Search Results (75)

The only vaccine against Haemonchus contortus is limited by short-lived antibody persistence and the need for frequent booster immunizations. This study leveraged the advantages of nano-adjuvants in enhancing antigen presentation and immune regulation to evaluate the efficacy of novel adjuvants (IMX, AddaS03) and the conventional QuilA combined with H11 protein. Goats were divided into four groups (IMX + H11, AddaS03 + H11, QuilA + H11, and infected control). They were immunized three times and challenged with 6000 infective third-stage larvae (iL3s) of H. contortus on the day of the third immunization, with the experiment lasting for 98 days. The results showed that vaccination with IMX + H11 conferred the strongest protection, demonstrating 88.3% efficacy in fecal egg count (FEC) reduction and 75.8% efficacy against worm burden, followed by QuilA + H11 (85.2% FEC reduction and 68% worm burden reduction) and AddaS03 + H11 (79.4% FEC reduction and 61.3% worm burden reduction). Serum IgG analysis revealed high antibody levels in all immunized groups. Cytokine detection found that IMX + H11 significantly upregulated IL-2 and IFN-γ expression in PBMCs and TNF-α expression in splenocytes, activating Th1-type responses and immune memory. QuilA + H11 showed weaker Th1 activation, and AddaS03 + H11 faced limitations due to insufficient antibody persistence for long-term protection. These findings suggest that IMX can induce highly efficient humoral and cellular immunity, providing a new direction for the optimization of H. contortus vaccines and suggesting the importance of nano-adjuvants for precise regulation of immune patterns. Full article

CD44, a pivotal cell surface molecule, plays a crucial role in many cellular functions, including cell-cell interactions, adhesion, and migration. It serves as a receptor for hyaluronic acid and is involved in lymphocyte activation, recirculation, homing, and hematopoiesis. Moreover, CD44 is a commonly used cancer stem cell marker associated with tumor progression and metastasis. The development of CD44 aptamers that specifically target CD44 can be utilized to target CD44-positive cells, including cancer stem cells, and for drug delivery. Building on the primary sequences of our previously selected thioaptamers (TAs) and observed variations, we developed a bead-based X-aptamer (XA) library by conjugating drug-like ligands (X) to the 5-positions of certain uridines on a complete monothioate backbone. From this, we selected an XA with high affinity to the CD44 hyaluronic acid binding domain (HABD) from a large combinatorial X-aptamer library modified with N-acetyl-2,3-dehydro-2-deoxyneuraminic acid (ADDA). This XA demonstrated an enhanced binding affinity for the CD44 protein up to 23-fold. The selected CD44 X-aptamers (both amine form and ADDA form) also showed enhanced binding affinity to CD44-overexpressing human ovarian cancer IGROV cells. Secondary structure predictions of CD44 using MFold identified several binding motifs and smaller constructs of various stem-loop regions. Among our identified binding motifs, X-aptamer motif 3 and motif 5 showed enhanced binding affinity to CD44-overexpressing human ovarian cancer IGROV cells with ADDA form, compared to the binding affinities with amine form and scrambled sequence. The effect of ADDA as a binding affinity enhancer was not uniform within the aptamer, highlighting the importance of optimal ligand positioning. The incorporation of ADDA not only broadened the XA’s chemical diversity but also increased the binding surface area, offering enhanced specificity. Therefore, the strategic use of site-directed modifications allows for fine-tuning aptamer properties and offers a flexible, generalizable framework for developing high-performance aptamers that target a wide range of molecules. Full article

Cyanobacteria and their toxins can have multiple effects on agricultural productivity and water bodies. Cyanotoxins can be transported to nearby crops and fields during irrigation and may pose a risk to animal health through water sources. Spatial and temporal variations in cyanotoxin concentrations have been reported for large freshwater sources such as lakes and reservoirs, but there are fewer studies on smaller agricultural surface water bodies. To determine whether spatiotemporal patterns of the cyanotoxin microcystin occurred in agricultural waters used for crop irrigation and livestock watering, three agricultural ponds on working farms in Georgia, USA, were sampled monthly within a fixed spatial grid over a 17-month period. Microcystin concentrations, which ranged between 0.04 and 743.75 ppb, were determined using microcystin–ADDA ELISA kits. Temporal stability was assessed using mean relative differences between microcystin concentrations at each location and averaged concentrations across ponds on each sampling date. There were locations or zones in all three ponds that were consistently higher or lower than the average daily microcystin concentrations throughout the year, with the highest microcystin concentrations occurring in winter. Additionally, microcystin patterns were strongly correlated with the patterns of chlorophyll, phycocyanin, and turbidity. The results of this work showed that consistent spatiotemporal patterns in cyanotoxins can occur in produce irrigation and livestock watering ponds, and this should be accounted for when developing agricultural water monitoring programs. Full article

The Automated Driving Data Analyzer (ADDA) is a modular application written in Python for the synchronized acquisition and analysis of physiological, vehicle, and interface data. It provides a managed data acquisition process and one-click data analysis. It also provides raw data storage and systematic archiving of data sets. ADDA integrates real-time data from a BeamNG vehicle simulation, Pupil Labs eye-tracking system, hand-tracking, and cardiac data. This integration allows the simultaneous recording and analysis of multiple data streams, which can be visualized and controlled through a graphical user interface (GUI) built with Tkinter. The application is designed to help researchers and engineers analyze driving behavior under different conditions, enabling a deeper understanding of the interactions between the driver and automated driving functions. Full article

Sediment management is fundamental for managing mountain watercourses and their upslope catchment. A multidisciplinary approach—not limited to the discipline of hydraulics—is necessary for investigating the alterations in sediment transport along the watercourse by detecting those reaches dominated by erosion and deposition processes, by quantifying the sediment volume change, by assessing the functionality of the existing torrent control structures, and by delimitating the riparian vegetation patches. To pursue these goals, specific continuous monitoring is essential, despite being extremely rare in mountain catchments. The present study proposed an integrated approach to determine the hydro-morphological–sedimentological–ecological state of a mountain watercourse though field- and desk-based analyses. Such an integral approach includes a rainfall–runoff model, a morphological change analysis and the application of empirical formulations for estimating peak discharge, mobilizable sediment/large wood volume and watercourse hydraulic capacity, at reach and catchment scales. The procedure was tested on the Upper Adda River catchment (North Italy). The results identified where and with what priority maintenance and monitoring activities must be carried out, considering sediment regime, torrent control structures and vegetation. This study is an example of how it is possible to enhance all existing information through successive qualitative and quantitative approximations and to concentrate new resources (human and economic) on specific gaps, for drafting a scientifically robust and practical sediment management plan. Full article

Background: The rapid production of influenza vaccines is crucial to meet increasing pandemic response demands. Here, we developed plant-made vaccines comprising centralized consensus influenza hemagglutinin (HA-con) proteins (H1 and H3 subtypes) conjugated to a modified plant virus, tobacco mosaic virus (TMV) nanoparticle (TMV-HA-con). Methods: We compared immune responses and protective efficacy against historical H1 or H3 influenza A virus infections among TMV-HA-con, HA-con protein combined with AddaVax™ adjuvant, and whole-inactivated virus vaccine (Fluzone^®). Results: Immunogenicity studies demonstrated robust IgG, IgM, and IgA responses in the TMV-HA-con and HA-con protein vaccinated groups, with relatively low induction of interferon (IFN)-γ⁺ T-cell responses across all vaccinated groups. The TMV-HA-con and HA-con protein groups displayed partial protection (100% and 80% survival) with minimal weight loss following challenge with two H1N1 strains. The HA-con protein group exhibited 80% and 100% survival against two H3 strains, whereas the TMV-HA-con groups showed reduced protection (20% survival). The Fluzone^® group conferred 20–100% survival against two H1N1 strains and one H3N1 strain, but did not protect against H3N2 infection. Conclusions: Our findings indicate that TMV-HA and HA-con protein vaccines with adjuvant induce protective immune responses against influenza A virus infections. Furthermore, our results underscore the potential of plant-based production using TMV-like nanoparticles for developing influenza A virus candidate vaccines. Full article

Inducing T lymphocyte (T-cell) activation and proliferation with specificity against a pathogen is crucial in vaccine formulation. Assessing vaccine candidates’ ability to induce T-cell proliferation helps optimize formulation for its safety, immunogenicity, and efficacy. Our in-house vaccine candidates use microparticles (MPs) and nanoparticles (NPs) to enhance antigen stability and target delivery to antigen-presenting cells (APCs), providing improved immunogenicity. Typically, vaccine formulations are screened for safety and immunostimulatory effects using in vitro methods, but extensive animal testing is often required to assess immunogenic responses. We identified the need for a rapid, intermediate screening process to select promising candidates before advancing to expensive and time-consuming in vivo evaluations. In this study, an in vitro overlay assay system was demonstrated as an effective high-throughput preclinical testing method to evaluate the immunogenic properties of early-stage vaccine formulations. The overlay assay’s effectiveness in testing particulate vaccine candidates for immunogenic responses has been evaluated by optimizing the carboxyfluorescein succinimidyl ester (CFSE) T-cell proliferation assay. DCs were overlaid with T-cells, allowing vaccine-stimulated DCs to present antigens to CFSE-stained T-cells. T-cell proliferation was quantified using flow cytometry on days 0, 1, 2, 4, and 6 upon successful antigen presentation. The assay was tested with nanoparticulate vaccine formulations targeting Neisseria gonorrhoeae (CDC F62, FA19, FA1090), measles, H1N1 flu prototype, canine coronavirus, and Zika, with adjuvants including Alhydrogel^® (Alum) and AddaVax™. The assay revealed robust T-cell proliferation in the vaccine treatment groups, with variations between bacterial and viral vaccine candidates. A dose-dependent study indicated immune stimulation varied with antigen dose. These findings highlight the assay’s potential to differentiate and quantify effective antigen presentation, providing valuable insights for developing and optimizing vaccine formulations. Full article

Nipah virus (NiV), of the Paramyxoviridae family, causes highly fatal infections in humans and is associated with severe neurological and respiratory diseases. Currently, no commercial vaccine is available for human use. Here, eight structure-based mammalian-expressed recombinant proteins harboring the NiV surface proteins, fusion glycoprotein (F), and the major attachment glycoprotein (G) were produced. Specifically, prefusion NiV-F and/or NiV-G glycoproteins expressed in monomeric, multimeric (trimeric F and tetra G), or chimeric forms were evaluated for their properties as sub-unit vaccine candidates. The antigenicity of the recombinant NiV glycoproteins was evaluated in intramuscularly immunized mice, and the antibodies in serum were assessed. Predictably, all homologous immunizations exhibited immunogenicity, and neutralizing antibodies to VSV-luciferase-based pseudovirus expressing NiV-GF glycoproteins were found in all groups. Comparatively, neutralizing antibodies were highest in vaccines designed in their multimeric structures and administered as bivalent (GMYtet + GBDtet) and trivalent (Ftri + GMYtet + GBDtet). Additionally, while all adjuvants were able to elicit an immunogenic response in vaccinated groups, bivalent (GMYtet + GBDtet) and trivalent (Ftri + GMYtet + GBDtet) induced more potent neutralizing antibodies when administered with oil-in-water nano-emulsion adjuvant, AddaS03. For all experiments, the bivalent GMYtet + GBDtet was the most immunogenic vaccine candidate. Results from this study highlight the potential use of these mammalian-expressed recombinant NiV as vaccine candidates, deserving further exploration. Full article

Respiratory syncytial virus (RSV) is a leading cause of severe lower respiratory tract disease of infants and older people. There is an urgent need for safe and effective vaccines against RSV infection. In this study, we analyzed the effects of the immune response and protection with the RSV recombinant G protein extracellular domain (G^ecto) combined with various adjuvants as novel subunit vaccines in mice. All groups receiving RSV G^ecto combined with adjuvants exhibited robust humoral and cellular immunity compared to those receiving an adjuvant alone or inactivated RSV vaccine. The greatest effect was observed in mice receiving G^ecto combined with a CpG ODN + Alum salt adjuvant, resulting in the highest production of neutralizing antibodies against both RSV A and B subtypes, G-specific IgG and IFN-γ production in splenocytes, and interleukin-2 and interferon-γ expression in CD4⁺ T cells. Significant humoral and cellular immune responses were observed in mice immunized with G^ecto combined with AddaS03™ or cyclosporin A adjuvants. The vaccine containing the AddaS03™ adjuvant showed significantly high expression of interleukin-4 in CD4⁺ T cells. Cross-protection against a challenge with either RSV A or B subtypes was observed in the G^ecto plus adjuvant groups, resulting in a significant decrease in viral load and reduced pathological damage in the mouse lungs. These findings offer valuable insights into the development and application of recombinant RSV G-subunit vaccines with adjuvants. Full article

As the clinical application of antibiotics for bacterial skin infections in companion animals becomes increasingly prevalent, the issue of bacterial resistance has become more pronounced. Antimicrobial peptides, as a novel alternative to traditional antibiotics, have garnered widespread attention. In our study, synthetic peptides ADD-A and CBD3-ABU were tested against Staphylococcus pseudintermedius skin infections in KM mice. ADD-A was applied topically and through intraperitoneal injection, compared with control groups and treatments including CBD3-ABU, ampicillin sodium, and saline. Wound contraction, bacterial counts and histology were assessed on days 3 and 11 post-infection. ADD-A and ampicillin treatments significantly outperformed saline in wound healing (p < 0.0001 and p < 0.001, respectively). ADD-A also showed a markedly lower bacterial count than ampicillin (p < 0.0001). Histologically, ADD-A-applied wounds had better epidermal continuity and a thicker epidermis than normal, with restored follicles and sebaceous glands. ADD-A’s effectiveness suggests it as a potential alternative to antibiotics for treating skin infections in animals. Full article

MF59 and AS03 are squalene emulsion-based vaccine adjuvants with similar compositions and droplet sizes. Despite their broad use in licensed influenza vaccines, few studies compared their adjuvant effects and action mechanisms side by side. Considering the majority of adjuvants act on dendritic cells (DCs) to achieve their adjuvant effects, this study compared AddaVax and AddaS03 with similar compositions to MF59 and AS03 adjuvants to enhance antigen uptake, DC maturation, ovalbumin (OVA), and seasonal influenza vaccine-induced immune responses. Considering MF59 was reported to activate MyD88 to mediate its adjuvant effects, this study also investigated whether the above-explored adjuvant effects of AddaVax and AddaS03 depended on MyD88. We found AddaVax more potently enhanced antigen uptake at the local injection site, while AddaS03 more potently enhanced antigen uptake in the draining lymph nodes. AddaS03 but not AddaVax stimulated DC maturation. Adjuvant-enhanced antigen uptake was MyD88 independent, while AddaS03-induced DC maturation was MyD88 dependent. AddaVax and AddaS03 similarly enhanced OVA-induced IgG and subtype IgG1 antibody responses as well as influenza vaccine-induced hemagglutination inhibition antibody titers, while AddaS03 more potently enhanced OVA-specific IgG2c antibody responses. Both adjuvants depended on MyD88 to enhance vaccine-induced antibody responses, while AddaVax depended more on MyD88 to achieve its adjuvant effects. Our study reveals similarities and differences of the two squalene emulsion-based vaccine adjuvants, contributing to our improved understanding of their action mechanisms. Full article

It is clear that new approaches are needed to promote broadly protective immunity to viral pathogens, particularly those that are prone to mutation and escape from antibody-mediated immunity. CD4+ T cells, known to target many viral proteins and highly conserved peptide epitopes, can contribute greatly to protective immunity through multiple mechanisms. Despite this potential, CD4+ T cells are often poorly recruited by current vaccine strategies. Here, we have analyzed a promising new adjuvant (R-DOTAP), as well as conventional adjuvant systems AddaVax with or without an added TLR9 agonist CpG, to promote CD4+ T cell responses to the licensed vaccine Flublok containing H1, H3, and HA-B proteins. Our studies, using a preclinical mouse model of vaccination, revealed that the addition of R-DOTAP to Flublok dramatically enhances the magnitude and functionality of CD4+ T cells specific for HA-derived CD4+ T cell epitopes, far outperforming conventional adjuvant systems based on cytokine EliSpot assays and multiparameter flow cytometry. The elicited CD4+ T cells specific for HA-derived epitopes produce IL-2, IFN-γ, IL-4/5, and granzyme B and have multifunctional potential. Hence, R-DOTAP, which has been verified safe by human studies, can offer exciting opportunities as an immune stimulant for next-generation prophylactic recombinant protein-based vaccines. Full article

Existing mRNA COVID-19 vaccines have shown efficacy in reducing severe cases and fatalities. However, their effectiveness against infection caused by emerging SARS-CoV-2 variants has waned considerably, necessitating the development of variant vaccines. Ideally, next-generation vaccines will be capable of eliciting broader and more sustained immune responses to effectively counteract new variants. Additionally, in vitro assays that more closely represent virus neutralization in humans would greatly assist in the analysis of protective vaccine-induced antibody responses. Here, we present findings from a SARS-CoV-2 VLP vaccine encompassing three key structural proteins: Spike (S), Envelope (E), and Membrane (M). The VLP vaccine effectively produced neutralizing antibodies as determined by surrogate virus neutralization test, and induced virus-specific T-cell responses: predominantly CD4⁺, although CD8⁺ T cell responses were detected. T cell responses were more prominent with vaccine delivered with AddaVax compared to vaccine alone. The adjuvanted vaccine was completely protective against live virus challenge in mice. Furthermore, we utilized air–liquid-interface (ALI)-differentiated human nasal epithelium (HNE) as an in vitro system, which authentically models human SARS-CoV-2 infection and neutralization. We show that immune sera from VLP-vaccinated mice completely neutralized SARS-CoV-2 virus infection, demonstrating the potential of ALI-HNE to assess vaccine induced Nab. Full article

Studying domain adaptation is a recent research trend. Generally, many generative models that researchers have studied perform well on training data from a specific domain. However, their ability to be generalized to other domains might be limited. Therefore, a growing body of research has utilized domain adaptation techniques to address the problem of generative models being vulnerable to input from other domains. In this paper, we focused on generative models and representation learning. Generative models have received a lot of attention for their ability to generate various types of data such as images, music, and text. In particular, studies utilizing generative adversarial neural networks (GANs) and autoencoder structures have received a lot of attention. In this paper, we solved the domain adaptation problem by reconstructing real image data using an autoencoder structure. In particular, reconstructed image data, considered a type of noisy image data, are used as input data. How to reconstruct data by extracting features and selectively transforming them in order to reduce differences in characteristics between domains entails representative learning. Considering these research trends, this paper proposed a novel methodology combining bidirectional feature learning and generative networks to innovatively approach the domain adaptation problem. It could improve the adaptation ability by accurately simulating the real data distribution. The experimental results show that the proposed model outperforms the traditional DANN and ADDA. This demonstrates that combining bidirectional feature learning and generative networks is an effective solution in the field of domain adaptation. These results break new ground in the field of domain adaptation. They are expected to provide great inspiration for future research and applications. Finally, through various experiments and evaluations, we verify that the proposed approach outperforms the existing works. We conducted experiments for representative generative models and domain adaptation techniques and found that the proposed approach was effective in improving data and domain robustness. We hope to contribute to the development of domain-adaptive models that are robust to the domain. Full article

We have studied the interference caused by amplitude and phase distortions induced by rain in ultra-wideband communication systems designed for using amplitude modulation in GeoSurf future satellite constellations. The results concern radio links simulated with the synthetic storm technique at Spino d’Adda (Italy), Madrid (Spain) and Tampa (Florida), which are sites located in different climatic regions. The conclusions are (a) the three sites, although in different climatic zones, are practically indistinguishable; (b) the channel signal-to-noise ratio can be increased or decreased by interference with equal probability. Channel theoretical capacity loss, even in the worst case, is very limited and rain, therefore, does not cause significant linear distortions in ultra-wideband channels at millimeter waves; therefore, these channels could be used at millimeter waves. Full article