Search Results (32,465)

This research employed “Response Surface Methodology (RSM)” to assess the effectiveness of electrocoagulation (EC) in treating olive mill wastewater (OMW) before applying adsorption with olive stone biochar (OS) as a sustainable adsorbent. Several parameters, including reaction time, current density (CD), inter-electrode distance, and the number of electrodes, were optimized. Analysis using Minitab 22.2 resulted in robust regression models with high coefficients of determination (R²). The optimal parameters were CD of 12.41 mA/cm², a time of 45.61 min, an inter-electrode spacing of 1 cm, and a maximum of 6 electrodes, resulting in an energy consumption (ENC) of 9.85 kWh/m³. Significant pollutant percentage removals were achieved: 72.32% for total Kjeldahl nitrogen (TKN), 80.74% for turbidity, 57.44% for total phenol (TPh), 56.9% for soluble chemical oxygen demand (COD_soluble), and 56.6% for total chemical oxygen demand (COD_total). After the EC, the adsorption of pollutants was conducted using OS biochar that was generated through the pyrolysis of OS at a temperature of 500 °C. FTIR analysis of the biochar revealed key absorption bands that indicated the presence of inorganic compounds, aromatic C=C, and phenolic groups O-H. The integrated EC and adsorption (ECA) process demonstrated markedly higher efficiencies, with TPh removal reaching 61.41%, turbidity reduction at 81.92%, TKN reduction at 77.78%, COD_soluble reduction at 70.31%, COD_total reduction at 65.1%, and project cost of $2.88/m³. The ECA process presents a promising treatment approach for OMW. Full article

In mammals, CD22 is a member of the Siglec family and plays essential roles in B-cell activation, signal transduction, and immune regulation. However, its functions in teleost fish remain largely unclear. In this study, a CD22 homolog designated On-CD22 was identified and cloned from Nile tilapia (Oreochromis niloticus). On-CD22 transcripts were highly expressed in the head kidney and peripheral blood of healthy fish and showed significant expression changes following infection with Streptococcus agalactiae, Aeromonas hydrophila, or stimulation with poly(I:C). Subcellular localization analysis indicated that On-CD22 is predominantly localized to the plasma membrane. Luciferase reporter assays performed in heterologous cell systems showed that overexpression of On-CD22 was associated with changes in the basal transcriptional activities of NF-κB, IFN1, IFN3, and STAT1 responsive promoters under unstimulated conditions. Furthermore, single-cell transcriptomic analysis revealed that On-CD22 expression was mainly confined to the B-cell population within head-kidney leukocytes. Collectively, these findings suggest that On-CD22 may be involved in immune regulatory processes in Nile tilapia. Full article

Background: Depression and Metabolic Dysfunction-Associated Steatotic Fatty Liver Disease (MASLD) are common chronic diseases, respectively. However, the causal and molecular links between them remain unclear. In order to explore whether depression contributes to an increased risk of MASLD and whether inflammation mediates this effect, we integrated multi-level evidence from the epidemiology of the National Health and Nutrition Examination Survey (NHANES), the genetics of GWAS, the transcriptomes of GEO, and single-cell RNA sequencing datasets. Methods: A multi-level integrative analysis strategy was used to validate this pathway. First, a cross-sectional epidemiological analysis based on NHANES data was used to reveal the association between depression and MASLD, and to explore the mediating role of inflammation and liver injury markers. Secondly, a two-sample Mendelian randomization analysis was used to infer the causal direction of depression and MASLD, and to verify the mediating effect of systemic inflammation and liver injury indicators at the genetic level. Then, the transcriptome co-expression network analysis and machine learning were used to screen the common hub genes connecting the two diseases. Finally, single-cell transcriptome data were used to characterize the dynamic expression of potential key genes during disease progression at cellular resolution. Results: Depression significantly increased the risk of MASLD, especially in women (OR = 1.39, 95%CI [1.17–1.65]). Parallel mediation analysis showed that high-sensitivity C-reactive protein (hs-CRP) (p < 0.001), γ-glutamyltransferase (GGT) (p < 0.001), and alkaline phosphatase (ALP) (p < 0.001) mediated this relationship. Mendelian randomization analysis confirmed the unidirectional causal effect of depression on MASLD, and there was no reverse association (β = 0.483, SE = 0.146, p = 0.001). Weighted gene co-expression network analysis and machine learning identified CD40LG as a potential molecular bridge between depression-associated immune modules and MASLD. In addition, single-cell data analysis revealed a stage-specific trend of CD40LG expression in CD4⁺ T cells during MASLD progression, while its receptor CD40 was also activated in B cells. In the female sample, CD40LG maintained an upward trend. However, the stability of this result is limited by the limited sample size. Conclusions: This study provides converging multi-omics evidence that depression plays a causal role in MASLD through inflammation-mediated immune signaling. The CD40LG-CD40 axis has emerged as an immune mechanism that transposes depression into the pathogenesis of MASLD, providing a potential target for the intervention of gender-specific metabolic liver disease. Full article

Chimeric antigen receptor (CAR)-T-cell therapy is a revolutionary approach in immunotherapy that has shown remarkable success in the treatment of blood cancers. Many preclinical studies are currently underway worldwide to extend the CAR-T-cell therapy benefits to a broad spectrum of cancers, using rodent models. Alternative in vivo platforms are essential for overcoming the drawbacks associated with rodent models, including immunodeficiency in humanized models, ethical concerns, extended time requirements, and cost. In this work, we used the chicken chorioallantoic membrane (CAM) assay to evaluate the in vivo efficacy of cluster-of-differentiation 19 (CD19)-targeting CAR-T cells expressing a second-generation CAR construct against human lymphoma derived from the Raji cell line. Our results confirm the efficacy of selected CAR-T cells on tumor growth, metastasis, and angiogenesis. Further, the chicken embryo has an intrinsic active immune system. Therefore, the dialog between CAR-T cells and endogenous immune cells, as well as their participation in the tumor challenge, has also been studied. In conclusion, our study demonstrates that the chicken CAM assay provides a relevant in vivo, 3Rs (Replacement, Reduction and Refinement)-compliant new approach methodology (NAM), which is well-suited for the current needs of preclinical research on CAR-T-cell therapy. Full article

Lentiviral transduction remains the gold standard in adoptive modified cellular therapy, such as CAR-T; however, genome integration is not always desirable, such as when treating non-fatal autoimmune disease or for additional editing steps using CRISPR to produce allogeneic CAR-modified cells. Delivering in vitro-transcribed (IVT) mRNA represents an alternative solution but the labile nature of mRNA has led to efforts to improve half-life and translation efficiencies using a range of approaches including chemical and structural modifications. In this study, we explore the role of N⁶–methyladenosine (m6A) in a CD19-CAR sequence when delivered to T cells as an IVT mRNA. In silico analysis predicted the presence of four m6A consensus (DRACH) motifs in the CAR coding sequence and treating T cells with an inhibitor of the m6A methyltransferase (METTL3) resulted in a significant reduction in CAR protein expression. RNA analysis confirmed m6A bases at three of the predicted sites, indicating that the modification occurs independently of nuclear transcription. Synonymous mutation of the DRACH sites reduced the levels of CAR protein from 15 to >50% depending on the T cell donor. We also tested a panel of CAR transcripts with different UTRs, some containing m6A consensus motifs, and identified those which further improved protein expression. Furthermore, we found that the methylation of consensus m6A sites seems to be somewhat sequence-context-dependent. These findings demonstrate the importance of the m6A modification in stabilising and enhancing expression from IVT-derived mRNA and that this occurs within the cell, meaning targeted in vitro chemical modification during mRNA manufacturing may not be necessary. Full article

In type 1 diabetes (T1D) in non-obese diabetic (NOD) mice, dendritic cells (DCs) exhibit a Stat5b mutation that impairs regulatory T cell (Tregs) numbers and suppressive function. To correct this defect, we generated transgenic NOD mice expressing constitutively active Stat5b (NOD.Stat5b-CA) in DCs, which conferred protection from diabetes that was associated with an expanded Treg population and a marked reduction in CD8⁺ T cell frequencies in secondary lymphoid organs. However, the phenotypic characteristics and underlying mechanisms to eliminate CD8⁺ T cells in NOD.Stat5b-CA mice are unknown. In this study, we found that the frequency of Tregs was significantly higher in the thymus and peripheral lymphoid organs of NOD.Stat5b-CA mice compared with NOD mice. Tregs in the peripheral lymphoid organs exhibited increased expression of activation markers CD69 and OX40, alongside reduced CD62L. We also found that CD8⁺ T cell frequencies were reduced in the peripheral organs but not in the thymus of NOD.Stat5b-CA mice, while CD4⁺ T cell frequencies remained unchanged across all organs. Furthermore, NOD.Stat5b-CA mice exhibited a reduced frequency of central Tregs (CD62L^high CD44^low) and increased frequency of effector Tregs (CD62L^low CD44^high) under steady-state conditions compared to NOD mice. Notably, Tregs from NOD.Stat5b-CA mice displayed enhanced cytotoxic activity, evidenced by increased expression of perforin, granzyme B, and Fas ligand, potentially mediating CD8⁺ T cell frequency reduction. Collectively, these findings highlight a novel role for Stat5b-CA.DC-educated Tregs in modulating immune responses by eliminating peripheral pathogenic CD8⁺ T cells via cytotoxic pathways, thereby contributing to immune regulation in NOD.Stat5b-CA mice. Full article

Multiple Myeloma (MM) is an incurable malignancy that progresses from asymptomatic precursor stages—Monoclonal Gammopathy of Undetermined Significance (MGUS) and Smouldering Multiple Myeloma (SMM)—to active disease. Despite ongoing research, the molecular mechanisms driving this progression remain poorly understood. In this study, we aimed to uncover key regulatory factors involved in MM progression by integrating single-cell RNA sequencing (scRNA-seq) data with curated a priori biological knowledge of MM. To this end, we first integrated a priori knowledge from databases in a synthetic gene network map to play the role of an MM-related backbone to project findings from scRNA analysis on CD138⁺ Plasma Cells. This was followed by stage-specific regulatory network construction and analysis using Integrated Value of Influence (IVI) metrics to identify the most influential genes across disease stages. Our findings revealed GSK3B, RELA, CDKN1A, and PCK2 as central regulators shared across multiple stages of the disease. Notably, several of these genes had not previously been included in established MM gene sets, highlighting them as prime candidates for biomarkers and drug targets. Full article

Objectives: Clinical decision-support systems are computer-based tools to improve healthcare decision-making. However, their effectiveness depends on being positively perceived and well understood by healthcare professionals. Qualitative research is particularly valuable for exploring related behaviors and attitudes. This study aims to explore experiences of family physicians and nurses concerning the visualization, utility and understanding of the non-valvular atrial fibrillation clinical decision-support system (CDS-NVAF) tool in primary care in Catalonia, Spain. Methods: We performed a qualitative study, taking a pragmatic utilitarian approach, comprising focus groups with healthcare professionals from primary care centers in the intervention arm of the CDS-NVAF tool randomized clinical trial. A thematic content analysis was performed. Results: Thirty-three healthcare professionals participated in three focus groups. We identified three key themes: (1) barriers to tool adherence, encompassing problems related to understanding the CDS-NVAF tool, alert fatigue, and workload; (2) using the CDS-NVAF tool: differences in interpretations of Time in Therapeutic Range (TTR) assessments, and the value of TTR for assessing patient risk; (3) participants’ suggestions: improvements in workflow, technical aspects, and training in non-valvular atrial fibrillation management. Conclusions: Healthcare professionals endorsed a clinical decision-support system for managing oral anticoagulation in non-valvular atrial fibrillation patients in primary care. However, they emphasized the view that the CDS-NVAF requires technical changes related to its visualization and better integration in their workflow, as well as continuing training to reinforce their theoretical and practical knowledge for better TTR interpretation. Full article

Electron transfer (ET) is a foundational biogeochemical process in paddy soils, distinctively molded by alternating anaerobic-aerobic conditions from flooding-drainage cycles. Despite extensive research on heavy metal(loid) (denoted as “HM”, e.g., As, Cd, Cr, Hg) dynamics in paddies, ET has not been systematically synthesized as a unifying regulatory mechanism, and the trade-offs of ET-based mitigation strategies remain unclear. These critical gaps have drastically controlled HMs’ mobility, which further modulates bioavailability and subsequent accumulation in rice (Oryza sativa L., a staple sustaining half the global population), posing substantial food safety risks. Alongside progress in electroactive microorganism (EAM) research, extracellular electron transfer (EET) mechanism delineation, and soil electrochemical monitoring, ET’s role in orchestrating paddy soil HM dynamics has garnered unparalleled attention. This review explicitly focuses on the linkage between ET processes and HM biogeochemistry in paddy ecosystems: (1) elucidates core ET mechanisms in paddy soils (microbial EET, Fe/Mn/S redox cycling, organic matter-mediated electron shuttling, rice root-associated electron exchange) and their acclimation to flooded conditions; (2) systematically unravels how ET drives HM valence transformation (e.g., As(V) to As(III), Cr(VI) to Cr(III)), speciation shifts (e.g., exchangeable Cd to oxide-bound Cd), and mobility changes; (3) expounds on ET-regulated HM bioavailability by modulating soil retention capacity and iron plaque formation; (4) synopsizes ET-modulated HM accumulation pathways in rice (root uptake, xylem/phloem translocation, grain sequestration); (5) evaluates key factors (water management, fertilization, straw return) impacting ET efficiency and associated HM risks. Ultimately, we put forward future avenues for ET-based mitigation strategies to uphold rice safety and paddy soil sustainability. Full article

This study successfully developed an oral vaccine for Type 1 Diabetes utilizing recombinant Lactococcus lactis expressing the GAD65 autoantigen. We conducted an in-depth investigation into its protective mechanisms in NOD mice, with a particular focus on its effects on the gut microbiota and metabolome. The administration of the GAD65-L. lactis vaccine resulted in a significant delay in diabetes onset and the preservation of pancreatic function. Our analyses revealed notable alterations in the gut microbial ecosystem, enhancing its diversity and the abundance of beneficial bacteria. Metabolomic profiling indicated time-dependent changes in metabolic pathways, with a marked enrichment of pyrimidine metabolism at 16 weeks and arachidonic acid metabolism at 24 weeks after vaccination by both GAD65-L. lactis and NZ9000-L. lactis. Integrated correlation analysis identified specific microbiota–metabolite interactions, including associations between Ruminiclostridium and lipid species in the GAD65-L. lactis group. These modifications in the microbial community and metabolic landscape were accompanied by enhanced immunoregulatory responses in intestinal LPLs, including expanded Treg populations and suppressed CD8⁺ T cells, a rising trend in IL-10-producing naive dendritic cells, and increased concentrations of TGF-β. Full article

Background and Objectives: Basal cell carcinoma (BCC) is the most common skin carcinoma, mainly occurring in older individuals. The aim of this study was to document the immunohistochemical distribution of CD34 in different histopathological types of BCC, as well as in the peritumoral and uninvolved skin of biopsy samples. Materials and Methods: Excisional biopsies of skin BCCs were routinely processed into paraffin blocks, and microtome sections were stained immunohistochemically for CD34. Results: A consistent finding in skin samples containing BCC was the absence of CD34 in the following extravascular structures: neoplastic cells, epidermis and its derivatives (except for the cells of the isthmic part of the outer hair follicle sheath), fibroblast-like cells of BCC tumor stroma, as well as in the papillary dermis in the tumor region. Fibroblast-like cells of the tumor stroma were variably CD34 immunopositive only in the nodular type of BCC. In all examined biopsies, part of the dermis adjacent to the BCC tumor mass (juxtatumoral zone) was characterized by pronounced CD34 immunopositivity. In the transitional zone of peritumoral skin and in marginal skin, CD34-positive connective tissue cells were observed in the periadnexal dermis around: sebaceous gland lobules, the secretory coils of eccrine sweat glands, the pilosebaceous canal, as well as in the perimysium of the arrector pili muscle. Fibrocytes of fibrous sheaths encasing the isthmic part of hair follicles were CD34 negative, interposed between highly positive epithelial cells of the outer hair follicle sheath and the fibroblasts of the local reticular dermis. The transitional zone and uninvolved skin contained CD34-positive fibroblast-like cells situated between secondary bundles of reticular dermis, as well as CD34-positive cell processes within these bundles. Conclusions: The observed pattern of CD34 positivity within the examined regions shows a specific distribution, providing insight into the adaptive responses of the skin to the tumoral process. Full article

Background: Ovarian cancer remains the deadliest gynecologic malignancy, with its progression closely tied to age-associated remodeling of the tumor immune microenvironment. The laying hen serves as a valuable spontaneous model for human ovarian cancer. Its single-cell analyses may provide valuable insights into the immune-related axis linking ovarian aging to carcinogenesis. Methods: This study applied single-cell RNA sequencing to profile ovaries from three laying hen groups, including 35-week-old normal ovaries (A35w), 110-week-old normal ovaries (B110w), and 110-week-old ovarian cancer tissues (C110w). Key analyses had UCell-based scoring of senescence-related pathways and cancer hallmarks, differential expression analysis for overlapping dysregulated genes, LASSO regression-based prognostic model construction, and assessment of chemotherapy sensitivity and immune infiltration. Results: A comprehensive cellular landscape of chicken ovaries was established, identifying major immune populations including B cells, CD4+ T cells, CD8+ T cells, macrophages, and plasma cells. Senescence-related pathways and cancer hallmarks showed progressive activation in immune cells from A35w to B110w to C110w. A total of 216 genes commonly dysregulated in aging and carcinogenesis, reveal core links between immune dysfunction and malignant transformation. The 20-gene prognostic model derived from these genes stratified human ovarian cancer patients into high-risk and low-risk groups with significant overall survival differences, exhibited robust predictive performance across TCGA, GSE32063, and GSE140082. The model also predicted the differential chemotherapy sensitivity in high-risk and low-risk patients and correlated with specific immune infiltration patterns in the tumor microenvironment. Conclusions: Notably, this is the first single-cell RNA sequencing study of chicken ovarian cancer, and we constructed the 20-gene prognostic model for human ovarian cancer using 216 genes that change significantly in immune cells during both ovarian aging and carcinogenesis. This work provides support to establish the hen as a potential preclinical animal model and a translational tool to guide personalized therapy. Full article

This study aims to characterise the B cell compartment in a cohort of Sicilian centenarians by analysing absolute CD3⁻CD19⁺ lymphocyte counts, in association with age, sex, cytomegalovirus (CMV) serostatus, related to immune ageing, and interleukin (IL)-6 levels, representative of inflamm-ageing. It also investigates age-related changes in the CD4⁺/CD19⁺ ratio as a marker of immune ageing, reflecting shifts in immune homeostasis. B cell counts were assessed by flow cytometry on 53 Sicilians aged 19–110 years: 20 Adults, 15 Older adults, 11 long-living individuals, and 7 oldest centenarians. A multiple negative binomial regression was applied to evaluate the effects of age, sex, CMV serostatus, and Il-6 levels on values of B cells. The results showed a non-significant trend toward age-related decline without sex-based differences. A significant reduction in B cell count was observed in individuals with high anti_CMV titres, while IL-6 levels showed a borderline inverse correlation. CD4⁺/CD19⁺ ratio values showed an age-related increase. Our findings suggest that the age-related decline in B cell numbers may be mostly related to CMV infection and IL-6 values, without sex contribution. The age-related increase in the CD4⁺/CD19⁺ ratio, most pronounced in oldest centenarians, may represent a compensatory adaptation promoting immune regulation and chronic inflammation control. Full article

Cutaneous squamous cell carcinoma (cSCC) is an immunogenic malignancy with variable immune infiltration and inconsistent responses to checkpoint blockade. Tumor-infiltrating lymphocytes (TILs) influence tumor progression and therapeutic outcome, yet their phenotypic and functional diversity across disease contexts remains incompletely understood. This review systematically characterizes the TIL landscape in human cSCC. Following PRISMA 2020 guidelines, PubMed and Embase were searched up to May 2025 and restricted to studies evaluating tumor-infiltrating lymphocytes in human cSCC, using the modified Newcatle–Ottawa score to assess risk of bias. Data were synthesized qualitatively given methodological heterogeneity. 48 studies met inclusion criteria. cSCCs exhibited dense CD3⁺ infiltrates composed of cytotoxic (CD8⁺GzmB⁺, Ki-67⁺, CD69⁺) and regulatory (FOXP3⁺, CCR4⁺) subsets. Higher CD8⁺ activity correlated with smaller tumors and longer disease-free survival, whereas FOXP3⁺ enrichment and TGF-β2 signaling promoted immune evasion. Immunosuppressed patients demonstrated diminished CD8⁺ density and clonality. Immune modulation with PD-1/PD-L1 blockade, imiquimod, HPV vaccination, or OX40 stimulation enhanced effector function. The cSCC immune microenvironment reflects a balance between cytotoxic and suppressive factors. Harmonizing multimodal immune profiling and integrating spatial context with systemic immune status may advance both prognostic stratification and therapeutic design. Full article

Urease-producing bacteria (UPB) have great potential for the bioremediation of heavy-metal pollution through biomineralization and adsorption. In this study, a strain of UPB, C7-12, was isolated from heavy-metal-contaminated soil in a lead–zinc mining area and identified as Serratia marcescens. The heavy-metal removal ability, influencing factors, and precipitation mode of this UPB strain in solution were investigated. The cadmium (Cd) removal rate in a Cd (1 mg/L) solution from C7-12 reached 85%, and pH was the main influencing factor. With urea mediation, S. marcescens C7-12 biomineralizes the Cd²⁺ in solution to form CdCO₃ and removes it through extracellular precipitation and surface adsorption. Furthermore, the removal rates of Cd²⁺, Pb²⁺, Zn²⁺ and Cu²⁺ in solution by S. marcescens C7-12 were 33–65%, 28–32%, 22–49%, and 38–44%, respectively. The precipitation mode involves coprecipitation of multiple heavy metals to form a mineral. These heavy metals are adsorbed on the surface of bacteria through the participation of carboxyl, amino, and phosphate functional groups and extracellular polymeric substances. Therefore, S. marcescens C7-12 has strong biomineralization and adsorption capacity for heavy-metal ions in solution, which can provide potential resources for the bioremediation of heavy-metal-contaminated soil and water. Full article