Search Results (1,145)

Background: Obesity is increasingly recognized as a complex condition characterized by the convergence of metabolic dysregulation and psychological vulnerability. Insulin resistance (IR) has been identified as a biological bridge linking metabolic imbalance with affective symptoms such as anxiety, depression, and disordered eating behaviors. Methods: Fifteen obese adults (mean age = 25 ± 4.3 years) were evaluated through clinical examination, anthropometric assessment (BMI), biochemical assays (fasting insulin, AST, ALT), and standardized psychological assessments (STAI, BDI-II). In parallel, a rapid systematic review (2019–2025) synthesized evidence on the association between IR, affective dysregulation, binge eating disorder (BED), and the clinical role of insulin-sensitizing or incretin-based therapies. Results: Participants exhibited marked hyperinsulinemia (M = 79 μU/mL, SD = 6.61) and elevated anxiety (STAI-Trait = 54.22 ± 22.4) and depression scores (BDI-II = 21.6 ± 7.5). Liver enzymes were within normal limits. Literature synthesis confirmed consistent associations between IR, mood symptoms, and BED, associated with biological processes including inflammation, HPA axis hyperactivity, and dopaminergic imbalance. Integrated treatment approaches combining cognitive-behavioral therapy, medical nutrition therapy, and insulin-sensitizing agents (metformin, GLP-1RA, and GLP-1/GIP RA) were supported as effective and safe options. Conclusions: The coexistence of insulin resistance and emotional dysregulation in obesity is consistent with the hypothesis of a bidirectional metabolic–emotional axis. Early, integrated interventions addressing both metabolic and psychological domains may improve clinical outcomes and reduce progression toward chronic metabolic and psychiatric comorbidity. Full article

Background/Objectives: Rapid and accurate pathogen identification and antimicrobial susceptibility testing (AST) are critical for the proper management of patients with bloodstream infection (BSI). Real-time whole-genome sequencing (WGS) represents an attractive opportunity for exhaustive pathogen identification and antimicrobial susceptibility prediction (ASP). This feasibility study introduces BacT-Seq, a WGS-based prototype assay for the rapid and accurate identification of pathogens and the prediction of antimicrobial susceptibility from positive blood cultures using Oxford Nanopore Technologies (ONT) sequencing. Methods: A total of 200 positive blood culture samples from patients with a confirmed BSI were included in this study. DNA isolation from blood cultures was optimized prior to GridION (ONT) sequencing. Pathogen identification and several ASP methods were compared to conventional identification and phenotypic AST methods. Results: Most of the mono-microbial (89%) and poly-microbial (88%) samples were identified by BacT-Seq in less than 10 min of sequencing. While identification of poly-microbial samples remains challenging, identification of mono-microbial samples by sequencing was non-inferior to that of the conventional approach, even revealing an added value in terms of exhaustivity and/or taxonomic resolution. Machine-learning-based ASP models yielded 80% predictions in 2.5 h of sequencing. Their ability to predict resistance phenotypes varied with the microbial species evaluated, from 55/57 (96.5%) for Escherichia coli to 24/48 (50.0%) for Pseudomonas aeruginosa. Conclusions: This study demonstrates the feasibility of implementation of the BacT-Seq platform for the fast and accurate identification of pathogens from positive blood cultures. BacT-Seq performance of resistance predictions by bioinformatics tools is promising but requires further optimization and validation before clinical implementation. Full article

Background/Objectives The leopard (Panthera pardus), an apex predator listed in CITES Appendix I and classified as Vulnerable by the IUCN, is undergoing severe population declines driven by habitat loss, human–wildlife conflict, and illegal trade. Rapid and reliable species and individual identification is critical for conservation and forensic applications, particularly when analyzing highly processed or degraded seized wildlife products, where morphological identification is often impossible. We aimed to develop and validate a robust multiplex quantitative real-time PCR (qPCR) assay combined with a short tandem repeat (STR) system for the species-specific detection and individual identification of P. pardus. Methods The qPCR assay (Ppar Qplex) was designed to target a mitochondrial Cytochrome b (Cyt b) fragment for species confirmation, a nuclear marker (PLP) for general Feliformia detection and quantification, and an artificial internal positive control (IPC) to monitor PCR inhibition. The assay’s performance was validated for robustness, specificity, sensitivity, repeatability, and reproducibility, utilizing DNA extracted from 30 P. pardus individuals (hair and feces) and tested against 18 related Feliformia species and two outgroups. Individual identification was achieved using a set of 18 STR loci and a sex determination system adapted from previously published Panthera panels. Results Validation demonstrated high specificity for the Ppar Qplex: mitochondrial amplification occurred exclusively in P. pardus samples. The nuclear marker consistently amplified across all 18 tested Feliformia species but not the outgroups. The assay showed high analytical sensitivity, successfully detecting DNA at concentrations as low as 1 pg/µL, with consistent results confirmed across different sample types, replicates, and independent users. Furthermore, the STR multiplex successfully generated 30 unique individual profiles using the 18 polymorphic loci and the sex determination system. Conclusions The combined qPCR assay and STR system provide a fast, sensitive, and highly specific molecular framework for rapid leopard detection, quantification, and individual identification from a wide range of sample types. These tools strengthen forensic capacity to combat wildlife crime and provide critical data to support evidence-based conservation management of P. pardus. P. pardus, an apex predator listed in CITES Appendix I and classified as Vulnerable by the IUCN, is undergoing severe population declines driven by habitat loss, human–wildlife conflict, and illegal trade. Rapid and reliable identification of seized specimens is therefore critical for conservation and forensic applications, mainly when products are highly processed. We developed and validated a multiplex quantitative real-time PCR (qPCR) assay targeting the mitochondrial gene Cytochrome b (Cyt b) for species-specific detection. The assay was tested on verified leopard individuals and validated across 18 Feliformia and two outgroup species (Homo sapiens, Canis lupus familiaris). Analytical performance was assessed through robustness, specificity, sensitivity, repeatability, and reproducibility. Mitochondrial amplification occurred exclusively in leopard samples, while nuclear markers amplified consistently across Feliformia but not in outgroup species. The assay’s limit of DNA detection is 1 pg/µL and produces consistent results across replicates, tested types of samples (hair, feces), and independent users, with internal controls confirming the absence of inhibition. In addition, we present the results of successful individual identification using the set of 18 STR loci and the sex determination system. The developed qPCR and STR systems provide a fast, sensitive, and specific solution for leopard detection and quantification, reinforcing forensic efforts against wildlife crime and supporting conservation of P. pardus. Full article

The growing demand for robust food authentication methods has driven the establishment of fast, sensitive, and field-based detection systems for identifying meat species. This study presents a colorimetric-based PSR approach for identifying yak (Bos grunniens) meat within fresh, thermally processed, and blended meat samples. Targeting the mitochondrial D-loop locus, the assay incorporates a simple alkaline lysis (AL) procedure for efficient DNA extraction, eliminating the requirement for specialized instrumentation. The PSR assay demonstrated high specificity, showing no evidence of cross-reactivity with closely associated food animals such as buffalo, cattle, goat, sheep, mithun, and pig. Sensitivity assessment revealed the assay’s capability to detect 1 pg of yak DNA, with reliable performance in samples exposed to thermal conditions up to 121 °C. Additionally, the technique detected yak meat down to a concentration of 0.1% in binary beef mixtures. This method provides a significant improvement in sensitivity over end-point PCR and is particularly well-suited for field applications due to its practical simplicity, affordability, as well as no reliance on sophisticated instrument. This is, to the best of our understanding, the first reported PSR-based approach developed for the identification of yak meat, offering a robust tool for food origin verification, regulatory enforcement, and product integrity monitoring. Full article

Duckweed (Spirodela polyrhiza), a fast-growing aquatic plant rich in protein and bioactive compounds, offers a sustainable alternative to conventional aquafeed protein sources. This study evaluated the effects of incorporating 25–75% duckweed meal into a commercial feed on grass carp (Ctenopharyngodon idella) over a 6-week trial. Fish meal, wheat starch, and vegetable oil was added in amounts to obtain isonitrogenous and isoenergetic diets. Additionally, another grass carps were used for extended feeding until they reached approximately 1000 g, using the feed with the optimal duckweed inclusion rate (25%). Fish fed a diet consisting of 75% commercial feed and 25% duckweed meal (F75D25) exhibited significantly higher weight gain. Muscle analysis revealed increased protein content (up 15%, p < 0.05) and improved amino acid and fatty acid profiles. Liver, muscle, and blood assays showed elevated antioxidant enzyme activities (SOD up 20%, LYS up 18%; p < 0.05) and immune markers (CRP, GOT; p < 0.05), indicating enhanced health status. Transcriptomic and metagenomic analyses confirmed the upregulation of immune-related genes (e.g., SOD1, IL-6; fold change > 2, p < 0.01) and beneficial shifts in gut microbiota (e.g., increased Firmicutes). These findings highlight duckweed’s potential as a nutrient-rich, health-promoting ingredient for sustainable aquaculture diets. Full article

Precision medicine starts with a precision diagnosis. Yet up to 80% of cases of monogenic diabetes, a form of diabetes characterized by mutations in a single gene, are either overlooked or misdiagnosed. A genetic test for monogenic diabetes does not always lead to a precise diagnosis, as novel variants are often classified as variants of unknown significance. Variant interpretation requires collation of a framework of evidence, including population, computational, and segregation data, and can be assisted by functional analysis. The inclusion of functional data can be challenging, depending on the number of benign and pathogenic variants available for benchmarking assays. Glucokinase is the rate-limiting step for glucose metabolism in the pancreatic beta-cell and governs the threshold for glucose-stimulated insulin release. Loss-of-function alleles in the glucokinase (GCK) gene are a cause of stable fasting hyperglycemia from birth and/or diabetes. In this study, we functionally characterized 25 variants identified during diagnostic testing or in exome sequencing studies. We assessed their kinetic characteristics, stability, and interaction with pharmacological and physiological regulators. We integrated our functional data with existing data from the ClinGen Monogenic Diabetes Variant Curation Expert Review panel using a gene-specific framework to assist variant classification. We show how functional evidence can aid variant classification, thus enabling diagnostic certainty. Full article

Photosynthetic prokaryotes known as cyanobacteria produce an extensive range of primary and secondary metabolites that serve multiple biotechnological and biomedical purposes. The non-model filamentous Phormidium species capture researchers’ attention through their biotechnological potentials from diverse metabolites and their ability to thrive in tough environments while producing bioactive compounds. In this study, a thermotolerant strain of Phormidium ambiguum was isolated from the Chundzha thermal springs in southeastern Kazakhstan and characterized morphologically, physiologically, and biochemically. This cyanobacterium demonstrated fast growth in its culture media along with significant accumulation of proteins (44.8% DM), carbohydrates (45% DM), and photosynthetic pigments like chlorophyll a and valuable carotenoids, including b-carotene, myxoxantophyll and zeaxanthin. The LC-ESI-MS/MS analysis of cyanobacterial non-polar extract identified 150 potential metabolites which include fatty acid derivatives, terpenoids and carotenoid-related compounds known for their antioxidant and antimicrobial properties, as well as immune system stimulation. Biological assays confirmed a weak antioxidant capacity in the DPPH test, while in immunological assays, the extract of P. ambiguum stimulated T lymphocyte proliferation and activation, as well as NK cell proliferation in vitro. It also exhibited moderate antibacterial activity towards tested Gram-negative and Gram-positive bacterial strains. While additional studies are required to address environmental robustness, biosynthetic regulation, and practical scalability, the present findings indicate that P. ambiguum represents a valuable non-model cyanobacterium for exploratory bioprospecting. Its metabolite profile and observed bioactivities support further investigation of this thermotolerant strain as a potential source of immunomodulatory, antioxidant, and antimicrobial compounds under controlled conditions. Full article

Background/Objectives: Wound healing requires simultaneous pain control, inflammation management, infection prevention, and tissue regeneration. This study aimed to develop and evaluate in vitro a non-contact thermosensitive spray hydrogel combining lidocaine for rapid analgesia and allantoin for tissue repair. Methods: The effects of chitosan and Poloxamer 407 on viscosity, spray diameter, and bioadhesion ability of hydrogels were optimized using response surface methodology. Lead formulations (S1 and S2) were selected via a desirability function within the software. The pH, gelation temperature (T_G), rheological behavior, sprayability, bioadhesion, and lidocaine release using the dialysis bag method were assessed. The in vitro cytotoxicity, anti-inflammatory activity (TNF-α), and cell migration (scratch assay) of the formulations were investigated. Results: The viscosity values (42.7–58.7 mPa·s) indicated suitability for spraying at room temperature. T_G was 28.7 ± 0.6 °C (S1) and 29.3 ± 0.3 °C (S2), enabling rapid sol–gel transition at skin temperature. The lidocaine release reached 95–100% within 120 min. S2 exhibited lower viscosity and wider spray diameter, improving applicability on larger wound areas. In vitro cytotoxicity, scratch assay, and inflammatory marker analyses demonstrated that the optimized sprayable hydrogels exhibited a biocompatible and cell-healing profile. Conclusions: The developed thermosensitive spray hydrogel enables the combined delivery of lidocaine and allantoin, rapid gelation at body temperature, and touch-free administration. Its suitable viscosity and sprayability, and fast lidocaine release profile indicate high patient compliance and a significant advantage over conventional cream/ointment formulations, particularly regarding painless application, reduced contamination risk, enhanced therapeutic potential, and confirmed in vitro biocompatibility with supportive effects on keratinocyte behavior. Full article

Neuraminidase (NA) decorates the surface of the influenza virus, exerting a sialidase activity that enables the viral particle to be released in the host cell. Numerous sialic-based antiviral agents competitively bind to the NA cavity and are marketed worldwide for the treatment of Influenza A infection. We designed and validated a structure-based pharmacophore model for influenza neuraminidase (NA), which guided a virtual screening campaign against an in-house library of compounds already available for testing. This fast and cost-effective in silico strategy resulted in the identification of seven candidates possessing indole or isoquinoline chemical core. In vitro assays confirmed their favorable cytotoxicity profiles and identified only one, the 1-(1H-indol-3-ylcarbonyl)-3-piperidinecarboxylic acid (1), with reproducible inhibitory activity toward NA at non-cytotoxic concentrations. This work suggested a validated workflow for the discovery of novel NA inhibitors and highlighted an indole-based hit compound as a starting point for further optimization. Full article

Water convolvulus (Ipomoea aquatica Forssk.) is a fast-growing leafy vegetable valued for its nutritional and antioxidant properties; however, suboptimal seed physiology can hinder its germination and early growth. Non-thermal plasma (NTP) treatment is an eco-friendly seed-priming method that enhances seed performance and crop quality without the use of chemical inputs. This study evaluated the effects of NTP exposure (0, 5, 10, and 20 min) using a dielectric barrier discharge (DBD) plasma with an air gas flow rate of 1.5 lpm on the germination, seedling growth, pigment and protein content, nitrogen assimilation, and antioxidant capacity of water convolvulus. Plasma treatment of seeds increased germination in a time-dependent manner. The surface hydrophilicity improved with increasing treatment time. Seedlings grown from seeds treated for 10 min exhibited longer shoots (+10.1%) and roots (+17.8%). The shoot nitrate content increased by 66.3%. At 10 min, the total phenolics and flavonoids increased by 26.5% and 37.2%, respectively, with an accompanying increase in antioxidant activity, as measured by DPPH, ABTS, and FRAP assays. These findings demonstrate that a 10 min NTP treatment of seeds improves germination, growth, nutrient assimilation, phytochemical accumulation, and antioxidant activity in water convolvulus seedlings, highlighting its potential as a sustainable and chemical-free seed-priming technology with considerable potential to enhance the productivity and nutritional quality of plant microgreens in modern agriculture. Full article

An indirect, novel, fast, and facile assay was developed for the colorimetric determination of fluoride anions using 96-well plates. The proposed method relies on the colorimetric degradation caused by fluoride ions after their reaction with the iron–thiocyanate complex in an acidic medium. The procedure required the addition of minimal amounts of ferric iron and thiocyanate anion solutions to form the corresponding complex with an intense blood-red color, after which, upon addition of fluoride ions, this complex would dissociate, and its color would gradually fade depending on the analyte concentration. The colorimetric differences were measured using a simple imaging device such as a flatbed scanner. Various parameters affecting the analytical performance of the proposed method were optimized, including solution concentrations, pH values, and reaction time for Fe(III)-SCN complex formation and its disintegration process. The proposed assay was successfully applied to the determination of F⁻ in oral hygiene product samples. The method exhibited acceptable detection limits (3.2 mg L⁻¹) with sufficient precision, good intra-day and inter-day reproducibility (ranging from 1.5 to 5.2%), and high selectivity against other anions and components of the samples under study. Full article

Avocado (Persea americana Mill.) is cultivated in a wide range of environments, from tropical and semitropical to subtropical regions. Its fruit, of high nutritional value, is increasingly demanded worldwide. Spain is the main European producer, but avocado cultivation in certain areas, such as the Canary Islands, requires the genetic identification of West-Indian rootstocks because they often show tolerance to low-quality water and soil salinization. In the present study, eight novel Retrotransposon-Based Insertion Polymorphism assays, derived from previously characterized inter-Primer Binding Site markers, have been developed and evaluated by multiplex PCR across 58 P. americana cultivars. The results showed 100% specificity and sensitivity in detecting the West-Indian genomic component, both in pure and hybrid avocado cultivars. This cost-effective and fast molecular tool provides a valuable resource for characterization and selection programs of avocado cultivars genetically related to the West-Indian horticultural race. Full article

Microneedle array-based drug delivery offers a minimally invasive and safe approach for breaching the skin barrier, enabling localized and targeted treatment—an advantage particularly valuable in chronic condition management, such as rheumatoid arthritis (RA). RA presents a multifaceted pathophysiology, often necessitating long-term pharmacological management. However, conventional oral administration may lead to systemic drug distribution, increasing the likelihood of adverse effects, and ultimately undermining therapeutic efficacy. In this study, a hollow microneedle array was employed for effective delivery of Tofacitinib and the antioxidant N-acetylcysteine (NAC). A comprehensive evaluation was conducted across multiple levels, in which inflammation and cartilage degradation were assessed histologically using hematoxylin-eosin (H&E) and Safranin O–Fast Green staining. Radiologically, micro-computed tomography (micro-CT) was employed to visualize bone structure alterations. On the molecular level, enzyme-linked immunosorbent assay (ELISA) was used to quantify inflammatory cytokines and oxidative stress markers. Furthermore, differentially expressed genes and enriched signaling pathways were identified through transcriptomic profiling pre- and post-treatment. And the potential regulatory targets and mechanistic insights into the therapeutic response were elucidated through correlation analyses between gene expression profiles and pathological indicators. This study provides a mechanistic and computational basis for precision targeted therapy, validates the efficacy and safety of microneedle delivery in a rheumatoid arthritis (RA) model, and demonstrates its potential application in local drug delivery strategies. Full article

Cardiac troponin I (cTnI) is a critical biomarker for the diagnosis of acute myocardial infarction (AMI), but conventional detection methods are often time-consuming and require specialized laboratory equipment. To meet the need for rapid and feasible detection, there is an urgent demand for methods that are fast, specific, and easy to use. In this study, two aptamers (Tro4 and Tro6), which specifically bind to different epitopes of cTnI, were employed to construct a dual-aptamer sandwich system on a lateral flow assay (LFA) strip. The test strip can deliver results within 10 min and shows a detection limit of 11.70 ng·mL⁻¹. It also exhibited excellent stability after storage at room temperature for up to four months. The assay demonstrated high analytical accuracy, as evidenced by recovery rates from spiked serum samples ranging from 95.11% to 103.17%. These results suggest that the proposed aptamer-based LFA is highly suitable for rapid screening of cTnI, especially in point-of-care settings and resource-limited environments. From a diagnostic perspective, this method holds great promise for improving the timely detection and management of AMI and other myocardial injuries. Full article

Introduction/Objectives: Ginsenoside F1, a pharmacologically active saponin derived from Panax ginseng, exhibits diverse bioactivities, but its use is limited because it is difficult to purify and has high production costs. To overcome these challenges, a ginsenoside F1-enriched extract named SGB121 was developed. This study aimed to evaluate the therapeutic efficacy of SGB121 in a high-fat, high-carbohydrate (HFHC) diet-induced metabolic dysfunction-associated fatty liver disease (MAFLD) mouse model and to elucidate its mechanism of action using F1-based cellular assays. Methods: Male C57BL/6 mice (6 weeks old) were fed an HFHC diet to induce MAFLD and were treated with SGB121. Hepatic lipid accumulation, oxidative stress markers, and metabolic parameters were analyzed. In parallel, human hepatocellular carcinoma (HepG2) cells exposed to free fatty acids (FFAs) were used to assess oxidative stress and lipid accumulation. Mechanistic studies were conducted using purified F1 to examine adenosine monophosphate-activated protein kinase (AMPK) activation and related pathways. Results: SGB121 reduced hepatic lipid accumulation, malondialdehyde (MDA) levels, and fasting insulin while restoring glutathione (GSH) content and improving the homeostasis model assessment of insulin resistance (HOMA-IR) in MAFLD mice. In FFA-treated HepG2 cells, both SGB121 and F1 decreased reactive oxygen species (ROS), suppressed sterol regulatory element-binding protein 1 (SREBP1), enhanced peroxisome proliferator-activated receptor-α (PPARα) and β-oxidation, and restored insulin receptor substrate (IRS)/protein kinase B (Akt)/glucose transporter 2 (GLUT2) signaling. Conclusions: SGB121 ameliorates MAFLD and related metabolic dysfunction through antioxidant, lipid-regulating, and insulin-sensitizing actions, highlighting its potential as a safe multifunctional nutraceutical for MAFLD management. Full article