Search Results (538)

CRISPR–Cas9 has progressed from an experimental tool to a therapeutic modality, marked by the first regulatory approvals of an ex vivo-edited autologous CD34+ hematopoietic stem cell product that induces fetal hemoglobin (CASGEVY/exa-cel). In this narrative review, we synthesize modality-specific molecular diagnostic strategies used across early CRISPR clinical translation. In parallel, early clinical experience has begun to demonstrate the feasibility of in vivo editing, including subretinal delivery for CEP290-associated inherited retinal degeneration (EDIT-101 programme) and hepatocyte-targeted lipid nanoparticles (LNPs) for liver-derived targets such as transthyretin and plasma prekallikrein (KLKB1). As translation expands across hematologic, metabolic, ocular and oncology indications, development is increasingly constrained by the predictability and safety of editing outcomes, delivery-determined biodistribution and exposure time, and immune recognition of bacterial Cas9 orthologs and delivery components. We summarize diagnostic readouts for confirming patient genotype, quantifying on-target editing and expression changes, assessing off-target and structural outcomes using orthogonal assays, and monitoring clonal dynamics and immune responses during long-term follow-up. We also discuss how these readouts interface with CMC controls and regulatory expectations for advanced therapy medicinal products (ATMPs), highlighting the need for fit-for-purpose, standardized testing frameworks in early trials. Full article

Non-motor symptoms of PD impair quality of life and remain challenging to treat. Here, we examined the effects of short- (38 days) and long-term (178 days) supplementation with the natural polyamine spermidine on anhedonia and anxiety-like behaviours in a 6-hydroxydopamine-induced rat model of PD and linked them with spermidine’s anti-inflammatory properties. Behavioural assessments (cylinder, sucrose preference, elevated plus-maze tests) were conducted during progressive neurodegeneration and after oral treatment. Under the same conditions, peripheral inflammation was evaluated by the total leukocytes and their subpopulation numbers (hematological analysis) and by CD4⁺ and CD8⁺ T lymphocyte percentages (imaging flow cytometry); the plasma levels of interleukins 4 and 10 and corticosterone (enzyme-linked immunosorbent assay) were also evaluated. The safety of long-term supplementation was assessed using standard biochemical markers (chemistry analyser). Both treatment regimens reversed 6-hydroxydopamine-induced lymphopenia. Long-term spermidine treatment increased the number of TCD4⁺ lymphocytes and monocytes and elevated the plasma concentrations of IL-4 and IL-10, while reducing corticosterone levels. These immunomodulatory effects were associated with reduced anhedonia and anxiety. All of the biochemical safety parameters remained within normal ranges. Spermidine alleviates neuropsychiatric symptoms in a rat model of progressive neurodegeneration in the nigrostriatal system through its regulatory influence on peripheral immune responses. Exploring the systemic mechanisms underlying spermidine’s effects could unveil innovative supplementation strategies and expand treatment options for managing symptoms in PD. Full article

Inflammatory myocardial involvement has been reported in canine leishmaniosis (CanL); however, studies evaluating the degree of myocardial dysfunction in affected dogs are limited. This prospective study aimed to investigate myocardial involvement in dogs with CanL using conventional and speckle-tracking echocardiography (STE), focusing on the assessment of left ventricular systolic function and myocardial strain. Symptomatic, initially untreated dogs with a diagnosis of leishmaniosis and free from other vector-borne diseases or underlying heart diseases were enrolled (Leish group). Healthy dogs matched for age, body weight, breed, and sex were selected for the control group (C group). At the time of inclusion (T0) and at each follow-up, laboratory tests as well as conventional echocardiographic examination and STE were performed. For strain analysis, apical longitudinal long-axis 4-chamber, 3-chamber, and 2-chamber views were used (2C, 3C, 4C, respectively) to obtain the average global longitudinal strain (GLSAV), which is recognised to have the maximum reliability as an indicator of left ventricular dysfunction in humans. The software obtains GLSAV by averaging the longitudinal strain values from all left-ventricular segments derived from the multiple apical views. After enrolment, dogs were treated with a combination of meglumine and allopurinol and were monitored for six months. Clinical-pathological and echocardiographic data were collected at follow-up at 1, 3, and 6 months after the start of treatment (T1, T2, T3) and compared between the two study groups using appropriate statistical tests. Sixteen dogs composed the C group and nine dogs the Leish group. At T0, none of these dogs had abnormalities in cardiac auscultation, plasma cardiac troponin concentration was within the reference range, and standard echocardiographic examination excluded underlying cardiac diseases. The comparison between C and Leish groups did not show a statistically significant difference in any of the strain parameters analysed (GLSAV, GLS4C, GLS3C, GLS2C). Moreover, strain values in the Leish group did not change significantly over time. In conclusion, in this preliminary study on a limited population of dogs with leishmaniosis, both conventional echocardiography and STE failed to reveal clear changes suggestive of left ventricular systolic dysfunction secondary to possible myocarditis or as a consequence of the systemic disease in dogs with active leishmaniosis. However, further STE studies in larger cohorts of dogs with leishmaniosis are needed to confirm and expand our findings. Full article

Obesity is a chronic, highly prevalent disease affecting nearly one-third of the global population and represents a major independent risk factor for heart failure (HF), particularly heart failure with preserved ejection fraction (HFpEF). Excess adiposity—especially visceral and epicardial adipose tissue (EAT)—acts as an active endocrine and immune organ, promoting chronic low-grade inflammation, oxidative stress, endothelial dysfunction, and adverse myocardial remodeling. Expanded EAT exerts both paracrine inflammatory effects and mechanical constraint on the myocardium, contributing to diastolic dysfunction, microvascular impairment, atrial arrhythmogenesis, and elevated filling pressures despite preserved systolic function. Evidence demonstrates a dose–response relationship between increasing body mass index and incident HF. Clinically, obesity-related HFpEF is characterized by concentric left ventricular hypertrophy, impaired relaxation, increased plasma volume, reduced exercise tolerance, and relatively low natriuretic peptide levels, complicating diagnosis. HF management includes traditional treatment with diuretics, renin-angiotensin system inhibitors, β-blockers, mineralocorticoid receptor antagonists, and angiotensin receptor-neprilysin inhibitors. These agents widely remain foundational as they primarily target hemodynamic and neurohormonal pathways in HF. In contrast, sodium–glucose cotransporter 2 inhibitors consistently reduce HF hospitalizations across the ejection fraction spectrum, while glucagon-like peptide-1 receptor agonists and dual incretin therapies (e.g., tirzepatide) promote substantial weight loss, improve symptoms, and demonstrate promising anti-remodeling effects in obesity-related HFpEF. Recognizing obesity-driven HF as a distinct cardiometabolic entity supports an integrated therapeutic strategy combining structured weight reduction with guideline-directed HF polypharmacotherapy to address both hemodynamic burden and upstream adiposity-related mechanisms. Full article

The management of multiple sclerosis (MS) is shifting from a phenotype-based framework toward a biologically driven precision medicine model, as conventional magnetic resonance imaging (MRI) inadequately captures smoldering inflammation and progression independent of relapse activity (PIRA). This systematic review aimed to synthesize current evidence on the diagnostic and prognostic utility of fluid biomarkers in distinguishing acute inflammatory injury from chronic neurodegeneration. A comprehensive search of Web of Science, PubMed, and Scopus (January 2020–September 2025) identified 28 eligible studies including 7775 participants (6365 MS patients and 1410 controls). Biomarkers derived from serum, plasma, cerebrospinal fluid (CSF), and stool were evaluated in relation to clinical disability measured using the Expanded Disability Status Scale (EDSS) and magnetic resonance imaging (MRI) outcomes. Neurofilament light chain (NfL) consistently predicted acute inflammatory activity, gadolinium-enhancing lesions, and relapse-associated worsening, but levels were reduced by high-efficacy therapies and did not reliably predict PIRA. In contrast, glial fibrillary acidic protein (GFAP) was associated with astrogliosis, disability progression, and retinal thinning, even in patients with low inflammatory activity. Additional CSF, metabolic, and immunologic markers correlated with neurodegeneration and disease severity. Nevertheless, broader clinical use will require greater assay standardization, improved consistency across cohorts, and validation in prospective longitudinal studies. These findings compel a shift toward a multi-biomarker model to guide personalized therapeutic strategies and develop targeted neuroprotective treatments for progressive multiple sclerosis. Full article

Background/Objectives: Clopidogrel is a widely prescribed antiplatelet prodrug that requires bioactivation, primarily by the polymorphic CYP2C19 enzyme. Genetic variation in this enzyme leads to differences in active metabolite formation and has prompted the development of pharmacogenetics-guided prescribing. However, current pharmacogenetic strategies are grounded in drug metabolism knowledge derived from mass balance studies conducted in small groups of healthy volunteers. This narrow evidence base may limit the data’s applicability to real-world settings, where factors like polypharmacy or altered organ function may influence drug response. Methods: Pharmacogenetics could benefit from real-world drug metabolism and excretion studies, which we conducted for clopidogrel in 38 kidney and 16 liver transplant recipients from the TransplantLines Biobank and Cohort Study (NCT03272841), utilizing existing LC-SWATH/MS pharmacometabolomic data. Clopidogrel-associated metabolic signals were identified using xenobiotic metabolism knowledge and literature-reported pathways. Results: Across both transplant groups, 26 clopidogrel-associated features were prioritized, of which some matched previously reported urinary metabolites, had previously been observed in plasma, or represented previously unreported metabolites. Clopidogrel carboxylic acid predominated in kidney transplant recipients, whereas its glucuronide form was most abundant in liver transplant recipients. Notably, unmetabolized clopidogrel was consistently detected across all patients. Moreover, our data support a thiol desulfurization route, aligning with emerging evidence of clopidogrel’s role as a hydrogen sulfide-releasing drug. Conclusions: More (putative) clopidogrel metabolites were detected than previously reported, demonstrating the value of pharmacometabolomics in expanding our understanding of drug metabolism. This approach provides novel data that may complement pharmacogenetics research to understand clopidogrel response variability among treated patients. Full article

Background: Valproic acid (VPA) poisoning has a dynamic clinical course and may require extracorporeal toxin removal (ECTR) in severe cases. Intermittent hemodialysis is the preferred ECTR technique; however, clinical experience with expanded hemodialysis (HDx) using medium cut-off (MCO) membranes in acute VPA intoxication is scarce. We describe a case of severe VPA poisoning managed with intermittent HDx and outline the clinical rationale and kinetic response. Case Report: A 54-year-old woman presented to the emergency department after accidental presumably ingesting approximately 4 g of VPA, with depressed consciousness (Glasgow Coma Scale 7) and metabolic acidosis (pH 7.10, HCO₃⁻ 13 mmol/L, PCO₂ 50 mmHg, lactate 2.8 mmol/L, ionized calcium 0.8 mmol/L, elevated anion gap). Initial plasma VPA was 262.99 µg/mL, ammonia was 14 µmol/L, and cranial computed tomography showed no acute abnormalities. ECTR was initiated in the intensive care unit as intermittent HDx using an MCO dialyzer for 4 h. Serial VPA concentrations were obtained before treatment, at 2 h, and at the end of the session to guide real-time prescription adjustment, with an increase in blood flow from 200 to 230 mL/min. Results: VPA decreased from 262.99 µg/mL pre-HD to 141.48 µg/mL at 2 h (46.2% reduction) and 97.81 µg/mL at 4 h (62.8% reduction), with clear improvement in the level of consciousness. A mild post-dialysis rebound was observed (100.07 µg/mL at 14 h). The patient recovered without additional ECTR and was discharged with normalized VPA levels on follow-up. Conclusions: In this patient, intermittent HDx with an MCO membrane was feasible, well tolerated, and associated with rapid VPA clearance and neurological recovery. Serial drug monitoring enabled bedside optimization of the dialysis prescription and post-treatment evaluation. A single HDx session was sufficient, and VPA therapy was safely reintroduced under close monitoring. Full article

ABC transporters (ATP-binding cassette transporters) constitute one of the largest known protein families and are widely distributed in plants. Their primary function involves utilizing energy derived from ATP hydrolysis to transport substrates across membranes against concentration gradients. These transporters play crucial roles in the translocation and accumulation of metabolites, stress tolerance, disease resistance, and plant defense. Lotus is an important traditional Chinese medicinal herb and contains active ingredients primarily composed of secondary metabolites, whose transport and accumulation require the involvement of ABC transporters. However, the function of these ABC transporters remains unexplored in lotus. In this study, 122 ABC transporter genes were predicted within the lotus genome. We identified 1~15 conserved motifs among the NnABC proteins and most of them were stable proteins predominantly located on the plasma membrane with ExPASy-ProtParam, ProComp and WoLF PSORT analysis. Phylogenetic tree analysis revealed that the lotus ABC transporter gene family could be divided into eight subfamilies, from ABCA to ABCI, and the evolution was predominantly driven by purifying selection. Comparative transcriptome analysis between the cultivar ‘Yindu Zhimi’ with orange-reddish stamen and ‘Weishan Hong’ with yellowish stamen, along with quantitative real-time PCR results, showed that the NnABCG25 gene is highly specifically expressed in the orange-reddish stamen. Molecular docking demonstrated that NnABCG25 has a stable affinity for lycopene, β-carotene and β-apocarotenal, suggesting its potential involvement in the transport of carotenoids in the stamen. These findings expand our understanding of the role of ABC transporters in the transport and accumulation of carotenoids, as well as providing a valuable reference for research on the ABC transporter gene family in other plants. Full article

Non-thermal plasma-driven advanced oxidation is a promising method for treating organic wastewater, which exhibits rapid reaction kinetics and high pollutant removal and does not need chemical reagents. However, its practical application is often limited by high specific energy consumption and the inefficient mass transfer of short-lived reactive species across the gas–liquid interface. This review summarizes the fundamentals of non-thermal plasma chemistry and the process intensification of plasma multiphase reactors by mass transfer enhancement and waste energy-driven conversion. This review focus on four coupling approaches: microbubble-assisted plasma to expand the reactive interfacial area; plasma coupled with hydraulic cavitation to enhance convection and radical formation; plasma–piezoelectric catalysis coupling to harvest hydraulic energy and promote charge-driven reactions; and plasma-assisted Fenton oxidation to improve the utilization of weakly oxidizing species (H₂O₂). The energy efficiency of various plasma-based oxidation systems is compared and discussed clearly. Key remaining challenges are also discussed, including standardized energy efficiency assessment, scale-up and hydrodynamic control, catalyst stability and fouling, by-product formation and toxicity, and long-term operational reliability. Overall, this review aims to provide guidance for developing efficient plasma-based wastewater treatment systems for large-scale applications. Full article

Alpha-2-macroglobulin (α₂M) is a conserved plasma glycoprotein traditionally known for its broad-spectrum protease inhibitory activity. However, emerging evidence indicates that its activated form, α₂M*, generated via proteolytic cleavage or nucleophilic attack, functions as a versatile signaling ligand. By engaging specific cell-surface receptors, most notably low-density lipoprotein receptor-related protein 1 (LRP1) and glucose-regulated protein 78 (GRP78), α₂M* orchestrates a diverse array of intracellular programs, including the PI3K/Akt/mTOR, MAPK/ERK, and JAK/STAT cascades, as well as mechanosensitive YAP/TAZ signaling. These pathways collectively govern fundamental cellular processes such as proliferation, metabolic reprogramming, cytoskeletal remodeling, and inflammatory adaptation across various cell types, including macrophages, cardiomyocytes, and malignant cells. Altogether, this review synthesizes current knowledge on α₂M activation, structural transitions, receptor interactions, and downstream signaling, highlighting the expanding functional landscape of α₂M* as a potent regulator of intracellular communication with implications for physiology and disease. Full article

Advances in regenerative medicine have positioned platelets and their derivatives—including platelet-rich plasma, platelet-rich fibrin, platelet lysate, extracellular vesicles, and purified growth factors—as promising interventions specifically for skin and bone aging, two clinically accessible tissues with robust preclinical and clinical evidence for platelet-derived component-based rejuvenation and regeneration. Because much of the available evidence comes from injury models or age-associated inflammatory/degenerative diseases, we explicitly distinguish pathology-targeted inflammation resolution/repair from rejuvenation under physiological aging. This review summarizes the composition and core bioactivities of platelet-derived products and delineates their putative anti-aging mechanisms, encompassing proangiogenic signaling, immunomodulation, attenuation of oxidative stress, regulation of extracellular matrix turnover, and stimulation of osteogenesis. We further evaluate emerging applications that expand therapeutic performance, such as platelet-mimetic delivery vehicles, engineered and sustained-release formulations, and targeted use of subcellular structures. Evidence from recent preclinical and clinical studies indicates favorable safety profiles and signals of efficacy across cutaneous rejuvenation and skeletal regeneration, while underscoring persistent challenges related to product standardization, dosing, and outcome measures. Collectively, platelet-based therapeutics represent a versatile platform with broad applicability to anti-aging interventions in skin and bone and strong potential for translation through continued bioengineering and clinical validation. However, because most available evidence comes from injury models or age-associated diseases (e.g., photoaging, chronic wounds, osteoarthritis, osteoporosis), direct extrapolation to physiological aging is limited; throughout, we explicitly contrast these contexts, specify their indication-specific endpoints, and summarize the main translational limitations. Full article

The main objective of this paper is to establish links between two different plasma kinetic theories. We start by summarizing a two-temperature kinetic theory of multicomponent magnetized reactive plasmas, where electrons and heavy species have their own temperature. The Knudsen number is taken to be proportional to the square root of the mass ratio, and polyatomic species are taken into account. We next summarize the one-temperature kinetic theory of multicomponent magnetized reactive plasmas when the mass ratio remains of order unity. The complex tensorial structure of the transport fluxes is addressed, as well as the symmetry properties of the multicomponent transport coefficients. We then establish new links between these two theories by using the two-temperature scaling in the transport linear system obtained from the one-temperature kinetic theory. By expanding the solutions of the transport linear systems in terms of the Knudsen number, the flux structure of the two-temperature theory is recovered from the one-temperature theory as well as the second-order corrector terms. We also address the solution of transport linear systems by using fast and convergent iterative algorithms and their improvement for ionized mixtures. Full article

The incorporation of spore-forming probiotics into thermally processed foods represents a promising strategy to expand functional food availability. In this study, probiotic snacks were formulated from oat and rice-bean matrices as delivery vehicles for Heyndrickxia coagulans (formerly Bacillus coagulans) BC4 spores. The effects of baking and microwave processing, packaging, and storage conditions on spore viability and functionality were evaluated. While oven baking (180 °C) preserved viability in both matrices under mild conditions (survival > 90%), dielectric heating induced significant viability loss depending on the matrix. The starch-based rice-bean matrix, characterized by higher post-processing water activity (Aw), suffered a thermal runaway effect, resulting in significant spore inactivation (viability decreased to 6.08 log colony forming units/g (CFU/g); 1.5 min). Conversely, the oat matrix acted as a thermo-physical stabilizer, maintaining high viability (9.41 log CFU/g; 1.5 min) by limiting dielectric energy absorption via its fiber-lipid composition. Oxidative stress and premature germination likely contributed to the viability loss observed in atmospheric packaging during the 30-day storage. The oat matrix mitigated this effect through a dual-protective mode: active radical scavenging (validated by superior ferric reducing ability of plasma (FRAP) values) and passive water binding. Simulated digestion data align with the functional preservation observed, resulting in increased survival for oat-based formulations. Overall, the results demonstrate the feasibility of developing oat and rice-bean snacks enriched with H. coagulans spores and highlight the critical role of matrix and processing conditions in preserving probiotic viability. Full article

Background/Objectives: Liquid profiling of molecular and epigenetic markers in bodily fluids is an expanding field of cancer biomarker research. Recent research activity also reveals the human satellite 2 (HSAT2) repetitive element cell-free DNA (cfDNA) as a potential cancer biomarker. Based on our recent results from targeted sequencing of HSAT2 cfDNA, we tested whether a specific HSAT2 sequence (e.g., 95 bp-HSAT2) shows greater cancer enrichment than 114 bp-SAT2, from which it derives, in patients with colon cancer. Methods: By comparing the ratio of 114 bp-HSAT2 to 95 bp-HSAT2, we investigated the increased cancer enrichment of 95 bp-HSAT2 in cfDNA samples obtained from plasma DNA extraction and a hybridization capture assay, in which HSAT2 sequences were captured from plasma using a biotin-labeled probe, in samples from colon cancer patients (n = 60) and polyp-controls (n = 60), and polyp-free controls (n = 60). Results: A correlation analysis between Ct values from DNA extraction and the hybridization capture assay for both 95 bp- and 114 bp-HSAT2 showed a positive correlation in patients with colon cancer and control subjects, indicating that the hybridization capture assay provides HSAT2 levels comparable to those obtained by DNA extraction. With both approaches, we found a lower 114 bp-HSAT2 to 95 bp-HSAT2 ratio in patients with colon cancer than in the control groups. The median ratio of extracted DNA was 62, 78, and 79 in patients with colon cancer, polyp-controls (p = 0.23), and polyp-free controls (p = 0.067), respectively. Capture assay values were 49, 87, and 64 in patients with colon cancer, polyp controls (p = 0.016), and polyp-free controls (p = 0.19), respectively. Even though statistical significance was not achieved in some comparisons, these results suggest that 95 bp-HSAT2 is more abundant in the blood of patients with colon cancer than 114 bp-HSAT2 in non-malignant patients. Conclusions: To our knowledge, this is the first study to conduct a hybridization capture assay using a biotinylated probe as a feasible approach for targeted enrichment of cfDNA from plasma. Our results confirm the outcomes of our recent article based on targeted sequencing and reveal that some specific HSAT2 sequences may exhibit increased cancer abundance. Full article

The 12 November 2025 G4 geomagnetic storm—the third most intense of solar cycle 25—was triggered by a complex shock-ICME (interplanetary coronal mass ejection) structure as a result of three ICMEs and driven shocks that arrived on 11–12 November. The main enhancement in the interplanetary magnetic field occurred in the sheath region behind the shock driven by the second ICME. The Dst index reached −217 nT (the SYM-H index reached −254 nT) and the maximum Kp index was 9-. To comprehensively analyze the causes of the storm and its complex effects on near-Earth space, we used a multi-instrumental data set, involving data from satellite missions (ACE, SDO, PROBA2), GNSS networks, ionosondes, optical instruments, high-frequency radars (SuperDARN-like), and cosmic ray monitors. The auroral oval expanded equatorward (down to ~35° N in America). We recorded a super equatorial plasma bubble that almost reached the auroral oval boundary. The equatorial anomaly crests intensified, exceeding 175 TECU, and shifted poleward (8–10°). At mid-latitudes, the F2 layer critical frequency exhibited a strong negative disturbance (−50%) during the main phase, followed by an unusually prolonged and intense positive phase (+100%). GPS Precise Point Positioning errors increased to 2–3 m at high latitudes and in regions affected by the equatorial bubble. The event also featured a Forbush decrease and ground-level enhancement (GLE 77 according to the database hosted by the University of Oulu) associated with the X5.1 solar flare. The results underscore the complex chain of processes from solar storm to geomagnetic and ionospheric responses, highlighting the risks to satellite-based navigation and communication systems. Full article