Search Results (4,126)

The exogenous application of oxalic acid is a potential approach to amplifying phytoremediation performance on Cd-contaminated soils. However, few studies explore the optimal oxalic acid application regime from a perspective of coupling different concentrations and timings to maximize Cd removal rate. Given this, a pot experiment was conducted using oil sunflower (Helianthus annuus L.) as the test plant. Oxalic acid was added to the pots at concentrations of 1, 2, 3, 4, 5, and 6 mmol/kg at 20, 30, 40, and 50 days after emergence. A control (CK) without exogenous oxalic acid was also included. We examined the discrepancies in various soil Cd forms, sunflower height, plant non-protein thiol (NPT) levels, and soil Cd remediation efficiency under different oxalic acid application regimes. The results showed that applying oxalic acid at a concentration of 4 mmol/kg reduced the proportion of Fe-Mn oxide Cd and organic Cd compared to the control (CK), while increased the proportion of available Cd. The optimal application time is 30 or 40 days after emergence. The addition of exogenous oxalic acid promoted the growth of sunflowers, with the greatest increase in plant height observed when 4 mmol/kg oxalic acid was applied at 30 days after emergence. Exogenous oxalic acid enhanced the absorption of Cd by sunflower roots, with the total Cd accumulation in roots, stems, and leaves being higher than in the control (CK). When 4 mmol/kg oxalic acid was applied at 30 days after emergence, the total Cd accumulation in roots, stems, and leaves was highest. Under different application times and concentration levels of oxalic acid, Cd accumulation was highest in roots, followed by leaves, with stems showing the lowest accumulation. The NPT content in each part is as follows: root > stem > leaf. Applying 5 mmol/kg oxalic acid after 30 days of sunflower emergence resulted in relatively higher total NPT content in roots, stems, and leaves compared to the control (CK). The TOPSIS model was used for comprehensive evaluation, which showed that 4 mmol/kg oxalic acid application at 30 days after emergence could be used as the optimal oxalic acid application regime for phytoremediation. These findings indicate that the addition of oxalic acid effectively promoted the absorption of Cd by sunflower and increased the efficiency of Cd removal from the rhizosphere soil, with the optimal removal of soil Cd achieved by applying oxalic acid at a concentration of 4 mmol/kg 30 days after the emergence of oilseed sunflower seedlings. Full article

This study characterized the binding mechanisms of the lectin cMoL (from Moringa oleifera seeds) to carbohydrates using spectroscopy and molecular dynamics (MD). The interaction with carbohydrates was studied by evaluating lectin fluorescence emission after titration with glucose or galactose (2.0–11 mM). The Stern–Volmer constant (Ksv), binding constant (Ka), Gibbs free energy (∆G), and Hill coefficient were calculated. After the urea-induced denaturation of cMoL, evaluations were performed using fluorescence spectroscopy, circular dichroism (CD), and hemagglutinating activity (HA) evaluations. The MD simulations were performed using the Amber 20 package. The decrease in Ksv revealed that cMoL interacts with carbohydrates via a static mechanism. The cMoL bound carbohydrates spontaneously (ΔG < 0) and presented a Ka on the order of 10², with high selectivity for glucose. Protein–ligand complexes were stabilized by hydrogen bonds and hydrophobic interactions. The Hill parameter (h~2) indicated that the binding occurs through the cMoL dimer. The loss of HA at urea concentrations at which the fluorescence and CD spectra indicated protein monomerization confirmed these results. The MD simulations revealed that glucose bound to the large cavity formed between the monomers. In conclusion, the biotechnological application of cMoL lectin requires specific methods or media to improve its dimeric protein structure. Full article

Electrospun regenerated cellulose (RC) nanofiber membranes were prepared starting from cellulose acetate (CA) with different degrees of substitution. The process was optimized to obtain continuous and uniformly sized CA fibers. After electrospinning, the CA membranes were heat-treated to increase their tensile strength before deacetylation to obtain regenerated cellulose (RC). Affinity membranes were obtained by functionalization, exploiting the hydroxyl groups on the cellulose backbone. 1,4-Butanediol-diglycidyl ether was used to introduce epoxy groups onto the membrane, which was further bioconjugated with the anti-CD63 antibody targeting the tetraspanin CD63 on the extracellular vesicle membrane surface. The highest ligand density was obtained with an anti-CD63 antibody concentration of 6.4 µg/mL when bioconjugation was performed in carbonate buffer. The resulting affinity membrane was tested for the adsorption of extracellular vesicles (EVs) from human platelet lysate, yielding a very promising binding capacity above 10 mg/mL and demonstrating the suitability of this approach. Full article

As a great economic Solanum with ornamental value and good adaptability, Solanum aculeatissimum is considered an excellent candidate for the phytoremediation of Cadmium-contaminated soils. However, there are no studies on the involvement of S. aculeatissimum in the response and tolerance mechanisms of cadmium (Cd) stress. In the present study, S. aculeatissimum was used for the first time for physiological and transcriptomic systematic analysis under different concentrations of Cd stress. The results showed that S. aculeatissimum was indeed well tolerant to Cd and showed Cd enrichment capabilities. Under the Cd stress treatment of 50 mg/kg (Cd6), S. aculeatissimum could still grow normally. At the 90th day of Cd stress, the amount of Cd content in different parts of the plant at the same concentration was in the order of root > stem > leaf. With the extension of the stress time up to 163 d, the trend of Cd content in each part was not consistent, and the results in the root (77.74 mg/kg), stem (30.01 mg/kg), leaf (29.44 mg/kg), immature fruit (18.36 mg/kg), and mature fruit (21.13 mg/kg) of Cd peaked at Cd4, Cd5, Cd1, Cd4, and Cd4, respectively. The enrichment and transport coefficients of all treatments were greater than 1. The treatment groups with the largest and smallest enrichment coefficients were Cd4 and CK, respectively. The treatment groups with the largest and smallest transport coefficients were CK and Cd4, respectively. Malondialdehyde (MDA), peroxidase (POD), and catalase (CAT) in the antioxidant system after Cd stress treatment were significantly increased compared to the untreated group. Under cadmium stress, by using real-time quantitative PCR, four genes (SaHMA20, SaL-AO, SaPrxs4, and SaPCs) were screened for possible correlations to Cd tolerance and absorption enrichment in S. aculeatissimum. The key DEGs are mainly responsible for the metabolic pathways of heavy metal ATPases, plastocyanin protein phytocyanins (PCs), peroxidases (Prxs), and ascorbate oxidase (AAO); these differential genes are believed to play an important role in Cd tolerance and absorption enrichment in S. aculeatissimum. Full article

Background/Objectives: Immune checkpoint inhibitors have an established role in non-small cell lung cancer (NSCLC) therapy. The loss of HLA-class-I expression allows cancer cell evasion from immune surveillance, disease progression, and failure of immunotherapy. The restoration of HLA-class-I expression may prove to be a game-changer in current immunotherapy strategies. Autophagic activity has been recently postulated to repress HLA-class-I expression in cancer cells. Methods: NSCLC cell lines (A549 and H1299) underwent late-stage (chloroquine and bafilomycin) and early-stage autophagy blockage (ULK1 inhibitors and MAP1LC3A silencing). The HLA-class-I expression was assessed with flow cytometry, a Western blot, and RT-PCR. NSCLC tissues were examined for MAP1LC3A and HLA-class-I expression using double immunohistochemistry. CD8+ T-cell cytotoxicity was examined in cancer cells pre-incubated with chloroquine and anti-PD-L1 monoclonal antibodies (Moabs); Results: A striking increase in HLA-class-I expression following incubation with chloroquine, bafilomycin, and IFNγ was noted in A549 and H1299 cancer cells, respectively. This effect was further confirmed in CD133+ cancer stem cells. HLA-class-I, β2-microglobulin, and TAP1 mRNA levels remained stable. Prolonged exposure to chloroquine further enhanced HLA-class-I expression. Similar results were noted following exposure to a ULK1 and a PIKfyve inhibitor. Permanent silencing of the MAP1LC3A gene resulted in enhanced HLA-class-I expression. In immunohistochemistry experiments, double LC3A+/HLA-class-I expression was seldom. Pre-incubation of H1299 cancer cells with chloroquine and anti-PD-L1 MoAbs increased the mean % of apoptotic/necrotic cells from 2.5% to 18.4%; Conclusions: Autophagy blockers acting either at late or early stages of the autophagic process may restore HLA-class-I-mediated antigen presentation, eventually leading to enhanced immunotherapy efficacy. Full article

Seminal extracellular vesicles (SemEVs) are repositories of biomolecules, including metabolites involved in the regulation of sperm function. The correlation between the metabolite profile of SemEVs and semen parameters, along with their role in regulating sperm function, is an unexplored area. This preliminary study evaluated the metabolomic content of SemEVs. Semen samples were obtained from 18 healthy men, and SemEVs were extracted from seminal plasma using the size exclusion chromatography qEV Gen 2–35 nm column coupled with an automatic fraction collector. The physical characterization of SemEVs was carried out with the ZetaView PMX-430-Z QUATT laser system. EV protein markers were detected using Western blot. In addition, these SemEVs were used for metabolomic profiling and functional bioinformatic analysis. The mean concentration of isolated SemEVs was 1.7 ± 1.1 × 10¹¹/mL of seminal plasma, whereas SemEVs size and zeta potential were 129.5 ± 5.5 nm and −40.03 ± 3.99 mV, respectively. Western blot analysis confirmed the presence of EV specific markers such as CD81, ALIX, and TSG101. A total of 107 metabolites were identified using this untargeted metabolomic approach in SemEVs. Bioinformatics analysis further revealed that metabolites associated with tyrosine metabolism were highly enriched in these SemEVs. Ingenuity Pathway Analysis (IPA) also indicated that these metabolites present in SemEVs were involved in the regulation of the free radical scavenging pathway. Furthermore, our metabolomic results suggest that these SemEV-associated metabolites may play a pivotal role in the maintenance of seminal plasma redox homeostasis. Full article

Background: The Peroxidase (PRX) gene family is essential for plant growth and significantly contributes to defense against stresses. However, information about PRX genes in cucumber (Cucumis sativus L.) remains limited. Methods: In this present study, CsPRX genes were identified and characterized using bioinformatics analysis. The expression pattern analysis of CsPRX genes were examined utilizing the RNA-seq data of cucumber from public databases and real-time quantitative PCR (qRT-PCR) analysis. Results: Here, we identified 60 CsPRX genes and mapped them onto seven chromosomes of cucumber. The CsPRX proteins exhibited the presence of 10 conserved motifs, with motif 8, motif 2, motif 5, and motif 3 consistently appearing across all 60 CsPRX protein sequences, indicating the conservation of CsPRX proteins. Furthermore, RNA-seq analysis revealed that differential expression of CsPRX genes in various tissues. Notably, a majority of the CsPRX genes exhibited significantly higher expression levels in the root compared to the other plant tissues, suggesting a potential specialization of these genes in root function. In addition, qRT-PCR analysis for four selected CsPRX genes under different stress conditions indicated that these selected CsPRX genes demonstrated diverse expression levels when subjected to NaCl, CdCl₂, and PEG treatments, and the CsPRX17 gene was significantly induced by NaCl, CdCl₂, and PEG stresses, suggesting a vital role of the CsPRX17 gene in response to environmental stresses. Conclusions: These findings will contribute valuable insights for future research into the functions and regulatory mechanisms associated with CsPRX genes in cucumber. Full article

One of the main objectives of developing new anti-cancer vaccine strategies is to effectively induce CD8+ T cell-mediated anti-tumor immunity. Live recombinant vectors, notably Listeria monocytogenes, have been shown to elicit a robust in vivo CD8+ T-cell response in preclinical settings. Significantly, it has been demonstrated that Listeria induces inflammatory/immunogenic cell death mechanisms such as pyroptosis and necroptosis in immune cells that favorably control immunological responses. Therefore, we postulated that the host’s response to Listeria-based vectors and the subsequent induction of CD8+ T cell-mediated immunity would be compromised by the lack of regulatory or effector molecules involved in pyroptosis or necroptosis. To test our hypothesis, we used recombinant L. monocytogenes carrying the ovalbumin gene (LM.OVA) to vaccinate wild-type (WT), caspase-1/11^−/−, gsdmd^−/−, ripk3^−/−, and mlkl^−/− C57Bl/6 mice. We performed an in vivo cytotoxicity assay to assess the efficacy of OVA-specific CD8+ T lymphocytes in eliminating target cells in wild-type and genetically deficient backgrounds. Furthermore, we evaluated the specific anti-tumor immune response in mice inoculated with the B16F0 and B16F0.OVA melanoma cell lines. Our findings demonstrated that while caspase-1/11 and GSDMD deficiencies interfere with the rapid control of LM.OVA infection, neither of the KOs seems to contribute to the early activation of OVA-specific CTL responses. In contrast, the individual deficiency of each one of these proteins positively impacts the generation of long-lasting effector CD8+ T cells. Full article

Background/Objectives: In this study, serum selenium levels in patients with Crohn’s disease (CD) and ulcerative colitis (UC) were evaluated to identify potential predictive markers of disease activity. Conducted in 100 inflammatory bowel disease (IBD) patients (54 CD, 46 UC) and 100 healthy controls, this research provides novel insights through focusing on the regional selenium status of people with IBD in the Polish population, a demographic with limited existing data. Methods: Selenium concentrations were measured using inductively coupled plasma mass spectrometry (ICP-MS). Results: Significantly lower levels of selenium were observed in CD (64.79 µg/L ± 12.15 µg/L) and UC (68.61 µg/L ± 11.43 µg/L) patients when compared with the controls (90.52 ± 12.00 µg/L, p < 0.0001). Regression analysis identified leukocyte and erythrocyte counts and bilirubin as significant predictors of selenium levels in UC patients, while no significant predictors were found for CD. Conclusions: The findings suggest that selenium deficiency is linked to IBD and may serve as a non-invasive biomarker for disease severity, particularly in UC. This practical approach offers a potential alternative to invasive procedures such as endoscopy for monitoring disease progression. However, further research is needed to confirm these findings in larger populations and explore the therapeutic role of selenium supplementation in IBD management. Full article

It has been shown that the response rate of TNBC is dependent on the level of PD-L1 and the tumor microenvironment (TME). Approaches that alter the TME can improve the efficacy of ICIs. Background: We have engineered a Smac-armed oncolytic virus by inserting a Smac transgene into the genome of a vesicular stomatitis virus to generate VSV-S. Our previous study shows that the anticancer efficacy of VSV-S is more potent than that of wild-typed VSV in a subcutaneous TNBC mouse model. VSV-S treatment reverts the immunosuppressive TME by reducing MDSCs and TAMs, while increasing infiltration of neutrophils and CD8+ T cells. Methods: VSV-S was used to treat TNBC in an orthotopic mouse model, and in a combination therapy with an anti-PD-1 antibody to treat metastatic TNBC in a mouse model. Changes in the TME were evaluated. Results: In this current study, we show that neoadjuvant VSV-S treatment of primary orthotopic TNBC tumors in mice drastically lowered lung metastasis after surgical removal of the primary tumor, and significantly increased the survival rate. The mechanism of action and changes to the TME were delineated, among which one significant marker is the elevation of PD-L1 expression in tumors. In the TNBC lung metastasis mouse model, pulmonary treatment with VSV-S greatly enhanced the efficacy of ICI treatment. Conclusions: Our results suggest that the combination of oncolytic virus and ICI therapies has the potential to substantially improve the outcome of TNBC treatment. Full article

Neuropathic pain (NPP) is a devastating and unbearable painful condition. As prevailing treatment strategies have failed to mitigate its complications, there remains a demand for effective therapies. Electroacupuncture (EA) has proved a potent remedial strategy in NPP management in humans and mammals. However, past studies have investigated the underlying mechanism of the analgesic effects of EA on NPP, focusing primarily on adenosine receptors in peripheral tissues. Herein, we elucidate the role of the adenosine (Adora-3) signaling pathway in mediating pain relief through EA in the central nervous system, which is obscure in the literature and needs exploration. Specific pathogen-free (SPF) male adult mice (C57BL/6 J) were utilized to investigate the effect of EA on adenosine metabolism (CD73, ADA) and its receptor activation (Adora-3), as potential mechanisms to mitigate NPP in the central nervous system. NPP was induced via spared nerve injury (SNI). EA treatment was administered seven times post-SNI surgery, and lumber (L4–L6) spinal cord was collected to determine the molecular expression of mRNA and protein levels. In the spinal cord of mice, following EA application, the expression results revealed that EA upregulated (p < 0.05) Adora-3 and CD73 by inhibiting ADA expression. In addition, EA triggered the release of adenosine (ADO), which modulated the nociceptive responses and enhanced neuronal activation. Meanwhile, the interplay between ADO levels and EA-induced antinociception, using an Adora-3 agonist and antagonist, showed that the Adora-3 agonist IB-MECA significantly increased (p < 0.05) nociceptive thresholds and expression levels. In contrast, the antagonist MRS1523 exacerbated neuropathic pain. Furthermore, an upregulated effect of EA on Adora-3 expression was inferred when the Adora-3 antagonist was administered, and the EA treatment increased the fluorescent intensity of Adora-3 in the spinal cord. Taken together, EA effectively modulates NPP by regulating the Adora-3 signaling pathway under induced pain conditions. These findings enhance our understanding of NPP management and offer potential avenues for innovative therapeutic interventions. Full article

The dwindling availability of agricultural land, caused by factors such as rapid population growth, urban expansion, and soil contamination, has significantly increased the pressure on food production. To address this challenge, cultivating non-food crops on contaminated land has emerged as a promising solution. This approach not only frees up fertile soil for food production but also mitigates human exposure to contaminants. This work aimed to examine the impact of soil contamination with Cd, Pb, Ni, and Zn on the growth, productivity, metal accumulation, and the tolerance of five lignocellulosic non-food crops: switchgrass (Panicum virgatum L.), biomass sorghum (Sorghum bicolor L. Moench), giant reed (Arundo donax L.), African fodder cane (Saccharum spontaneum L. spp. aegyptiacum Willd. Hackel), and miscanthus (Miscanthus × giganteus Greef et Deu.). A two-year pot experiment was conducted in Greece, Italy, and Portugal, following the same protocols and applying various levels of metals: Cd (0, 4, 8 mg kg⁻¹), Pb and Zn (0, 450, 900 mg kg⁻¹), and Ni (0, 110, 220 mg kg⁻¹). The experimental design was completely randomized, with three replicates for each treatment. The results showed that switchgrass and sorghum generally maintained their height and productivity under Cd and Pb stress but were adversely affected by high Zn and Ni concentrations. Giant reed and African fodder cane showed reduced height and productivity at higher Ni and Zn levels. Miscanthus exhibited resilience in height but experienced productivity reductions only at the highest Zn concentration. Heavy metal uptake varied among crops, with switchgrass and sorghum showing high Cd and Pb uptake, while giant reed accumulated the most Cd and Zn. Miscanthus had the highest Ni accumulation. The tolerance indices indicated that switchgrass and sorghum were more tolerant to Cd and Zn at lower concentrations, whereas miscanthus had lower tolerance to Cd but a higher tolerance to Zn at higher concentrations. Giant reed and African fodder cane demonstrated stable tolerance across most heavy metals. Accumulation indices highlighted the effectiveness of switchgrass and sorghum in Cd and Pb uptake, while miscanthus excelled in Ni and Zn accumulation. The cluster analysis revealed similar responses to heavy metal stress between African fodder cane and giant reed, as well as between sorghum and miscanthus, with switchgrass displaying distinct behavior. Overall, the study highlights the differential tolerance and accumulation capacities of these crops, indicating the potential for phytoremediation applications and biomass production in heavy metal-contaminated soils. Full article

Greek giant beans, also known as “Gigantes Elefantes” (elephant beans, Phaseolus vulgaris L.,) are a traditional and highly cherished culinary delight in Greek cuisine, contributing significantly to the economic prosperity of local producers. However, the issue of food fraud associated with these products poses substantial risks to both consumer safety and economic stability. In the present study, multi-elemental analysis combined with decision tree learning algorithms were investigated for their potential to determine the multi-elemental profile and discriminate the origin of beans collected from the two geographical areas. Ensuring the authenticity of agricultural products is increasingly crucial in the global food industry, particularly in the fight against food fraud, which poses significant risks to consumer safety and economic stability. To ascertain this, an extensive multi-elemental analysis (Ag, Al, As, B, Ba, Be, Ca, Cd, Co, Cr, Cs, Cu, Fe, Ga, Ge, K, Li, Mg, Mn, Mo, Na, Nb, Ni, P, Pb, Rb, Re, Se, Sr, Ta, Ti, Tl, U, V, W, Zn, and Zr) was performed using Inductively Coupled Plasma Mass Spectrometry (ICP-MS). Bean samples originating from Kastoria and Prespes (products with Protected Geographical Indication (PGI) status) were studied, focusing on the determination of elemental profiles or fingerprints, which are directly related to the geographical origin of the growing area. In this study, we employed a decision tree algorithm to classify Greek “Gigantes Elefantes” beans based on their multi-elemental composition, achieving high performance metrics, including an accuracy of 92.86%, sensitivity of 87.50%, and specificity of 96.88%. These results demonstrate the model’s effectiveness in accurately distinguishing beans from different geographical regions based on their elemental profiles. The trained model accomplished the discrimination of Greek “Gigantes Elefantes” beans from Kastoria and Prespes, with remarkable accuracy, based on their multi-elemental composition. Full article

This study investigated the effects of pure and methyl-β-cyclodextrin loaded forms of resveratrol (10 µg/mL, 20 µg/mL, and 40 µg/mL) on ram sperm functions post-thawing. Semen samples were pooled and divided into ten groups: Control, RES10, RES20, RES40, CD10, CD20, CD40, RLC10, RLC20, and RLC40. The groups were pre-diluted with media containing the group-specific chemicals, followed by 15 min of incubation, dilution, and freezing. To assess the effects of the chemicals, a post-thaw sperm quality assessment was conducted. Motility and other velocity parameters were evaluated using computer-assisted semen analysis. The functional integrity of spermatozoa membranes was assessed with the hypo-osmotic swelling test, and the capacitation status of spermatozoa was determined through fluorescent microscopic evaluation. Additionally, flow cytometry was used to evaluate mitochondrial activity, oxidative stress, and the integrity of the sperm membrane and acrosome. The results indicated that cyclodextrin adversely affected sperm functions following freezing–thawing, notably increasing the rate of spermatozoa exhibiting pre-capacitation and mitochondrial activity by approximately 34% and 16%, respectively (p < 0.05). It was found that 20 µg/mL resveratrol prevented pre-capacitation (p < 0.05). Both resveratrol and resveratrol-loaded cyclodextrin groups improved post-thaw sperm qualities overall, demonstrating their utility for freezing ram semen. However, higher concentrations of resveratrol were found to negatively impact sperm functions. Full article

Heavy metals are one of the main threats to marine life and ecosystems and any remedial action in that regard is urgently required. The aim of this work is to study the bioremoval of cadmium, chromium and lead in a microalgae strain Chlorococcum dorsiventrale isolated from Tunisian coastal waters along with assessing its enzymatic and molecular responses. The microalgae were tested in artificial seawater to evaluate their capacity for phycoremediation in an aquatic environment. This strain tolerated exposure to Cd (II), Cr (VI), and Pb (II) and was able to grow for 14 days. Cd and Cr exposures elicited a decrease in chlorophyll, lipid and polysaccharide contents, whereas no damages were detected following Pb treatment. For protein content, no significant changes were seen except after Pb exposure which induced a slight increase after treatment with 5 mg/L. The assessment of stress defense-related gene expression using qRT-PCR revealed that exposure to Pb and Cr induced an up-regulation of catalase, superoxide dismutase and photosystem II protein D1 encoding genes. Moreover, heat shock protein 70 was slightly overexpressed. Removal efficiencies for Cr and Pb attained 89% and 95%, respectively. The mechanisms by which C. dorsiventrale removed Cr involved both intracellular and extracellular biosorption, while Pb was predominantly removed through membrane adsorption. This study highlights the potential of C. dorsiventrale as an efficient agent for the bioremediation of heavy metal-contaminated water, including industrial wastewater, thus paving the way for practical and environmental applications in pollution control. Full article