Search Results (33,073)

Cyanobacteria of the genus Phormidesmis are recognized as a promising source of biologically active secondary metabolites with anticancer and immunomodulatory properties. In the present study, we investigated both the cytotoxic and immunological effects of an extract obtained from Phormidesmis molle PACC (Plovdiv Algal Culture Collection) 8140 as well as its chemical composition. The extract was profiled by LC-ESI-MS/MS (Liquid chromatography—electrospray ionization—tandem mass spectrometry), and selected compounds were evaluated with in silico ADMET (Absorption, distribution, metabolism, excretion and toxicity) modeling. The cytotoxic potential of the extract was evaluated in vitro using the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay on human colorectal adenocarcinoma cell lines (Caco-2, HT-29, and LS-180). The immunological impact of the extract was assessed on human peripheral blood mononuclear cells (PBMCs) isolated from healthy donors. PBMCs were treated with 100 µg/mL extract for 48 h, followed by flow cytometric immunophenotyping and ELISA (Enzyme-linked immunosorbent assay)-based cytokine quantification. The extract induced a concentration- and time-dependent decrease in cancer cell viability after 24, 48, and 72 h of exposure. At 72 h, treatment with the highest concentration (200 µg/mL) reduced cell viability to 74% in Caco-2 cells, 69–70% in HT-29 cells, and 59–61% in LS-180 cells. Morphological changes observed after treatment with Phormidesmis extract showed pronounced cytotoxic and apoptosis-related effects in the colorectal cancer cell lines tested. Immunophenotyping revealed a pronounced expansion of natural killer (NK) cells (CD56⁺ and/or CD16⁺). CD3⁻CD56⁻CD16⁺ NK population was markedly increased (from 67.7 ± 0.95% in non-treated PBMCs to 94.66 ± 0.90% in extract-treated PBMCs, p < 0.001). In contrast, the proportions of CD8⁺ T cells, CD19⁺ B cells, and CD11b⁺ monocytes were significantly reduced (from 21.5 ± 4.50% to 7.22 ± 0.41%, from 11.9 ± 1.70% to 6.06 ± 0.42%, and from 66.4 ± 0.60% to 34.4 ± 0.87%, respectively). Cytokine analysis demonstrated strong suppression of Th1-associated cytokines, with significantly reduced interferon gamma (IFN-γ, 461 ng/mL in controls vs. 84 ng/mL in extract-treated cultures) and tumor necrosis factor alpha (TNF-α) levels (169 ng/mL in controls vs. 32 ng/mL in extract-treated cultures), whereas nterleukin-6 (IL-6) was moderately elevated (from 158 ng/mL in controls to 234 ng/mL in extract-treated cultures) and IL-10 remained low. These findings demonstrate that P. molle extract combines cytotoxic activity against cancer cells with potent immunomodulatory effects, highlighting its potential as a source of bioactive compounds for immune-based therapeutic strategies. Full article

Background/Objectives: PAX6 haploinsufficiency-related congenital aniridia is a panocular disease affecting multiple ocular structures. The aim of this study was to determine the biometric properties of eyes affected by PAX6 haploinsufficiency-related classical congenital aniridia using a non-contact device. Methods: Fifty-nine eyes from 31 aniridia patients (48.39% male; mean age 27.0 ± 17.65 years, range 7–56) and 99 eyes from 50 healthy controls (44.00% male; mean age 28.56 ± 21.73 years, range 4–81) were examined using the Movu biometer (Argos Inc.). Axial length (AL), corneal diameter (CD), central corneal thickness (CCT), anterior chamber depth (ACD), lens thickness (LT), pupil size (PS), and mean keratometric value (K-mean) were measured. Results: Linear mixed-effects models showed significant effects of diagnosis on CCT (β = 182.39, p < 0.001), CD (β = −0.55, p = 0.02), and K-mean (β = −1.10, p = 0.03), while axial length was associated with gender (β = −0.90, p = 0.03). Mann–Whitney testing showed no interocular asymmetry (all p ≥ 0.07; η² ≤ 0.04) overall. Conclusions: PAX6-related congenital aniridia eyes are associated with increased CCT and reduced CD, and K-mean, while AL appears to be mainly influenced by gender. The absence of marked interocular asymmetry suggests relatively symmetrical bilateral involvement. These differences should be considered in corneal and lens surgery planning. Full article

Alveolar macrophages orchestrate phagocytosis and inflammatory programs during respiratory infection. CD2-associated protein (CD2AP) and BTB and CNC homology 2 (BACH2) are immune-related genes involved in cytoskeletal organization/vesicular trafficking and transcriptional regulation, respectively, but the coding-level constraints shaping their synonymous-site architecture remain unclear. Here, we profiled codon usage bias (CUB) of CD2AP and BACH2 across 49 vertebrate species using nucleotide composition, relative synonymous codon usage, and complementary codon bias diagnostics. Across species, BACH2 preferentially used G/C-ending codons with higher GC3s, whereas CD2AP was enriched for A/T-ending codons with lower GC3s. Both genes showed weak-to-moderate CUB (high ENC and modest CAI). ENC–GC3s and PR2 analyses indicated a strong compositional background at third codon positions, while neutrality analysis yielded shallow GC12–GC3 slopes, suggesting overall coding constraints, with compositional effects acting as a background influence and selective constraints possibly contributing to GC1/GC2 patterns. CD2AP deviated more from composition-only expectations than BACH2, suggesting greater gene-specific modulation. Phylogenetic reconstruction placed Sus scrofa within mammalian clades for both genes. In conclusion, CD2AP and BACH2 display opposite third-base codon-ending preferences across vertebrates, with CD2AP favoring A/T-ending codons and BACH2 favoring G/C-ending codons. This provides a comparative baseline for codon usage analyses of macrophage-relevant immune genes. Full article

Background and Clinical Significance: Plasmablastic lymphoma (PBL) is a rare and highly aggressive B-cell neoplasm most often associated with immunodeficiency. Transformation of chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL) into PBL is exceptionally uncommon, particularly in immunocompetent individuals. This paper describes a rare synchronous SLL-to-PBL transformation and summarizes current knowledge on synchronous and metachronous cases reported in the literature. Case Presentation A midle-aged immunocompetent patent presented with generalized lymphadenopathy and lumbar pain. Concurrent biopsies of an axillary lymph node and a retroperitoneal mass were obtained. Diagnostic evaluation included immunohistochemistry; fluorescent in situ hybridization (FISH); PCR-based assessment of IGH, IGK, and IGL loci; and next-generation sequencing (NGS) of IGHV to assess clonal relatedness. The patient was treated with six cycles of Dara-CHOP, followed by autologous stem cell transplantation and maintenance therapy with daratumumab and ibrutinib. The axillary node showed SLL (CD20+, CD5+, CD23+), while the retroperitoneal mass demonstrated classic features of PBL (CD138+, MUM1+, MYC+, Ki-67 ~100%, CD20−). FISH detected MYC rearrangement in the PBL component. PCR and NGS confirmed identical IGHV1-69 rearrangements, establishing clonal relatedness and Richter transformation. A review of published cases shows that both synchronous and metachronous CLL/SLL-to-PBL transformations are exceedingly rare. The patient achieved partial metabolic remission after treatment and remains in sustained metabolic response 24 months after diagnosis. Conclusions: This case highlights a rare example of synchronous CLL/SLL-to-PBL transformation in an immunocompetent patient. Integration of detailed molecular diagnostics enabled early recognition and guided a personalized treatment approach incorporating CD38-targeted therapy and BTK inhibition, resulting in an excellent long-term clinical outcome. Full article

Background/Objectives: The ketogenic diet (KD) has been shown to exert beneficial effects on human immunity by enhancing cytotoxic T lymphocyte function through metabolic reprogramming. However, strict dietary restrictions limit adherence and complicate its use in clinical practice. Exogenous ketone supplements have therefore been promoted as a more feasible alternative to elevate ketone body levels without the need for dietary changes. The objective of this study was to assess whether ketone salt or ketone ester supplementation can reproduce KD-mediated immunometabolic effects on CD8⁺ T cells in healthy individuals. Methods: In a prospective interventional study, healthy volunteers received either ketone salts (KS) or ketone esters (KE) for three weeks. Plasma β-hydroxybutyrate (BHB) concentrations were determined, and CD8⁺ T-cell cytokine secretion, functional responses, and mitochondrial energy metabolism were analyzed. In a subgroup, KS supplementation was combined with a carbohydrate-restricted, non-ketogenic diet. Results: While KS supplementation resulted in a short-lived increase in plasma BHB concentrations followed by increased BHB uptake in immune cells, KE supplementation led to more sustained plasma BHB levels, however, without detectable intracellular BHB accumulation. Neither intervention affected CD8⁺ T-cell cytokine production, functional capacity, or mitochondrial energy metabolism, and KS intake combined with a carbohydrate-restricted, non-ketogenic diet likewise did not alter CD8⁺ T-cell immunometabolic parameters. Conclusions: Transient elevation of circulating ketone body levels through supplementation seems insufficient to reproduce the immunometabolic effects of a KD, which likely require broader metabolic adaptations. Thus, the impact of exogenous ketones on adaptive immunity in healthy individuals appears limited. Full article

Geranium robertianum L. is used in traditional medicine to treat different systemic disorders and holds great therapeutic potential but remains understudied. To this aim, an ethanolic extract obtained from the aerial parts of G. robertianum L. (GR) was investigated in terms of phytochemical composition and biological activity. GR extract exhibited high levels of phenolic compounds and flavonoids. The antioxidant activity was determined by means of three different colorimetric assays (DPPH, ABTS, and FRAP), and the results obtained indicate that the ABTS assay showed the highest antioxidant capacity. Metal analysis was also performed. Fe was found to be the most abundant element in the analyzed extract, with a concentration of 363.65 ± 4.18 μg/g, followed by Zn, Mn, Ni, and Cr. Four potentially hazardous heavy metals, As, Co, Pb, and Cd, were found to be under the detection limit. The GR extract exhibited moderate antibacterial activity against both Gram-positive and Gram-negative bacteria, with inhibition zones generally comparable to those of levofloxacin. However, the extract was significantly less effective against the P. aeruginosa strain. On A253 human salivary gland carcinoma cells, GR extract elicited a dose-dependent antiproliferative effect, produced morphological changes, and increased ROS and both caspase-3/7 and caspase-9 levels. Full article

This study comprehensively evaluates the elemental composition of soil and Pinus species needle samples across 25 distinct plots established along the D825 highway in Kahramanmaraş, Türkiye. Located at the confluence of the Mediterranean, East Anatolian, and Central Anatolian regions, this area represents a critical ecological transition zone. A total of 75 soil and 75 needle samples were analyzed in triplicate to assess heavy metal contamination and potential toxicity risks across these elevation gradients. According to the results, the Geoaccumulation Index (I_geo) values for all examined metals remained below zero, categorizing the study area as “unpolluted.” Enrichment Factor (EF) analyses confirmed the lithogenic origin of Cr, Mn, and Ni; however, Lead (Pb) and Cadmium (Cd) exhibited an EF of 1.34. This ‘minimal enrichment’ could potentially be associated with anthropogenic pressures, possibly stemming from traffic emissions on the highway. Although current metal levels fall below global toxicity thresholds (WHO/FAO), the positive skewness and high variation in Pb and Cd distributions suggest a likelihood of localized accumulation, which may warrant systematic monitoring. The original contribution of this study lies in its integrated assessment of plant–soil barrier mechanisms within this unique transition zone, demonstrating how forest ecosystems maintain resilience capacity despite ophiolitic parent material contributions. While soil Cr and Ni levels were elevated due to the geological structure, plant tissue concentrations remained within safe physiological limits, suggesting effective stabilization within the soil-biomass matrix. The findings suggest that these forest ecosystems play a key role in maintaining ecological health and environmental sustainability against potential anthropogenic encroachment in this strategic intersection. Full article

Artificial intelligence (AI) is a cutting-edge technology that can replicate knowledge, operation and, at some point, understanding at a human-like level. The AWARE project aims to develop an AI assistant application (ASA) designed to support air traffic control (ATC) operations by building a platform based on enhanced artificial situational awareness. One of the pillars of the ASA system is to develop a set of functionalities that mimic the behavior of human actions based on the development of technical tools. Regarding safety issues, conflict detection and resolution (CD&R) is the pillar to identify conflicts and avoid mid-air collisions. The goal is to build a CD&R that can be embedded into the ASA system and generate outputs that can be usable and valuable for ATC. CD&R tool is based on two subsystems: The CD component identifies potential separation minima infringements, while the CR module produces explainable resolution maneuvers with standardized syntax for seamless ATCO integration. CD uses a deterministic algorithmic approach grounded in trajectory prediction models, while CR implements a hierarchical decision-making architecture that emulates expert ATCO cognitive processes within a client-service paradigm where pilots serve as end-users. Full article

Anti-β2GPI/HLA-DR antibody, chronic endometritis (CE), and endometrial dysbiosis are likely to be associated with the etiologies of recurrent pregnancy loss (RPL). This prospective cohort study aimed to investigate these new risk factors together with conventional causes for RPL, and to evaluate pregnancy outcomes in women individually treated. A total of 87 women with RPL underwent conventional assessment together with anti-β2GPI/HLA-DR antibody measurements, CD138 immunohistochemistry for CE, and 16S rRNA sequence analysis for endometrial microbiome. Women with anti-β2GPI/HLA-DR antibody, CE, and endometrial dysbiosis received low-dose aspirin and heparin, antibiotics, and probiotics, respectively. Pregnancy outcomes of the participants were assessed. Anti-β2GPI/HLA-DR antibody, CE, non-Lactobacillus-dominant microbiome (NLDM)-1 (Lactobacillus + Bifidobacterium < 80%), and NLDM-2 (Lactobacillus without iners + Bifidobacterium < 80%) were detected in 16 (18.4%), 22 (25.3%), 27 (31.0%), and 46 (52.8%) women, respectively. Based on conventional assessment, 65.5% of women with RPL were classified as unexplained etiology; however, the percentage reduced to 16.1% when these new tests were assessed together. All 9 pregnancies with anti-β2GPI/HLA-DR antibody, 13 (92.9%) of 14 pregnancies with CE, and 24 (92.3%) of 26 pregnancies with NLDM-2 resulted in live birth. Assessment of these new tests may be clinically useful for reducing the proportion of unexplained RPL, and for providing high live birth rates if women receive relevant treatments. Full article

Pancreatic ductal adenocarcinoma (PDAC) is characterized by immune cell dysfunction and poor prognosis. CD44, a cell surface glycoprotein with multiple splice variants, has been implicated in tumor progression, but its compartment-specific roles in PDAC remain unclear. CD44 standard and variant isoform expression was analyzed in patient-derived lymph nodes (LNs) by quantitative PCR. Immune cell-specific CD44 expression was assessed by flow cytometry in LNs and peripheral blood. Soluble and exosome-associated CD44 (exo-CD44) were measured in plasma. Clinical associations and survival analyses were performed. Transcriptomic, immune infiltration, immune checkpoint, and drug sensitivity analyses were conducted using TCGA-PAAD and pharmacogenomic datasets. CD44 standard isoform expression was unchanged in PDAC LNs, whereas multiple CD44 variant isoforms (v4–v10) were significantly reduced and associated with metastatic disease and poor survival, particularly CD44v5, v6, v7, and v10. CD44 expression was enriched in CD45⁺ immune cells, with highest levels in CD4⁺ T cells in both LNs and blood. Soluble CD44 levels showed no clinical associations. In contrast, exo-CD44 levels were reduced overall in PDAC but increased in patients with distant metastasis, positive resection margins, systemic inflammation, and reduced survival. High CD44 expression was associated with advanced disease, immune cell infiltration, immune checkpoint gene expression, reduced sensitivity to gemcitabine, paclitaxel, rapamycin, and FMK, and distinct CTLA4/PD-L1 checkpoint profiles. CD44 exhibits compartment-specific regulation in PDAC, linking immune remodeling, exosome signaling, and therapeutic resistance to adverse clinical outcome. Full article

Cephalopods and crustaceans are known to bioaccumulate heavy metals, potentially posing both non-carcinogenic and carcinogenic health risks to consumers. This study was conducted to determine heavy metal concentrations and assess associated health risks in the edible tissues of 84 cephalopod and crustacean samples. Heavy metal concentrations and assess associated health risks in the edible tissues of 84 cephalopod and crustacean samples collected from selected wholesale markets and major fish landing ports throughout Peninsular Malaysia. The analysis focused on nine heavy metals: selenium (Se), cadmium (Cd), lead (Pb), copper (Cu), zinc (Zn), antimony (Sb), tin (Sn), chromium (Cr), and manganese (Mn). The samples were digested using a microwave digestion system, and heavy metal concentrations were analysed using Inductively Coupled Plasma Mass Spectrometry (ICP-MS). Results showed that Mn was the most abundant metal, followed by Cr and Zn. Octopus (C. indicus) had the highest Mn concentration (5.01 mg/kg WW), while Rainbow shrimp (P. sculptilis) had the highest overall metal concentration (91.02 mg/kg WW). Significant differences were observed between cephalopods and crustaceans, with Cd and Sn concentrations being notably higher in cephalopods (p < 0.001). However, no significant associations were observed between heavy metal concentrations and sample weight or length, indicating a greater influence of environmental factors. Principal Component Analysis (PCA) explained 80.4% of the variance, with Cd, Sn, Pb, Cu, Zn, Cr, and Mn accounting for the majority of the variance. Estimated weekly intake (EWI) values ranged from 0.002 to 26.30 µg/kg bw/week for cephalopods and 8.02 × 10⁻⁶ to 243.175 µg/kg bw/week for crustaceans. All metal levels were below the permissible limits set by the Food and Agriculture Organisation of the United Nations/World Health Organisation (FAO/WHO). Hazard Index values were <1, indicating low non-carcinogenic risk, and Total Carcinogenic Risk values for Pb and Cr were below 1 × 10⁻⁴, suggesting negligible carcinogenic risk. Full article

Osteal macrophages (“osteomacs”) are resident bone macrophages that support osteoblast differentiation and bone formation. Despite their importance in bone homeostasis, their function in human bone metabolism and osteoporosis remains poorly understood, largely due to the lack of a standardized isolation protocol. Here, we present a protocol for isolating primary human osteomacs from femoral head specimens obtained during arthroplasty. After the removal of bone marrow to minimize contamination with marrow-derived macrophages, bone fragments were enzymatically digested and osteomacs were isolated using CD14-based MACS^® or CD14/CD45/ALP-based FACS. Immunofluorescence confirmed macrophage identity and revealed expression of markers associated with both M1-like and M2-like activation states. Isolated cells displayed heterogeneous morphology and could be maintained in culture. This protocol enables reproducible isolation of human osteomacs and provides a foundation for translational studies investigating osteoimmune interactions in bone disease and osteoporosis. Full article

Background/Objectives: Tuberculosis (TB) remains a major global health challenge, and the Bacillus Calmette–Guérin (BCG) vaccine has limited efficacy against adult pulmonary disease. Protein subunit vaccines are a promising alternative but require strong adjuvants to induce cell-mediated immunity. Synthetic agonists targeting toll-like receptor 4 (TLR4) and stimulators of interferon genes (STINGs) have emerged as effective immunostimulants. Therefore, we aimed to evaluate the immunogenicity and protective efficacy of Ag85B-based subunit vaccines formulated with synthetic TLR4 and STING agonists in a BCG-boosted mouse model. Methods: Three synthetic adjuvants—QTP709-1, QTP709-3, and QTP701—were formulated as oil-in-water emulsions containing distinct surfactant and immunostimulant components. The potential of vaccine formulations to activate dendritic cells (DCs) and elicit Ag85B-specific immune responses, including IgG subclass levels, interferon-γ (IFN-γ) enzyme-linked immunosorbent spots, and polyfunctional T-cell responses, was assessed by flow cytometry. Protective efficacy was evaluated based on pulmonary bacterial burden and histopathology following Mycobacterium tuberculosis (M. tb) Erdman challenge. Results: All formulations promoted DC maturation and enhanced antigen-specific immune responses. Each adjuvant elicited strong Ag85B-specific humoral immunity, increased IFN-γ secretion, and polyfunctional CD4⁺ and CD8⁺ T cells co-producing IFN-γ, TNF-α, and interleukin-2. Among them, QTP709-1 was associated with increased levels of chemokine receptor 5-associated chemokines and showed a trend toward reduced lung bacterial burden and histopathological inflammation following M. tb challenge. Conclusions: Synthetic TLR4 and STING agonists were associated with enhanced immunogenicity of TB subunit vaccines and showed evidence of protective potential, with TLR4-based formulations exhibiting more pronounced immunological responses. QTP709-1 exhibited strong immunostimulatory and protective effects, supporting its potential as a candidate adjuvant for next-generation TB vaccines. Full article

Late Permian paleoenvironmental instability and recurrent biotic crises coincided with enhanced marine organic-carbon burial, yet ocean-circulation dynamics have remained underappreciated as a key driver. In particular, for the Wuchiaping Formation along the eastern margin of the Paleo-Tethys Ocean, the presence, variability, and mechanistic impact of upwelling—and its coupling with water-column redox structures—have not been systematically constrained, limiting a process-based understanding of organic-matter enrichment. Here, we integrate sedimentological, mineralogical, and multi-proxy geochemical data to investigate the dominant controls on organic matter enrichment in the Wuchiaping Formation shale succession from the northeastern Sichuan Basin. The Lower Wuchiaping Formation consists mainly of clay-rich shales deposited under oxic, shallow-water, and weakly stratified conditions, as indicated by low Ni/Co ratios (average 1.88), limited uranium enrichment (U_EF = 0.21), low Ba/Al ratios, and sparse biogenic debris. Biomarker indices (gammacerane index = 0.35; Pr/Ph = 1.91) suggest unfavorable preservation conditions, resulting in a low mean TOC of 0.78%. In contrast, the Upper Wuchiaping Formation is dominated by siliceous shales with elevated Ni/Co ratios (average 15.83), moderate uranium enrichment (U_EF = 2.48), abundant framboidal pyrite, radiolarian–planktic foraminiferal assemblages, and laminated apatite. High Ba/Al and Cd/Mo ratios, higher gammacerane values, and low Pr/Ph ratios (<1) indicate enhanced water-column stratification and bottom-water anoxia, leading to efficient organic matter preservation and a high mean TOC of 9.2%. Biomarker compositions reveal a shift from terrestrial-dominated organic matter in the Lower Wuchiaping Formation to algal- and plankton-derived inputs in the Upper Wuchiaping Formation. Collectively, these results indicate that intensified upwelling—rather than tectono-magmatic forcing alone—was the primary driver of enhanced productivity, strengthened redox stratification, and organic matter enrichment in the Upper Wuchiaping Formation. Our findings highlight the importance of upwelling–redox coupling as a key mechanism linking Late Permian ocean-system reorganization to spatially and stratigraphically heterogeneous organic-carbon accumulation along the Paleo-Tethyan margin. Full article

Crohn’s disease (CD) is characterized by chronic intestinal inflammation accompanied by gut dysbiosis and redox imbalance. We investigated the role of dual oxidase-2 (DUOX2), a major epithelial source of reactive oxygen species (ROS), in linking oxidative stress to microbe–host crosstalk. DUOX2 expression was upregulated in human intestinal samples and was positively associated with inflammatory readouts, oxidative stress indices, and dysbiosis. Intestinal epithelial cell-specific Duox2 knockout (KO) mice exhibited reduced mucosal ROS, preserved barrier integrity, and attenuated dextran sodium sulfate (DSS)- and 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced colitis. Cohousing and fecal microbiota transplantation demonstrated that this protective phenotype was microbiota-dependent. Multi-omics profiling identified enrichment of Parabacteroides, particularly P. distasonis, in Duox2 KO mice, and oral supplementation with P. distasonis enhanced resistance to colitis. Mechanistically, DUOX2-derived oxidative stress constrained Parabacteroides growth, as P. distasonis displayed marked susceptibility to hydrogen peroxide, with excessive intracellular ROS accumulation and an absence of key antioxidant defenses—including peroxide reductase C (AhpC) and superoxide dismutase B (SodB)—indicating that epithelial DUOX2 shapes a hostile luminal redox niche unfavorable to these beneficial microbes. Pharmacological inhibition of DUOX2 with Compound 521 reduced oxidative stress, ameliorated colitis, and partially restored microbial balance. These findings establish a DUOX2–ROS–microbiota axis in which epithelial DUOX2 amplifies oxidative stress, remodels the gut ecosystem, and promotes inflammation, and highlights DUOX2 suppression or ROS-sensitive Parabacteroides as potential redox-centric therapeutic strategies for CD. Full article