD) calculations, and single-crystal X-ray diffraction experiments. Results: Furthermore, compound 11 exhibited potent immunosuppressive activity with IC₅₀ values of 8.99 ± 1.08 μM in a lipopolysaccharide (LPS) model and 5.39 ± 0.20 μM in a concanavalin A (ConA) model. Full article

The accurate detection and quantification of pathogenic bacteria is crucial for ensuring public health. In this work, we propose a sensitive and selective sandwich electrochemical sensor for detecting Escherichia coli O157:H7 (E. coli O157:H7). The sensor employs a dual-recognition strategy that combines a bacteria-imprinted polymer (BIP) and concanavalin A (ConA). The BIP is formed in situ on the electrode surface as the capture probe, while gold nanoparticles co-functionalized with ConA and the electroactive molecule 6-(ferrocenyl)hexanethiol (Au@Fc-ConA) serve as the signaling probe. When E. coli O157:H7 is present, the bacteria are selectively captured by the BIP. The captured bacteria interact with Au@Fc-ConA through ConA’s sugar-binding properties, triggering Fc oxidation and generating a current proportional to the bacterial concentration. The sensor exhibits a linear detection range of 10¹–10⁵ CFU mL⁻¹ and a low detection limit of 10 CFU mL⁻¹. Additionally, it demonstrates high sensitivity in complex milk samples, detecting E. coli O157:H7 at concentrations as low as 10 CFU mL⁻¹, with recoveries ranging from 94.16% to 110.6%. Even in the presence of a 100-fold higher concentration of E. coli O6, the sensor effectively distinguishes E. coli O157:H7 from it. Moreover, it exhibits high reproducibility with a relative standard deviation of 2%. This study proposes a unique dual recognition strategy that combines simplicity and high performance. This method enables the selective detection of E. coli O157:H7 in real samples, providing a promising tool for food safety monitoring. Full article

Interferon-γ (IFN-γ), a member of the Type II IFN family, is a crucial cytokine in the immune system and serves as an important indicator of immune response. Intracellular cytokine staining (ICS) is a technique used to analyze the production of cytokines within individual cells, and it has a wide range of applications in the fields of immunological monitoring, vaccine trials, and the study of infectious diseases. This study aimed to prepare monoclonal antibodies against duck IFN-γ protein and to establish an ICS protocol for detecting the duck IFN-γ protein. The duIFN-γ-His or duIFN-γ-Fc gene was cloned into the pEE12.4 expression vector and expressed as a recombinant protein of size 20.2 KDa or 54.9 KDa in 293F cells. The purified recombinant proteins were inoculated into BALB/c mice to generate splenic lymphocytes capable of secreting anti-duIFN-γ antibodies, and hybridoma cells were obtained after fusion with SP2/0 cells. A new hybridoma cell line named 24H4, which stably secreted IgG3 κ subtype antibody against duck IFN-γ, was established. This monoclonal antibody (mAb) was identified by Western blot to recognize duck IFN-γ antibodies, and the indirect ELISA results showed that its ability to recognize IFN-γ protein reached 0.001 μg/mL. The established ICS method was used to stain PBMCs after Concanavalin A (ConA) stimulation, and duck IFN-γ protein was successfully detected by flow cytometry, indicating that the ICS method was successful. In this study, we provide a crucial tool for subsequent research on duck cellular immune responses by using the monoclonal antibody 24H4. Full article

Galectin-1 is implicated in several pro-tumourigenic mechanisms and is considered immune-suppressive. The pharmacological inhibition of galectin-1 may be beneficial in cancers in which galectin-1 is overexpressed and driving cancer progression. This study aimed to further characterise the immunosuppressive cytokines influenced by galectin-1 in in vitro immune cell cultures and an in vivo inflammatory model using a recently discovered selective inhibitor of galectin-1, GB1908. To enable a translational approach and link mouse and human pharmacology, anti-CD3/anti-CD28 stimulated T cells cultured from human whole blood and mouse spleens were compared. For in vivo studies of T cell-mediated inflammation, the concanavalin-A (Con-A) mouse model was used to induce a T lymphocyte-driven acute liver injury phenotype. The inhibition of galectin-1 with GB1908 reduced IL-17A, IFNγ and TNFα in a concentration-dependent manner in both mouse and human T cells in vitro. The immunosuppressive cytokines measured in Con-A-treated mice were all upregulated compared to naïve mice. Subsequently, mice treated with GB1908 demonstrated a significant reduction in IL-17A, IFNγ, IL-6 and TNFα compared to vehicle-treated mice. In conclusion, galectin-1 induced the production of several important immune-suppressive cytokines from T cells in vitro and in vivo. This result suggests that, in the context of cancer therapy, a selective galectin-1 could be a viable approach as a monotherapy, or in combination with chemotherapeutic agents and/or checkpoint inhibitors, to enhance the numbers and activity of cytotoxic T cells in the tumour microenvironment of high galectin-1 expressing cancers. Full article

Nasal delivery is a non-invasive strategy for effective drug delivery. Nevertheless, in order to promote drug uptake by the nasal mucosa, it is fundamental to increase its residence time in the administration site. To this aim, nano-sized drug delivery systems are widely exploited. Within this context, the commercially available nanoemulsion for parenteral nutrition is a biocompatible, safe and clinically approved vehicle for drug delivery. Furthermore, the nanodroplet surface can be modified via a well-established protocol to graft Concavalin A, a lectin capable of improving the mucosal adhesion, by binding to the α-mannose and α-glucose residues of the mucosal glycocalyx. The obtained targeted formulation is able to induce haemagglutination, as opposite to non-modified nanoemulsion. Furthermore, the ConA grafting maintains the physicochemical properties of the nanodroplets (size~230 nm, Z < −35 mV) and does not interfere with the loading of the Rose Bengal fluorescent probe. Fluorescently labelled ConA grafted nanodroplets showed enhanced permeation and accumulation in ex vivo bovine nasal mucosa. This study is a proof of concept that Concanavalin A can be used to decorate the surface of nanodroplets, acting as a permeation promoter. Full article

Carbazole is an aromatic heterocyclic organic compound consisting of two fused benzene rings and a pyrrole ring and is a very valuable building structure for the design of many compounds for use in various fields of chemistry and medicine. This study presents three new carbazole-based thiazole derivatives that differ in the presence of a different halogen atom: chlorine, bromine, and fluorine. Experimental studies and quantum-chemical simulations show the effect of changing a halogen atom on the physicochemical, biological, and linear and nonlinear optical properties. We have also found that carbazoles C-Cl, C-Br, and C-F exhibit high tyrosinase inhibitory activity, with IC₅₀ values in the range of 68–105 µM with mixed mechanism of action. Finally, molecular docking to the active site of Concanavalin A (ConA) and bioavailability for all compounds were evaluated. Full article

Bacterial lipopolysaccharides (LPSs) are important indicators of a bacteria presence in any samples. They can therefore be used for the detection of microbiological contamination in food and dairy products. We performed a comparative analysis of different bacterial models by the application of liposomes containing LPS from Salmonella enterica serotype typhimurium on the surface of an 11-mercaptoundecanoic acid (MUA) monolayer chemisorbed on the gold surface of quartz crystal. Using quartz crystal microbalance with dissipation monitoring (QCM-D), we were able to monitor the formation of the lectin, concanavalin A (ConA), layer on the MUA surface. We determined the optimal concentration of the ConA for the layer formation. ConA of 0.3 mg/mL was selected as the most suitable adsorption of liposomes containing LPS. Using the Sauerbrey equation, we calculated that approximately 1.13 × 10¹² ConA molecules per cm² was adsorbed on the MUA surface, which closely corresponds to the 1.19 × 10¹² molecules per cm² by theoretical models. Later, mixed LPS liposomes containing dipalmitoyl phosphatidyl choline (DPPC), dipalmitoyl phosphatidyl ethanolamine (DPPE) and cholesterol successfully interacted with the ConA layer, which resulted in a decrease in the resonant frequency and an increase in dissipation. We compared the adsorption of liposomes with different fractions of LPS and containing LPS from different bacteria. Lack of any LPS in liposomes caused weaker adsorption on the ConA layer. Liposomes containing 50% LPS caused the most prominent adsorption and were suitable for interaction with DNA aptamers specific to certain LPS. The addition of the aptamers to the surface of ConA covered by LPS-containing liposomes resulted in a decrease in resonant frequency and an increase in the dissipation. Using the Kelvin–Voigt viscoelastic model and multiharmonic response of acoustic sensors, we also determined changes in viscoelastic values of the molecular films during interaction with liposomes and the ConA layer. Full article

Recently, the uses of essential oils (EOs) as rumen modifiers, anti-inflammatory agents, and antioxidants were demonstrated in livestock. In the present study, the role of Mentha x piperita L. (MEO), Rosmarinus officinalis L. (REO), and Lavandula angustifolia L. (LEO) EOs in an in vitro sheep model of inflammation was investigated. With this aim, peripheral blood mononuclear cells (PBMCs) were treated with incremental concentrations (3, 5, 7, and 10%) of each EO to test their effects on cell viability and proliferation and on interleukin (IL)-6, IL-10, and IL-8 secretion. The PBMCs were stimulated by Concanavalin A (ConA) alone or in combination with lipopolysaccharide (LPS) mitogen. The positive and negative controls were represented by PBMCs in the presence or absence, respectively, of mitogens only. The cell viability and proliferation were determined by XTT and BrdU assays, while the cytokines were analyzed by ELISA. The EO treatments did not affect the viability; on the contrary, the PBMC proliferation increased in presence of all the EOs tested, according to the different percentages and mitogens used. The IL-10 secretion was higher in both the REO and the LEO tested at 3% than in the positive control; furthermore, the IL-8 level was influenced differently by the various EOs. The present data demonstrate that EOs may modulate the immune response activated by inflammation. Full article

The optimal treatment of diabetes (in particular, type 1 diabetes—T1D) remains a challenge. Closed-loop systems (implants/inserts) provide significant advantages for glucose responsivity and providing real-time sustained release of rapid-acting insulin. Concanavalin A (ConA), a glucose affinity agent, has been used to design closed-loop insulin delivery systems but not without significant risk of leakage of ConA from the matrices and poor mechanical strength of the hydrogels impacting longevity and control of insulin release. Therefore, this work focused on employing a thermoresponsive co-forming matrix between Pluronic F-127 (PL) and structurally robust chitosan (CHT) via EDC/NHS coupling (i.e., covalent linkage of -NH₂ from CHT and ConA to the -COOH of PL). The system was characterized for its chemical structure stability and integrity (FTIR, XRD and TGA), injectability, rheological parameters and hydrogel morphology (Texture Analysis, Elastosens TM Bio2 and SEM). The prepared hydrogels demonstrated shear-thinning for injectability with a maximum force of 4.9 ± 8.3 N in a 26G needle with sol–gel transitioning from 25 to 38 °C. The apparent yield stress value of the hydrogel was determined to be 67.47 Pa. The insulin loading efficiency within the hydrogel matrix was calculated to be 46.8%. Insulin release studies revealed glucose responsiveness in simulated glycemic media (4 and 10 mg/mL) over 7 days (97%) (305 nm via fluorescence spectrophotometry). The MTT studies were performed over 72 h on RIN-5F pancreatic cells with viability results >80%. Results revealed that the thermoresponsive hydrogel is a promising alternative to current closed-loop insulin delivery systems. Full article

Research in the treatment of type 1 diabetes has been addressed into two main areas: the development of “intelligent insulins” capable of auto-regulating their own levels according to glucose concentrations, or the exploitation of artificial intelligence (AI) and its learning capacity, to provide decision support systems to improve automated insulin therapy. This review aims to provide a synthetic overview of the current state of these two research areas, providing an outline of the latest development in the search for “intelligent insulins,” and the results of new and promising advances in the use of artificial intelligence to regulate automated insulin infusion and glucose control. The future of insulin treatment in type 1 diabetes appears promising with AI, with research nearly reaching the possibility of finally having a “closed-loop” artificial pancreas. Full article