Search Results (1,123)

The vast majority of viruses causing human and animal diseases are enveloped—their virions contain an outer lipid bilayer originating from a host cell. Small molecule antivirals targeting the lipid bilayer cover the broadest spectrum of viruses. In this context, we consider the chemical nature and mechanisms of action of membrane-targeting antivirals. They can affect virions by (1) physically modulating membrane properties to inhibit fusion of the viral envelope with the cell membrane, (2) physically affecting envelope lipids and proteins leading to membrane damage, pore formation and lysis, (3) causing photochemical damage of unsaturated membrane lipids resulting in integrity loss and fusion arrest. Other membrane-active compounds can target host cell membranes involved in virion’s maturation, coating, and egress (endoplasmic reticulum, Golgi apparatus, and outer membrane) affecting these last stages of viral reproduction. Both virion- and host-targeting membrane-active molecules are promising concepts for broad-spectrum antivirals. A panel of approved antivirals would be a superior weapon to respond to and control emerging disease outbreaks caused by new viral strains and variants. Full article

Background/Objectives: Liquid biopsy is a minimally invasive alternative to tissue biopsy and is used to obtain information about a disease from a blood sample or other body fluids. In the context of cancer, circulating tumor cells (CTC) can be used as biomarkers to determine the nature of the tumor, its stage of progression, and the efficiency of the administered therapy through monitoring. However, the low concentration of CTCs in blood (1–10 cells/mL) is a challenge for their isolation. Therefore, a minimally invasive medical device (BMProbe™) was developed that isolates CTCs via antigen–antibody binding directly from the bloodstream. Current investigations focus on the process of detaching bound cells from the BMProbe™ surface for cell cultivation and subsequent drug testing to enable personalized therapy planning. Methods: This article presents two approaches for detaching LNCaP cells from anti-EpCAM coated BMProbes™: enzymatic detachment using TrypLE™ and detachment through enzymatic pretreatment with supplementary flow-induced shear stress. The additional shear stress is intended to increase the detachment efficiency. To determine the flow rate required to gently detach the cells, a computational fluid dynamics (CFD) simulation was carried out. Results: The experimental test results demonstrate that 91% of the bound cells can be detached enzymatically within 10 min. Based on the simulation, a maximum flow rate of 47.76 mL/min was defined in the flow detachment system, causing an average shear stress of 8.4 Pa at the probe edges. The additional flow treatment did not increase the CTC detachment efficiency. Conclusions: It is feasible that the detachment efficiency can be further increased by a longer enzymatic incubation time or higher shear stress. The influence on the integrity and viability of cells must, however, be considered. Full article

This research analyzes the innovative development of carnauba wax coatings enriched with essential oils (EOs: lemon, orange, grapefruit, clove, oregano, and cinnamon) or fruit by-products (FBPs: avocado, tomato, carrot, orange, lemon, and grapefruit) to improve postharvest preservation of organic oranges and lemons. Six EOs and six FBPs were evaluated for total phenolic content (TPC) and in vitro antifungal activity against Penicillium digitatum. Based on results, grapefruit, oregano, and clove EOs were selected for lemons, while avocado, orange, and grapefruit FBPs were selected for oranges. An in vivo test at 20 °C for 15 days with carnauba wax coatings assessed antifungal performance. Clove EO and avocado FBP showed strong in vitro inhibition and consistent hyphal suppression (~100 and ~82%, respectively). In vivo, coatings with grapefruit EO and avocado FBP significantly reduced fungal decay and sporulation (~75%) in lemons and oranges, respectively. Coated fruits also retained weight losses by ~25% compared to uncoated ones. These findings suggest that phenolic-rich natural extracts, especially from agro-industrial residues like avocado peels, offer a promising and sustainable strategy for postharvest citrus disease control. Further studies should test coating effectiveness in large-scale trials under refrigeration combined with other preservation strategies. Full article

Salmonella enterica serovar Typhimurium (S. Typhimurium) is an intracellular pathogen capable of surviving and replicating within macrophages, which causes foodborne diseases such as gastroenteritis. To develop a strategy against intracellular bacteria in macrophages, we designed silver-loaded biodegradable polymer nanoparticles functionalized with S. Typhimurium membrane vesicles (MVs). Silver nanoparticles (Ag NPs) were initially encapsulated within biodegradable poly(lactic-co-glycolic) nanoparticles (Ag-P NPs), which were then surface-modified with polyethyleneimine to form Ag-PP NPs. These were subsequently fused with S. Typhimurium MVs via a sonication method to generate Ag-PP@MV NPs. The resulting MV-coated nanoparticles displayed a similar protein profile to that of native MVs and exhibited antibacterial activity against intracellular S. Typhimurium. Notably, the enhanced cellular uptake of the MV-modified NPs contributed to their intracellular bactericidal efficacy. This study highlights MV modification as a promising strategy to improve NP delivery to macrophages for treating persistent intracellular infections. Full article

Oxidized phospholipids (OxPLs) are increasingly recognized as biologically active lipids involved in various pathologies. Both exposure to pathogenic factors and the efficacy of protective mechanisms are critical to disease development. In this study, we characterized an immunoassay that quantified the total capacity of the plasma to degrade or mask OxPLs, thereby preventing their interaction with cells and soluble proteins. OxLDL-coated plates were first incubated with human blood plasma or a control vehicle, followed by an ELISA using a monoclonal antibody specific to oxidized phosphatidylethanolamine. Pretreatment with the diluted blood plasma markedly inhibited mAb binding. The masking assay was optimized by evaluating the buffer composition, the compatibility with various anticoagulants, potential interfering compounds, the kinetic parameters, pre-analytical stability, statistical robustness, and intra- and inter-individual variability. We propose that this masking assay provides a simple immunological approach to assessing protective mechanisms against lipid peroxidation products. Establishing this robust and reproducible method is essential for conducting clinical association studies that explore masking activity as a potential biomarker of the predisposition to a broad range of lipid-peroxidation-related diseases. Full article

Background/Objectives: Beak and feather disease virus (BFDV) is the causative agent of psittacine beak and feather disease (PBFD), affecting psittacine birds. There is currently no commercial vaccine or treatment for this disease. This study developed a novel BFDV coat protein mRNA vaccine encapsidated by TMV coat protein to form pseudovirions (PsVs) and tested its immunogenicity alongside BFDV coat protein (CP) subunit and DNA vaccine candidates. Methods: mRNA and BFDV CP subunit vaccine candidates were produced in Nicotiana benthamiana and subsequently purified using PEG precipitation and gradient ultracentrifugation, respectively. The DNA vaccine candidate was produced in E. coli cells harbouring a plasmid with a BFDV1.1mer pseudogenome. Immunogenicity of the vaccine candidates was evaluated in African grey parrot chicks. Results: Successful purification of TMV PsVs harbouring the mRNA vaccine, and of the BFDV-CP subunit vaccine, was confirmed by SDS-PAGE and western blot analysis. TEM analyses confirmed formation of TMV PsVs, while RT-PCR and RT-qPCR cDNA amplification confirmed encapsidation of the mRNA vaccine candidate within TMV particles. Restriction digests verified presence of the BFDV1.1mer genome in the plasmid. Four groups of 5 ten-week-old African grey parrot (Psittacus erithacus) chicks were vaccinated and received two boost vaccinations 2 weeks apart. Blood samples were collected from all four groups on day 14, 28 and 42, and sera were analysed using indirect ELISA, which showed that all vaccine candidates successfully elicited specific anti-BFDV-CP immune responses. The subunit vaccine candidate showed the strongest immune response, indicated by higher binding titres (>6400), followed by the mRNA and DNA vaccine candidates. Conclusions: The candidate vaccines present an important milestone in the search for a protective vaccine against PBFD, and their inexpensive manufacture could considerably aid commercial vaccine development. Full article

Background/Objectives: The formation of oral biofilms in periodontal pockets and around dental implants with induction of periodontitis or peri-implantitis is an increasing problem in dental health. The intelligent design of a biofilm makes the bacteria embedded in the biofilm matrix highly tolerant to antiseptic therapy, often resulting in tooth or implant loss. The question therefore arises as to which mouthwashes have eradication potential against oral biofilm. Methods: A human oral biofilm model was developed based on donated blood plasma combined with buffy coats, inoculated with oral pathogenic bacterial species found in periodontal disease (Actinomyces naeslundii, Fusobacterium nucleatum, Streptococcus mitis, and Porphyromonas gingivalis). Over a span of 7 days, we tested different mouth rinsing and antiseptic solutions (Chlorhexidine, Listerine^®, NaOCl, Octenisept^®, and Octenident^®) covering the matured biofilm with 24 h renewal. Phosphate-buffered saline (PBS) was used as a control. Bacterial growth patterns were detected via quantitative polymerase chain reaction (qPCR) after 2, 4, and 7 days of treatment. Results: While all groups showed initial bacterial reduction, the control group demonstrated strong regrowth from day 2 to 4. Listerine showed a near-significant trend toward bacterial suppression. Additionally, strain-specific efficacy was observed, with Octenisept^® being most effective against Streptococcus mitis, Octenident^® and NaOCl showing superior suppression of Actinomyces naeslundii, and Listerine^® outperforming other solutions in reducing Fusobacterium nucleatum. Donor-specific, individual variability further influenced treatment outcomes, with distinct trends in bacterial suppression and regrowth observed across donors. Conclusions: These findings underscore the complexity of biofilm-associated infections and highlight the importance of targeted therapeutic approaches for managing bacterial biofilms. In this experiment, the donor-specific outcomes of the antimicrobial effects of the solutions may indicate that genetic predisposition/tolerance to oral infections appears to play a critical role in the control of oral biofilms. Full article

Background: Poststroke dysphagia is a common condition that can lead to complications such as aspiration pneumonia and malnutrition, significantly affecting the quality of life. Most patients recover their swallowing function spontaneously, but in others difficulties persist beyond six months. Can we predict this in advance? On the other hand, there have been recent attempts to use machine learning to predict disease prognosis. Therefore, this study aims to investigate whether machine learning can predict the long-term prognosis for poststroke dysphagia using early videofluoroscopic swallowing study (VFSS) data. Methods: Data from VFSSs performed within 1 month of onset and swallowing status at 6 months were collected retrospectively in patients with dysphagia who experienced their first acute stroke at a university hospital. We selected 14 factors (lip closure, bolus formation, mastication, apraxia, tongue-to-palate contact, premature bolus loss, oral transit time, triggering of pharyngeal swallow, vallecular residue, laryngeal elevation, pyriform sinus residue, coating of the pharyngeal wall, pharyngeal transit time, and aspiration) from the VFSS data, scored them, and analyzed whether they could predict the long-term prognosis using five machine learning algorithms: Random forest, CatBoost classifier, K-neighbor classifier, Light gradient boosting machine, Extreme gradient boosting. These algorithms were combined through an ensemble method to create the final model. Results: In total, we collected data from 448 patients, of which 70% were used for training and 30% for testing. The final model was evaluated using accuracy, precision, recall, F1-score, and Area Under the Receiver Operating Characteristic Curve (AUC), resulting in values of 0.98, 0.94, 0.84, 0.88, and 0.99, respectively. Conclusions: Machine learning models using early VFSS data have shown high accuracy and predictive power in predicting the long-term prognosis of patients with poststroke dysphagia, and they are likely to provide useful information for clinicians. Full article

Fruit and vegetable production is often impacted by microbial pathogens that compromise the quality of produce and lead to significant economic losses at the postharvest stages. Due to their efficacy, agrochemicals are widely applied in disease management; nevertheless, this practice has led to the appearance of microbial strains resistant to these types of agrochemicals. Additionally, there is growing concern among consumers about the presence of these chemical residues in fruits and the negative impacts they cause on multiple ecosystems. In response, there is a growing need for safe, effective, green, and sustainable disease control technologies. Bionanocomposites, with their unique ability to combine nanomaterials and biopolymers that have attractive properties, represents a promising alternative for postharvest disease control. These technologies allow for the development of functional coatings and films with antimicrobial, antioxidant, and barrier properties, which are critical for extending shelf life and preserving fruit quality. Recent advances have demonstrated that integrating nanoparticles, such as ZnO, TiO₂, Ag, and chitosan-based nanosystems, into biopolymeric matrices, like alginate, pectin, starch, or cellulose, can enhance mechanical strength, regulate gas exchange, and control the release of active agents. This review presents systematized information that is focused on the creation of coatings and films based on bionanocomposites for the management of disease in fruits and vegetables. It also discusses the use of diverse biopolymers and nanomaterials and their impact on the quality and shelf life of fruits and vegetables. Full article

In this paper, we present the development of a new semi-distributed interferometer (SDI) biosensor with a Zn, Cu, and Co metal oxide nanopowder coating for the detection of a kidney disease biomarker as a model system. The combination of nanopowder coating with the SDI platform opens up unique opportunities for improving measurement reproducibility while maintaining high sensitivity. The fabrication of sensors is simple, which involves one splice and subsequent cutting at the end of an optical fiber. To ensure specific detection of the biomarker, a monoclonal antibody was immobilized on the surface of the probe. The biosensor has demonstrated an impressive ability to detect biomarkers in a wide range of concentrations, from 1 aM to 100 nM. The theoretical limit of detection was 126 fM, and the attomolar detection level was experimentally achieved. The sensors have achieved a maximum sensitivity of 190 dB/RIU and operate with improved stability and reduced dispersion. Quantitative analysis revealed that the sensor’s response gradually increases with increasing concentration. The signal varies from 0.05 dB at 1 aM to 0.81 dB at 100 nM, and the linear correlation coefficient was R² = 0.96. The sensor showed excellent specificity and reproducibility, maintaining detection accuracy at about 10⁻⁴ RIU. This opens up new horizons for reliable and highly sensitive biomarker detection, which can be useful for early disease diagnosis and monitoring using a cost-effective and reproducible sensor system. Full article

Glutamate, the primary excitatory neurotransmitter in the central nervous system, plays a pivotal role in synaptic signaling, learning, and memory. Abnormal glutamate levels are implicated in various neurological disorders, including epilepsy, Alzheimer’s disease, and ischemic stroke. Despite the utility of magnetic resonance imaging (MRI) and magnetic resonance spectroscopy (MRS) in diagnosing such conditions, the development of effective glutamate-sensitive contrast agents remains a challenge. In this study, we present ultrasmall, citric acid-coated superparamagnetic iron oxide nanoparticles (CA-SPIONs) as highly selective and sensitive MRS probes for glutamate detection. These 5 nm magnetite CA-SPIONs exhibit a stable dispersion in physiological buffers and undergo aggregation in the presence of glutamate, significantly enhancing the T₂ MRS contrast power. At physiological glutamate levels, the CA-SPIONs yielded a pronounced signal change ratio of nearly 60%, while showing a negligible response to other neurotransmitters such as GABA and dopamine. Computational simulations confirmed the mechanism of glutamate-mediated aggregation and its impact on transversal relaxation rates and relaxivities. The sensitivity and selectivity of CA-SPIONs underscore their potential as eco-friendly, iron-based alternatives for future neurological sensing applications targeting glutamatergic dysfunction. Full article

Tomato leaf curl New Delhi virus (ToLCNDV) is among the most prevalent and widely distributed begomovirus infecting chili pepper (Capsicum annuum) and tomato in the Indian subcontinent. In this study, a guide RNA (gRNA) sequence-CRISPR-Cas9 approach was used to target and cleave two essential coding regions in the begomovirus genome. The gRNAs were designed to target conserved regions of the ToLCNDV replication-associated protein (rep) gene or ORF AC1, and/or the coat protein (cp) gene or AV1 ORF, respectively. Based on an alignment of 346 representative ToLCNDV genome sequences, all predicted single nucleotide polymorphisms off-target sites were identified and eliminated as potential gRNA targets. Based on the remaining genome regions, four candidate gRNAs were designed and used to build gRNA-Cas9 duplexed constructs, e.g., containing two gRNAs cloned in tandem, in different combinations (1–4). Two contained two gRNAs that targeted the coat protein gene (cp; AV1 ORF), while the other two constructs targeted both the cp and replication-associated protein gene (rep; AC1 ORF). These constructs were evaluated for the potential to suppress ToLCNDV infection in Nicotiana benthamiana plants in a transient expression-transfection assay. Among the plants inoculated with the duplexed gRNA construct designed to cleave ToLCNDV-AV1 or AC1-specific nucleotides, the construct designed to target both the cp (293–993 nt) and rep (1561–2324) showed the greatest reduction in virus accumulation, based on real-time quantitative PCR amplification, and attenuated disease symptoms, compared to plants inoculated with the DNA-A component alone or mock-inoculated, e.g., with buffer. The results demonstrate the potential for gRNA-mediated suppression of ToLCNDV infection in plants by targeting at least two viral coding regions, underscoring the great potential of CRISPR-Cas-mediated abatement of begomovirus infection in numerous crop species. Full article

Many dogs suffer from pruritus, which is commonly caused by atopic dermatitis and associated with skin inflammation. The immune system and inflammatory response, and in particular the gut–skin axis, are central to the pathogenesis of atopic dermatitis. Managing atopic dermatitis involves complex, iterative treatment plans; early strategies supporting gut–skin health are needed to prevent elevated itching from progressing toward a disease condition requiring drug therapy. This double-blind, placebo-controlled, randomized trial evaluated the ability of a novel, indole-rich canine immune health postbiotic (CIHP) to reduce itching and promote a healthy gut microbiome in dogs with subclinical, but elevated itching behavior. Thirty dogs were stratified into two groups based on baseline scratching frequency, receiving either CIHP or placebo as a powder topper for 28 days. Canine itching was evaluated through accelerometer-based tracking and the Pruritus Visual Analog Scale (PVAS) score on Days 0, 7, 14, 21, and 28. Skin and coat health was assessed on Days 0, 14, and 28, and the gut microbiome was sequenced from fecal samples on Days 0 and 28. CIHP reduced scratching by 20% relative to the baseline (p = 0.032) and PVAS score by 27% compared to the placebo (p = 0.02). CIHP improved skin and coat quality compared to the placebo at Day 14 (p = 0.01) and increased Shannon diversity by 4.6% (p = 0.043), shifting gut microbiome composition. These findings validate this postbiotic’s ability to reduce itching in dogs with subclinical, but elevated itching behavior, provide evidence of promoting a healthy gut–skin axis, and suggest potential as an early intervention in the context of pruritic conditions, as well as for broader immune-related benefits. Full article

Grapevine leafroll disease is one of the most devastating diseases in the global viticulture industry. Grapevine leafroll-associated virus 4 is one of the main pathogens causing this disease. In this study, we obtained the complete genome sequences of two Chinese isolates of GLRaV-4 from ‘Baisainie’ and ‘Fantasy Seedless’ grapevines through high-throughput sequencing and overlapping RT-PCR combined with RACE technology. The sequences contain 13,814 and 13,824 nucleotides and code six open reading frames, respectively. Phylogenetic trees based on the coat protein (CP) and heat shock protein 70 (HSP70) genes show that in addition to other GLRaV-4 strains (strains 5, 6, 9, Pr, and Car), the GLRaV-4 strains were divided into two distinct groups. The two isolates obtained in this study were classified into separate branches within GLRaV-4 Group 1. Additionally, we systematically investigated the interactions between the proteins encoded by GLRaV-4 using the yeast two-hybrid and bimolecular fluorescence complementation techniques. We found significant interactions between the GLRaV-4-encoded p23 and HSP70 and CP. This study first reports the complete genomes of two different GLRaV-4 isolates from China and suggests that p23 protein encoded by GLRaV-4 may play an important role in viral pathogenicity due to its interactions with the other two proteins. Full article

Coronary artery disease is the most common cause of mortality worldwide. Percutaneous coronary intervention represents an important method of treatment. Over time, the methods have been refined to improve safety and efficacy. With the development of drug-eluting stents, in-stent restenosis has importantly decreased, but it remains a relevant concern in terms of the need for additional revascularization procedures or recurrent coronary events. Different platforms, polymers, and anti-proliferative agents have been tested, mostly demonstrating non-inferiority when compared. Additional devices, such as drug-coated balloons, bioresorbable scaffold systems, gene-eluting stents and bioadaptor implants have been developed. As none of the aforementioned methods demonstrated considerable superiority over the others, the search for the ideal treatment method continues. Based on currently available data, the ideal treatment method could be a personalized approach combining different revascularization methods. Additional research with subpopulation group studies, different associated diseases or vessels affected, and longer follow-up are required to determine better subgroups of patients that would benefit most from specific treatment methods. Full article