Search Results (33,065)

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

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

Canine prostatic adenocarcinoma is a rare but highly aggressive cancer that is typically diagnosed at an advanced stage, due to the lack of effective screening methods and poor recognition of early lesions. Cancer stem cells are known to drive tumour progression and treatment resistance in human prostate cancer, but their role in naturally occurring canine disease remains poorly defined. A deeper understanding of the biology of canine prostatic adenocarcinoma is therefore essential to improve prognosis and to develop relevant comparative models. We established and comprehensively characterised two novel canine prostatic adenocarcinoma cell lines, Kodiak and Bobby, with detailed comparison to their tumours of origin and, for Kodiak, xenografts generated in immunodeficient mice. Both lines displayed variable epithelial morphology influenced by culture conditions, and Kodiak xenografts recapitulated key histopathological patterns of the primary tumour. Expression of the luminal epithelial marker CK8/18 and the basal marker CK14 was largely retained across tumour, cell line, and xenograft, whereas the basal markers CK5 and p63, and the urothelial marker UPIII, were diminished or lost during in vitro culture. Evaluation of cancer stem cell-associated markers showed consistent expression of CD44, Nanog, Oct3/4, and Sox2 in the original tumours and cell lines, while CD133, Nestin, and Trop2 were present in the tumours but absent in vitro, indicating selective loss of specific stem-like populations. Media-dependent plasticity was evident in the Bobby line. These models retain key epithelial and stemness features and provide robust platforms for translational prostate cancer research in dogs and humans. Full article

Molybdenum tellurite compounds have attracted increasing interest as promising nonlinear optical (NLO) materials, yet their microscopic second-harmonic generation (SHG) mechanisms remain unclear. In this work, the electronic structures and SHG responses of ATeMoO₆ (ATM, A = Mg, Cd, Zn) are systematically investigated using first-principles calculations combined with atom response theory. The results show that the SHG responses are mainly governed by the occupied nonbonding O 2p states and the unoccupied Mo 4d and Te 5p states. Our atom response theory analysis reveals that a strong synergistic effect between stereochemically active lone pairs (SCALPs) on Te atoms and nonbonding O 2p states critically enhances the SHG response in ZnTM and MgTM. In contrast, the relative weaker Te SCALPs in CdTM fail to provide a comparable contribution, leading to its lower SHG performance. The structure group analysis reveals that MoO₄ units dominate the SHG response, while TeO₄ units provide secondary contributions. Moreover, group dipole moments are found to be insufficient to explain the SHG behavior. These findings provide microscopic insights into SHG origins and offer guidance for NLO material design. Full article

Background and Objectives: Hypocalcemia due to hypoparathyroidism (HypoPTH) is the most common complication following thyroid surgery, typically resulting from iatrogenic removal, tissue damage, or compromised vascularization of the parathyroid glands. Patients with persistent HypoPTH are at risk for long-term complications such as osteoporosis, cardiac dysfunction, and renal impairment. Lifelong regulation of calcium levels is therefore essential to prevent morbidity and mortality associated with these complications. In this study, we aimed to evaluate the functional engraftment efficacy of 3D bioprinted human parathyroid tissue constructs in a xenograft model in vivo. Materials and Methods: Primary cells obtained from freshly excised human parathyroid tissue specimens were isolated and 3D bioprinted using alginate-based bioink. The bioprinted tissue constructs were implanted into CD1 athymic mice. Histopathological evaluation of the grafted constructs was performed at different time points. In addition, surface calcium-sensing receptor (CaSR) expression was assessed by immunofluorescence as an indicator of functional parathyroid tissue engraftment. Results: The presence of CaSR on parathyroid cells within the 3D-printed scaffolds confirmed the persistence of functional parathyroid cells following implantation. In tissue samples obtained during the first, second, and third weeks after implantation, CaSR positivity was consistently observed in the parathyroid cells. However, at the three-month follow-up, the pores within the scaffolds were found to be filled with calcified material and replaced by fibrotic tissue. At this stage, the absence of parathyroid hormone (PTH) expression indicated a loss of functional activity in the grafted biomaterial. Conclusions: Human primary parathyroid cells were successfully isolated, and a functional, hormone-active parathyroid tissue substitute was developed ex vivo using 3D-bioprinted hydrogel scaffolds combined with autologous cells. Although short-term functional engraftment was achieved, long-term graft viability and hormonal activity were limited due to scaffold degradation and fibrosis. These findings indicate the necessity for further improvement in scaffold biocompatibility to enhance the therapeutic potential of 3D-bioprinted parathyroid tissue constructs for in vivo applications. Full article

Extracellular vesicles, which carry bioactive cargos such as proteins, RNAs, and lipids, represent promising drug delivery vehicles owing to their biocompatibility, low immunogenicity, and inherent tissue-targeting capabilities. To address the current limitations in controlled cargo loading, we developed an abscisic acid (ABA)-inducible proximity system that directs proteins into exosomes during biogenesis. We engineered exosomal scaffolds by fusing the ABA receptor PYL1 to EV-enriched proteins—including BASP1, CD9, PTGFRN, and a truncated form PTGFRNΔ687—thereby creating docking sites within the exosomal lumen, while the target cargo (e.g., EGFP, firefly luciferase, or Cas9) was tagged with the ABI1 phosphatase domain. We demonstrate that ABA administration in producer cells induces PYL1–ABI1 complex formation, which recruits ABI1-fused cargo for selective encapsulation into EVs. Among the scaffolds tested, BASP1–PYL1 proved the most effective, enabling robust, ABA-dependent enrichment of cargo proteins. Purified EVs maintained canonical morphology, size, and marker expression (CD63, syntenin-1, CD9), confirming preserved biogenesis. Critically, these loaded exosomes efficiently delivered functional cargo to recipient cells, enabling Cas9/sgRNA-mediated genome editing. Together, our findings establish an ABA-triggered molecular switch for controllable EV protein loading, providing a versatile platform for next-generation therapeutic delivery. Full article

Urban storm drain sediments (SDSs) accumulate heavy metals from building façades and road surfaces, yet the biogeochemical controls governing metal mobility remain poorly understood. This study investigated biotic and abiotic controls on metal mobility along the urban dust transport chain (Xiamen-Quanzhou-Zhangzhou, China), using four sample types—façade dust (FD), road-deposited sediment (RDS), SDS, and runoff suspended solids (RSS)—from nine sites across three functional zones. Metal concentrations (Pb, Cu, Zn, Cr, Cd), phosphorus fractions, and microbial functional genes were quantified to test the hypothesis that viral abundance indicators, rather than pH, are more strongly associated with metal mobility in near-neutral urban sediments. Results showed that SDS served as metal accumulation hotspots with enrichment factors of 2.0–2.3× relative to FD, while total phosphorus declined by 34% along the transport chain. Contrary to conventional expectations, pH exhibited weak correlation with Pb mobility (r = −0.21; 95% CI: −0.62 to 0.27), whereas the T4-type bacteriophage gene g23 showed strong positive correlation (r = 0.85, p < 0.01; 95% CI: 0.52–0.96). Partial least squares path modeling revealed that viral abundance (g23 gene copies) showed the strongest statistical association with metal mobility among biotic variables (β = +0.48, p < 0.001), mediated through phosphorus-supported microbial activity. The model explained 76% of variance in metal mobility, with phosphorus cycling positively influencing viral abundance (β = +0.28). These findings challenge the pH-centric paradigm of metal geochemistry and reveal a novel phosphorus-virus-metal coupling mechanism in urban environments. The textile industrial site QZ-2 exceeded chromium screening values by 45%, demonstrating the framework’s utility for pollution hotspot identification. Full article

To ensure farmland soil quality and enable the systematic management of heavy metal contamination, this study assessed cadmium (Cd), mercury (Hg), and arsenic (As) levels in 114 surface soils from farmlands across four cities in Heilongjiang Province, China. The heavy metal elements were determined and comprehensively evaluated using four indices (geo-accumulation index, single factor contamination index, Nemerow index, and potential ecological risk index). Monitoring results showed that most soil samples exhibited concentrations of Cd, Hg and As that were below the risk screening values specified in relevant standards, except for those with slightly higher Cd concentration in a few soil samples from Suihua City. Comprehensive evaluation indicated a slightly to moderately Cd contamination across the study area, with Suihua City demonstrating the most prominent pollution levels. Factors such as railway traffic emissions might have caused Cd contamination in the surrounding farmland soil. Health risk assessments confirmed that the study area posed no significant carcinogenic or non-carcinogenic risks to children or adults via soil exposure. These findings highlight the need for priority monitoring of heavy metal concentrations and implementation of targeted soil protection measures to ensure both human health and sustainable agricultural development. It is necessary to closely monitor various potential pollution sources during the operation of surrounding railway lines, which is to prevent the formation of Cd-contaminated areas due to the long-term effects of railway transportation. The results of this study will have certain significance for the evaluation and management of regional farmland land pollution. Full article

Background: Indole-3-acetic acid (IAA)-based hydrazone derivatives, exemplified by specifically (E)-2-(1H-indol-3-yl)-N′-(3-methoxybenzylidene) acetohydrazide acetohydrazide (MBIH) and (E)-N′-(4-fluorobenzylidene)-2-(1H-indol-3-yl) acetohydrazide (FBIH), have garnered significant attention in the field of heavy metal toxicity for their potent antioxidant and cytoprotective properties. Methods: This study evaluated their efficacy, alongside ascorbic acid (AA), in mitigating sub-chronic cadmium (Cd) toxicity in a rat model. Sixty Swiss albino rats were randomized into five groups: control, Cd-exposed, Cd + AA (100 mg/kg), Cd + MBIH (10 mg/kg), and Cd + FBIH (10 mg/kg). Following 28 days of treatment, we assessed body weight trajectories, fasting blood glucose, and HbA1c. Serum biomarkers of hepatic, renal, inflammatory, and lipid function were quantified. Antioxidant capacity was measured via glutathione (GSH) assays and qRT-PCR analysis of SOD2, CAT, Nrf2, and Hmox 1 expression. Untargeted LC–MS/MS metabolomic profiling of serum identified disturbances in amino acids and lipid species, while histopathology of brain, liver, and pancreas documented structural injury. Results: Cd exposure induced significant weight loss, hyperglycemia, and elevated HbA1c, alongside dyslipidemia and heightened inflammatory markers. Hepatic and renal dysfunction, GSH depletion, and downregulation of antioxidant genes confirmed oxidative stress. Metabolomics revealed a Cd-specific fingerprint characterized by altered sulfur amino acid and phospholipid metabolism. Histologically, Cd caused liquefactive necrosis in the brain, inflammatory infiltrates in the liver, and acinar cell vacuolization with islet distortion in the pancreas. In contrast, MBIH and FBIH restored glycemic control, lipid profiles, inflammatory and hepatic renal markers, replenished GSH, and upregulated antioxidant genes via robust Nrf2 activation. Conclusions: These findings demonstrate that IAA-based hydrazone derivatives MBIH and FBIH afford superior protection against Cd-induced multi organ injury compared to ascorbic acid. Full article