Search Results (50,422)

Background: Hashimoto’s thyroiditis (HT) is a chronic autoimmune disorder characterized not only by thyroid dysfunction but also by metabolic disturbances, micronutrient inadequacies, and low-grade inflammation. Composite indices derived from routine laboratory parameters may therefore help capture the broader systemic profile of the disease. This study explored within-cohort associations of immunonutritional indices including the Prognostic Nutritional Index (PNI), Nutritional Risk Index (NRI), and Controlling Nutritional Status (CONUT), and hemogram-derived inflammatory markers including the Neutrophil-to-Lymphocyte Ratio (NLR), Monocyte-to-Lymphocyte Ratio (MLR), Platelet-to-Lymphocyte Ratio (PLR), and Systemic Immune-Inflammation Index (SII), with thyroid function, thyroid autoimmunity, metabolic characteristics, disease duration, and vitamin D status in adults with Hashimoto’s thyroiditis. Methods: This cross-sectional study included 229 adults diagnosed with HT. PNI, NRI, CONUT, and complete blood count-derived inflammatory markers were evaluated in relation to thyroid function, thyroid autoimmunity, disease duration, metabolic characteristics, and vitamin D status. Because most variables were not normally distributed, the main analyses were conducted using non-parametric tests. Correlations were evaluated using Spearman’s rank correlation coefficients. Exploratory regression models were estimated using HC3 heteroscedasticity-consistent robust standard errors, and CRP-based sensitivity analyses were performed by excluding participants with CRP > 10 mg/L. Results: Vitamin D deficiency was highly prevalent and affected 70.3% of the participants. Among the immunonutritional indices, NRI differed significantly according to BMI category and HOMA-defined insulin resistance (both p < 0.001), indicating a closer relationship with metabolic burden. PNI was associated with disease duration (p = 0.009), whereas the inflammatory indices were largely similar across the clinical groupings examined. In exploratory robust regression models, the explanatory power remained modest (R² = 0.066–0.171). PLR showed the most consistent index-related association with TSH, whereas the CONUT–FT3 association observed in the full-sample robust model was not retained after CRP-based sensitivity analysis. Conclusions: Adults with HT in this study showed frequent vitamin D deficiency together with a substantial burden of excess weight and insulin resistance. Routine immunonutritional and inflammatory indices may provide supportive information on within-cohort biochemical and metabolic heterogeneity, but they should not be interpreted as stand-alone diagnostic or prognostic markers. In particular, NRI appeared to reflect metabolic and adiposity-related burden more than nutritional risk alone, while PLR showed the most internally consistent index-related association with TSH. Full article

Although Tenebrio molitor is increasingly explored as a feed ingredient, uncertainty remains regarding the effects of high dietary inclusion levels in model animals. Therefore, this study evaluated physiological and behavioral responses to graded inclusion levels of T. molitor meal (25–45%) in the diets of adult female Wistar rats. The assessed outcomes comprised (i) clinical parameters, (ii) food and water intake, (iii) hematology and serum biochemistry, and (iv) behavior. Across the insect-fed groups, no unintended weight loss or excessive gain was observed; instead, animals modulated their feed intake in a manner consistent with maintaining a relatively stable growth trajectory, without signs of hyperphagia suggestive of nutrient deficiency or anorexia indicative of adverse effects under study conditions. Diet acceptance remained high throughout the experiment, with no feed refusal or behavioral evidence of aversion that might be expected in response to poor palatability or discomfort. Heart rates and respiratory rates remained age-appropriate, showing typical temporal variability. Significant between-group differences were observed in RDW, Retic (%), NEU (%, K/μL), EOS, PLT, and PCT (p < 0.05), while serum analyses revealed significant differences in AST, total bilirubin, creatinine, phosphorus, and BUN (p < 0.05). A moderate negative correlation between individual feed intake and body weight was additionally identified in selected groups. The results of behavioral testing reveal that the dietary inclusion of T. molitor meal at levels up to 45% did not reduce spontaneous activity or elicit anxiety-like responses, and exploratory behavior appeared to be preserved. Overall, the present findings indicate that the gradual incorporation of T. molitor meal at high inclusion levels supported normal growth, physiological function, and behavior in rats under the conditions studied. Full article

The Ros/MucR family is constituted by proteins controlling the expression of genes crucial for the interaction with eukaryotic hosts. Ros/MucR family members were classified as H-NS-like proteins in α-proteobacteria, as they share fundamental features with H-NS proteins playing a pivotal role in controlling gene expression by structuring the bacterial genome. Here, we identified two new Ros/MucR family members in Sinorhizobium meliloti. They differ from classical MucR homologs since MucR2 lacks the circular oligomeric structure typical of other family members and MucR3 shows a concentration-dependent oligomerization ability with a low propensity to form circular particles, as shown by cryogenic electron microscopy. Moreover, MucR2 and MucR3 present a new zinc coordination sphere. The newly identified MucRs bind DNA, but lack the DNA bridging activity, which is crucial for structuring the bacterial genome. Using mass spectrometry, light scattering, NMR, EMSA and bridging assay, our study reports the first identification and characterization of two new MucRs and indicates that Ros/MucR family members control gene expression through distinct mechanisms. These results provide an important framework for future studies aimed at dissecting the interplay among MucR proteins and understanding how they can jointly orchestrate condition-dependent gene expression in bacterial species expressing multiple mucR homologous genes. Full article

Hierarchical functionalisation of the UiO-66(Zr)-NH₂ metal–organic framework with cysteine, poly(ethylene glycol) (PEG), and the SARS-CoV-2 spike receptor-binding domain (RBD) was developed to enable receptor-specific interaction with the angiotensin-converting enzyme 2 receptor (ACE2) in model cells. Post-synthetic modification using cysteine and heterobifunctional PEG linkers allowed controlled bioconjugation of SpyTag-labelled RBD via SpyTag/SpyCatcher chemistry, while preserving the crystallinity, microporosity, and intrinsic optical properties of the UiO-66(Zr)-NH₂ framework. Comprehensive physicochemical characterisation confirmed successful surface functionalisation, tunable aggregation behaviour, and retention of multimodal optical characteristics. Cellular studies in HEK293T and HeLa cells overexpressing EGFP-tagged ACE2 demonstrated enhanced and selective association and uptake of RBD-functionalised nanoparticles compared with non-targeted analogues. Multimodal fluorescence imaging, fluorescence lifetime imaging microscopy, flow-cytometry, and electron microscopy indicated ACE2-dependent endocytic internalisation, with predominant localisation in endosomal and autophagosomal compartments, while both amine- and cysteine-modified formulations exhibited good biocompatibility. Overall, this study establishes a virus-mimetic, ACE2-targeted UiO-66(Zr)-based nanosystem as a proof-of-concept biointerface platform for receptor-specific cellular delivery and imaging, providing a foundation for future MOF-based nanocarriers exploiting ligand–receptor interactions. Full article

Background/Objectives: Spinal muscular atrophy (SMA) is a rare genetic disorder caused by mutations or deletions in SMN1, resulting in the loss of SMN protein and severe neuromuscular consequences. Nusinersen, an antisense oligonucleotide that promotes full-length SMN2 transcript formation, has significantly improved SMA outcomes. However, standardized in vitro procedures for evaluating nusinersen efficacy remain limited. This study aimed to optimize in vitro efficacy assessment of nusinersen across two human cellular models. Methods: Experiments were performed using HEK293 cells and the SMA patient-derived fibroblast line GM03813. Transfection conditions were optimized for each model. In HEK293 cells, several seeding densities were evaluated for nucleofection, while in GM03813 fibroblasts, multiple transfection reagents and protocols were tested. Nusinersen activity was quantified at the transcript and protein levels, and dose–response curves were generated to determine EC₅₀ values. Results: In HEK293 cells, a higher seeding density (1 × 10⁶ cells) yielded the most efficient nucleofection. In GM03813 fibroblasts, Lipofectamine 3000 outperformed the other transfection reagents tested. Nusinersen exhibited dose-dependent effects in both models. The EC₅₀ for transcript induction in HEK293 cells was 293 nM, whereas in GM03813 fibroblasts the EC₅₀ was 10 nM, demonstrating substantial model-dependent differences in response. Conclusions: This study establishes optimized conditions for in vitro efficacy assessment of nusinersen in HEK293 and GM03813 cellular models. These protocols provide a robust and reproducible framework for evaluating nusinersen and can be readily applied to other antisense oligonucleotides designed to correct SMN2 splicing. Full article

Radiotherapy is a highly effective, safe cancer treatment, and about half of all cancer treatments involve lifesaving radiotherapy. Despite huge advances in technology that have made it safer and more effective, it is still not without side effects. They differ from patient to patient and can include fatigue, nausea, skin reactions, and hair loss, but dysbiosis is the most common complication associated with radiotherapy. Probiotics aimed at restoring the microbiome have found widespread use, but the problem of their rapid inactivation in the gastrointestinal tract has not yet been solved. Our study aims to confirm the effectiveness of a novel biomineral complex, based on a powdered clinoptilolite containing a rock loaded with lactobacilli for restoring the intestinal microbiome of mice exposed to radiation. Based on the 16S rRNA gene analysis, alpha-diversity and dynamics of changes in the fecal metagenome, as well as the functional potential of mice exposed to radiation, were studied, and the prospects of administering the biomineral complex to achieve positive effects were assessed. NMR analysis of the mineral carrier was carried out, and its safety was confirmed. Moreover, per os administration of the complex following irradiation led to a reduction in the level of chromosomal aberrations induced by irradiation. Thus, the biomineral complex has a microbiome-restoring effect and reduces radiation-induced clastogenesis. Full article

Background: Autism spectrum disorder (ASD) arises from complex gene–environment interactions. While trace element abnormalities have been studied, associations with autism severity remain inconsistent. Ratios indicating detoxification balance, rather than single toxic elements, may better reflect severity. Objective: To examine the relationships between urinary trace element levels, detoxification-related element ratios, and autism severity measured by the Childhood Autism Rating Scale (CARS). Methods: In a cross-sectional study of 168 participants (103 ASD, 65 controls), thirty urinary trace elements were quantified by ICP-MS. ASD patients were stratified by CARS into subthreshold ASD (n = 29), mild–moderate ASD (n = 36), and severe ASD (n = 38). Analyses included Mann–Whitney U, Kruskal–Wallis, and Spearman correlation tests, focusing on Li/Pb, Cu/Pb, and Cr/Pb ratios. Results: Individual elements showed weak associations with CARS; lead correlated positively (ρ = 0.209, p = 0.035) and lithium inversely (ρ = −0.194, p = 0.051). In contrast, element ratios showed stronger links: Li/Pb (ρ = −0.349, p = 0.0003), Cu/Pb (ρ = −0.320, p = 0.0011), and Cr/Pb (ρ = −0.209, p = 0.035). Severe ASD exhibited modest 90th-percentile elevations for toxic elements but high heterogeneity. Conclusions: Single-element levels showed limited associations with ASD severity. Element ratios, particularly Li/Pb, showed stronger statistical associations than individual elements in this cross-sectional dataset; however, these findings should be interpreted as candidate correlates rather than causal or clinically validated biomarkers. Full article

Background/Objectives: Sepsis is a life-threatening syndrome arising from a dysregulated host response to infection, frequently leading to multiple organ dysfunction, with the lungs being among the most severely affected organs. Oxidative stress, inflammation, apoptosis, and DNA damage play key roles in the pathogenesis of sepsis-induced acute lung injury (ALI). Beyond its lipid-lowering effects, rosuvastatin possesses anti-inflammatory and antioxidant properties that may confer protective effects in sepsis. This study was designed to investigate the dose-dependent prophylactic efficacy of rosuvastatin in mitigating pulmonary damage in rats with cecal ligation and puncture (CLP)-induced sepsis. Methods: Sprague–Dawley rats were randomly divided into six groups: Sham, Sham + rosuvastatin (10 mg/kg), Sham + rosuvastatin (20 mg/kg), CLP, CLP + rosuvastatin (10 mg/kg), and CLP + rosuvastatin (20 mg/kg). Rosuvastatin was administered via oral gavage 4 h before the surgical procedures in the experimental groups. All animals were sacrificed 16 h following surgical procedures. Lung tissues were analyzed for biochemical markers, including malondialdehyde (MDA) and reduced glutathione (GSH), as well as histopathological changes and immunohistochemical expression of NF-κB/p65, caspase-3, and 8-OHdG. Results: CLP-induced sepsis significantly increased MDA levels while decreasing GSH levels, indicating enhanced oxidative stress. Rosuvastatin treatment significantly reversed these changes. Histopathological analysis revealed marked lung injury in the CLP group, including alveolar inflammation, interstitial inflammation, vascular congestion, and increased alveolar septal thickness, all of which were significantly reduced following rosuvastatin administration. Immunohistochemical findings demonstrated increased expression of NF-κB/p65, caspase-3, and 8-OHdG in the CLP group, whereas rosuvastatin significantly attenuated these expressions. No significant difference in prophylactic efficacy was observed between the 10 mg/kg and 20 mg/kg doses of rosuvastatin. Conclusions: Rosuvastatin demonstrated a protective effect against sepsis-induced pulmonary damage by reducing oxidative stress, inflammation, apoptosis, and DNA damage. These findings suggest that rosuvastatin may have prophylactic potential in sepsis; however, further support is needed from investigations of cellular pathways in different mechanistic directions. Full article

Several bioactive compounds, including phenolic and flavonoid substances, have been identified in the aqueous leaf extract of Schinus terebinthifolius (ALE). These compounds are active ingredients in green nanoparticle biosynthesis. Transmission electron microscopy (TEM), energy dispersive X-ray spectroscopy (EDX), zeta potential analysis, and FTIR spectral analysis were used to characterize copper oxide nanoparticles (CuNPs) and silver nanoparticles (AgNPs). According to TEM results, AgNPs exhibited somewhat larger diameters (12 ± 4 nm), were spherical with significant aggregation, and displayed a fairly uniform distribution, while CuNPs were primarily quasi-spherical with a narrow size range of about 4–5 nm. CuNPs showed a much more negative zeta potential value of −25.8 mV, indicating good to high colloidal stability, whereas AgNPs had a zeta potential of −15.5 mV, suggesting moderate stability. The main compounds included chlorogenic acid (10,375.28 µg/g), gallic acid (7015.59 µg/g extract), ellagic acid (1571.29 µg/g extract), and rutin (1485 µg/g extract). The antifungal activity of CuNPs and AgNPs was tested at concentrations of 6, 12, 25, 50, and 75 μg/mL on Quercus rubra wood against Fusarium circinatum and Pythium tardicrescens. The greatest inhibition of F. circinatum growth was observed with CuNPs and AgNPs at 75 µg/mL, showing fungal inhibition percentages (FIPs) of 61.48 and 60.74%, respectively. CuNPs and AgNPs at 75 µg/mL exhibited moderate activity against P. tardicrescens, with FIPs of 21.48% and 15.92%, respectively. The MICs for AgNPs and CuNPs were 1.5 and 85 µg/mL with F. circinatum and P. tardicrescens, respectively. Overall, CuNPs and AgNPs demonstrated potential antifungal activity against F. circinatum but moderate activity against P. tardicrescens compared to the control. This ALE from S. terebinthifolius is rich in flavonoids and phenolic compounds, including gallic acid, chlorogenic acid, rutin, ellagic acid, and p-coumaric acid, as identified by HPLC analysis. These biomolecules act as both capping agents, which stabilize the nanoparticles, and reducing agents. Using S. terebinthifolius ALE’s rich phytochemical profile as a reducing and stabilizing agent provides an environmentally friendly method for the green synthesis of CuNPs and AgNPs. Full article

This review examines Bacillus thuringiensis subsp. israelensis (Bti) as both a highly selective microbial larvicide and a biological platform for protein storage and delivery, enabled by the structural features of its prokaryotic insect larvicidal organelle (PILO). Bti remains the most widely deployed biological agent for mosquito control. Decades of operational use demonstrate substantial public health benefits and only limited, manageable ecological tradeoffs within integrated vector management programs (IVMP). Its narrow host range underlies an excellent safety record for humans and other vertebrates. Moreover, laboratory and field studies consistently show that collateral effects are minimal, context dependent, reversible, and largely restricted to closely related non-target aquatic dipterans. These attributes have established Bti as a cornerstone of environmentally sustainable IVMP worldwide. Here, we synthesize current knowledge on Bti biology, ecological selectivity, field performance, and the resistance-management properties embedded in the molecular architecture of the PILO. Finally, we assess emerging opportunities and technical constraints in repurposing the PILO as an in vivo microbial factory for packaging heterologous proteins with potential pharmaceutical and industrial applications. Full article

The role of human leukocyte antigen F (HLA-F) at the maternal–fetal interface (MFI) during viral infection and its regulation by interferon signaling remains poorly understood. Here, we investigated HLA-F expression and regulation in first-trimester trophoblast cells following activation of the type I interferon pathway and viral infection. We demonstrate that HLA-F is significantly upregulated at both mRNA and protein levels in response to Poly(I:C) and IFN-β in a dose- and time-dependent manner, suggesting its regulation as an interferon-stimulated gene (ISG). Zika virus (ZIKV) infection similarly induced HLA-F upregulation over time. In contrast, HSV-2 infection downregulated HLA-F mRNA while maintaining steady protein levels, indicative of virus-specific regulatory mechanisms. Moreover, we identified a soluble form of HLA-F secreted following Poly(I:C) stimulation. These findings reveal that HLA-F is dynamically regulated in trophoblasts during viral challenge and type I IFN signaling activation, supporting its broader immunomodulatory role in antiviral defense and immune tolerance at the MFI. Full article

Background/Objectives: Due to the utility of knowing the pathway involvement of metabolites detected in biological experiments, knowledgebases such as the Kyoto Encyclopedia of Genes and Genomes (KEGG), Reactome, and MetaCyc have annotated compound entries to specific pathways defined by the knowledgebase. However, these compound-pathway annotations are largely incomplete and are costly to obtain experimentally or curate from published scientific literature. This metabolite-pathway annotation incompleteness problem is amenable to machine learning (ML)-based solutions. But to date, no machine learning model has been trained on all three knowledgebases to maximize its performance and robustness. This may be due to inconsistencies in chemical structure representation that can confuse a model and greatly reduce generalizability. Methods: We constructed a new training dataset with roughly 50,000,000 entries using compound-pathway annotations derived from KEGG, Reactome, and MetaCyc. We trained and tested a multitask classification, graph convolutional neural network-like model that classifies compound involvement with 8056 pathways that have unique pathway representations, based on annotated compound chemical structures represented with chemical substructure features. While the initial dataset contained inconsistencies in chemical structure representations across knowledgebases, we alleviated this issue by standardizing chemical structure representation using InChI (IUPAC International Chemical Identifier) canonicalization. We compared the performance of the non-standardized versus the standardized dataset and quantified their generalizability by comparing training set compounds to their knowledgebase cross-references. Results: While the non-standardized dataset scored a mean Matthews correlation coefficient (MCC) of 0.8725 ± 0.0064, the standardized dataset scored an MCC of 0.9036 ± 0.0033. When comparing model generalizability, the non-standardized chemical structure representations had a huge 0.2687 drop in mean MCC, while the standardized chemical structure representations only had a 0.0384 drop in mean MCC. Conclusions: We constructed the largest ML-ready dataset for predicting compound-pathway involvement to date. Next, we constructed, trained, and evaluated the highest performing ML model capable of predicting the highest number of pathway annotations to date. We discovered that standardizing chemical structure representation is an essential step when predicting novel chemical structures. Full article

Background and Objectives: MicroRNAs (miRNAs) are small non-coding RNAs that regulate gene expression and have emerged as potential biomarkers in type 2 diabetes mellitus and its complications. This pilot exploratory study aimed to identify circulating miRNAs with differential expression in plasma from patients with newly diagnosed type 2 diabetes mellitus compared to age- and sex-matched healthy controls. Materials and Methods: Peripheral venous blood samples were collected from diabetic patients (n = 24) and controls (n = 12). Due to the exploratory nature of the study and limited sample material, samples were pooled within each group prior to plasma separation. Total RNA, including miRNAs, was extracted from plasma and analyzed using a high-throughput qPCR panel. Two normalization methods were applied to assess miRNA expression, and overlapping results were used for downstream analysis. Fold regulation was calculated using the 2^(−ΔCt) method. Results: A total of 33 and 42 miRNAs were identified as differentially expressed using the first and second normalization methods, respectively. Fourteen miRNAs were consistently downregulated across both methods. Several of these miRNAs, including hsa-miR-26a-5p, hsa-miR-146a-5p, hsa-miR-186-5p, hsa-miR-19a-3p, and hsa-miR-652-3p, have been previously associated with glucose metabolism, inflammation, and diabetic complications, such as retinopathy, neuropathy, and endothelial dysfunction. The pooling strategy enabled an efficient exploratory assessment of miRNA expression patterns while reducing inter-individual variability. Conclusions: This exploratory pilot study identifies a panel of circulating miRNAs with altered expression in pooled plasma samples from patients with newly diagnosed type 2 diabetes mellitus. These findings provide preliminary insights that warrant further validation in larger, individual-level studies to assess their diagnostic and prognostic potential. Full article

Myalgic Encephalomyelitis (ME) is a chronic, multisystem disease characterized by systemic metabolic dysfunction and post-exertional malaise (PEM). In this study, we investigated the dysregulation of irisin, an exercise-induced myokine, and its potential antagonism by thrombospondin-1 (TSP-1). In a cross-sectional study (92 ME patients vs. 44 sedentary healthy controls), plasma irisin and TSP-1 levels were measured at baseline and after a 90 min mechanical stress challenge applied to induce PEM. ME patients exhibited significantly lower baseline irisin (p < 0.05) and a blunted exertional response (p < 0.05). Paradoxically, baseline irisin was an independent predictor of fatigue severity (β = 0.728, p = 0.018), with moderate-to-severe patients showing elevated levels of both irisin and TSP-1 (p < 0.05), suggesting a compensatory but ineffective response. Functional cellular dielectric spectroscopy indicated that TSP-1 inhibits irisin signaling in a concentration-dependent manner. Irisin signaling was markedly reduced by both αvβ5 blockade and HSP90α inhibition in this experimental system, consistent with a diminished ability to counteract TSP-1. Collectively, these findings support a model in which dysregulation of the irisin–TSP-1 axis contributes to metabolic dysfunction in ME. Elevated circulating TSP-1 levels are associated with symptom severity and are linked to impaired irisin signaling in an HSP90α- and αvβ5-dependent context. This interaction is consistent with defective metabolic adaptation and highlights a potential therapeutic target that warrants further validation to restore energy homeostasis. Full article

Coagulase-negative staphylococci (CoNS) and mammaliicocci (MA) are common in food-derived samples and may act as antimicrobial resistance (AMR) reservoirs. A previous study reported a high S. aureus prevalence in poultry meat. The objective of this study was to characterize the species diversity, antimicrobial resistance, and biofilm-forming capacity of CoNS/MA from the same food samples. Species identification, antimicrobial susceptibility testing, resistance gene detection, molecular typing, and biofilm formation assays were performed. One hundred and forty-eight non-repetitive CoNS/MA isolates were detected in 85% of samples, and 14 species were identified. The most prevalent species were S. epidermidis (18.2%), S. simulans (12.8%), S. saprophyticus (12.2%), S. warneri (11.5%), and M. lentus (10.1%). Most samples harbored one or two different species, although some showed higher diversity. Although 27.0% of isolates were pan-susceptible, 22.3% were multidrug-resistant (MDR), significantly associated with M. lentus and S. epidermidis. Methicillin resistance was found in 10 isolates, mainly in S. epidermidis (lineages ST9, ST59, ST88 and ST640). Biofilm formation was observed in 24.3% of isolates (some of them MDR) and was significantly associated with S. pasteuri and S. xylosus and with samples from supermarkets. No methicillin-resistant isolates were biofilm producers. These findings highlight the diversity of CoNS/MA in poultry meat and their role as AMR reservoirs and persistence factors, emphasizing their relevance in food safety. Full article