Search Results (55)

Brown algae such as Ascophyllum nodosum (AN) and Fucus vesiculosus (FV) are gaining considerable attention as functional feed additives due to their health-beneficial properties. This study evaluated the antioxidant potential of AN and FV extracts in intestinal epithelial cells and the in vivo model Caenorhabditis elegans (C. elegans). Aqueous AN and FV extracts were characterized for total phenolic content (TPC), antioxidant capacity (TEAC, FRAP), and phlorotannin composition using LC-HRMS/MS. Antioxidant effects were assessed in vitro, measuring AAPH-induced ROS production in Caco-2 and IPEC-J2 cells via H₂DCF-DA, and in vivo, evaluating the effects of paraquat-induced oxidative stress and AN or FV treatment on worm motility, GST-4::GFP reporter expression, and gene expression in C. elegans. FV exhibited higher total phenolic content, antioxidant capacity (TEAC, FRAP), and a broader phlorotannin profile (degree of polymerization [DP] 2–9) than AN (DP 2–7), as determined by LC-HRMS/MS. Both extracts attenuated AAPH-induced oxidative stress in epithelial cells, with FV showing greater efficacy. In C. elegans, pre-treatment with AN and FV significantly mitigated a paraquat-induced motility decline by 22% and 11%, respectively, compared to PQ-stressed controls. Under unstressed conditions, both extracts enhanced nematode healthspan, with significant effects observed at 400 µg/g for AN and starting at 100 µg/g for FV. Gene expression analysis indicated that both extracts modulated antioxidant pathways in unstressed worms. Under oxidative stress, pre-treatment with AN and FV significantly reduced GST-4::GFP expression. In the nematode, AN was more protective under acute stress, whereas FV better supported physiological function in the absence of stressors. These findings demonstrate that AN and FV counteract oxidative stress in intestinal epithelial cells and in C. elegans, highlighting their potential as stress-reducing agents in animal feed. Full article

Systemic lupus erythematosus (SLE) is a chronic inflammatory autoimmune disease characterized by tissue damage in multiple organs caused by autoantibodies and the resulting immune complexes. One possible way for complement system contribution to onset of autoimmune disorder could be realized by the impairment of C1q-mediated apoptotic clearance as part of human homeostasis. The capacity of C1q to bind early apoptotic cells could be decreased or even lost in the presence of anti-C1q antibodies. A monoclonal anti-idiotypic single-chain (scFv) antibody was selected from the phage library Griffin1” to recognize anti-C1q autoantibodies, purified from sera of lupus nephritis patients. Lupus-prone MRL/lpr mice were injected weekly with scFv A1 fragment-binding anti-C1q antibodies. The number of in vitro and ex vivo studies with collected cells, sera, and organs from the treated animals was performed. scFv treatment changed the percentage of different B-, T-, and NK-cell subpopulations as well as plasma cells and plasmablasts in the spleen and bone marrow. An increase in the levels of splenocyte proliferation, anti-C1q antibodies, and the number of plasma cells producing anti-dsDNA and anti-C1q antibodies were also observed in scFv-treated animals. High levels of proteinuria and hematuria combined with unstable levels of IL10 and IFNγ promote the development of severe lupus and shorten the survival of treated MRL/lpr mice. Therapy with the scFv A1 antibody resulted in BCR recognition on the surface of anti-C1q-specific B-cells and had a disease progression effect, enhancing lupus symptoms in the MRL/lpr mouse model of SLE. Full article

Background/Objectives: Esophageal squamous cell carcinoma (ESCC) is a common form of esophageal cancer with a poor prognosis and limited treatment options. Epidermal growth factor receptor (EGFR), an overexpressed oncogenic gene in all ESCC patients, is an attractive target for developing therapies against ESCC. There is an extremely urgent need to develop immunotherapy tools targeting EGFR for the treatment of ESCC. Methods: In this study, we developed human Interleukin-21 (hIL-21)-armed, chimeric-antigen-receptor-modified T (CAR-T) cells targeting EGFR as a new therapeutic approach. The CAR contains a variable domain of the llama heavy chain of heavy-chain antibodies (VHHs), also known as nanobodies (Nbs), as a promising substitute for the commonly used single-chain variable fragment (ScFv) for CAR-T development. Results: We show that nanobody-derived, EGFR-targeting CAR-T cells specifically kill EGFR-positive esophageal cancer cells in vitro and in animal models. Human IL-21 expression in CAR-T cells further improved their expansion and antitumor ability and were observed to secrete more interferon-gamma (IFN-γ), tumor necrosis factor alpha (TNF-α), and Interleukin-2 (IL-2) when co-cultured with ESCC cell lines in vitro. More CD8⁺ CAR-T cells and CD3⁺CD8⁺CD45RO⁺CD62L⁺ central memory T cells were detected in CAR-T cells expressing hIL-21 cells. Notably, hIL-21-expressing CAR-T cells showed superior antitumor activity in vivo in a KYSE-150 xenograft mouse model. Conclusions: Our results show that hIL-21-armed, nanobody-derived, EGFR-specific CAR-T cell therapy is a highly promising option for treating ESCC patients. Full article

Chronic pain affects a significant portion of the population, with fewer than 30% achieving adequate relief from existing treatments. This study describes the humanization methodology and characterization of an effective non-opioid single-chain fragment variable (scFv) biologic that reverses pain-related behaviors, in this case by targeting P2X4. After nerve injury, ATP release activates/upregulates P2X4 receptors (P2X4R) sequestered in late endosomes, triggering a cascade of chronic pain-related events. Nine humanized scFv (hscFv) variants targeting a specific extracellular 13-amino-acid peptide fragment of human P2X4R were generated via CDR grafting. ELISA analysis revealed nanomolar binding affinities, with most humanized molecules exhibiting comparable or superior affinity compared to the original murine antibody. Octet measurements confirmed that the lead, HC3-LC3, exhibited nanomolar binding kinetics (KD = 2.5 × 10⁻⁹ M). In vivo functional validation with P2X4R hscFv reversed nerve injury-induced chronic pain-related behaviors with a single dose (0.4 mg/kg, intraperitoneal) within two weeks. The return to naïve baseline remained durably reduced > 100 days. In independent confirmation, the spared nerve injury (SNI) model was similarly reduced. This constitutes an original method whereby durable reversals of chronic nerve injury pain, anxiety and depression measures are accomplished. Full article

Fumonisins are polyketide-derived mycotoxins posing significant health threats to humans and animals. Among these, fumonisin B₁ (FB₁) is the most prevalent mycotoxin, primarily produced by Fusarium verticillioides, especially in maize and its derived products. Tangeretin, a polymethoxyflavonoid, has been identified as having potential medicinal properties, particularly as an antioxidant. To evaluate the antifungal and anti-mycotoxigenic properties of tangeretin and to elucidate the mechanisms underlying its inhibitory effects, assessments of fungal growth, FB₁ production, conidial germination, and cellulase activity, antioxidant capacity and enzyme activities, transcriptomic analysis and gene deletion experiments were conducted. Consequently, tangeretin significantly curtailed fungal growth and FB₁ production and provided protection against pathogenic infection on corn. It affected genes associated with fungal growth, conidial development, and antioxidant response. Furthermore, tangeretin interfered with the supply of biosynthetic substrate necessary for fumonisin production, particularly impacting pathways involved in alanine metabolism, pyruvate metabolism, fatty acid degradation, and sphingolipid metabolism. Notably, tangeretin downregulated four biosynthetic genes (Fum2, Fum3, Fum10 and Fum11) that are involved in the final steps of fumonisin formation. It likely disrupted the MAPK signaling pathway and targeted a putative NmrA- and HSCARG-like protein Fv_Tan1, which was identified as having positive effects on fungal growth and mycotoxin biosynthesis. This study presents a promising approach for controlling fumonisin contamination in agricultural settings. Full article

Chromium is a major heavy metal pollutant that affects the health of both plants and animals. In this study, sunflower seedlings were treated with K₂Cr₇O₄ containing 50, 100, and 250 mg of pure chromium per kilogram of soil. It was found that the chromium was absorbed by the roots and transported within plant tissues to the stems and leaves. Chromium affected sunflower photosynthesis, seen in increased the Fv/Fm values as the chromium concentration rose. Metagenomic sequencing of rhizosphere microbial communities after treatment with 100 mg/kg pure chromium indicated that the rhizosphere microorganisms were resistant to chromium exposure; chromium was found to promote dopamine secretion and chromium complexation by the microorganisms. In addition, chromium was found to reduce microbial production of N₂O reductase and increase the emission of the greenhouse gas N₂O. In addressing the problem of chromium pollution in sunflower farmland, Bacillus sp. strain C8 was isolated and shown to effectively reduce soil chromium contents and chromium absorption by sunflower, thereby reducing the adverse effects of the metal. Furthermore, a gene associated with chromium resistance, LOC118480906, was identified by transcriptome sequencing of sunflower plants. In conclusion, the findings denonstrate: (1) the effect of chromium exposure on sunflower growth and development of sunflower; (2) the ecological effects of chromium exposure on sunflower farmland; (3) the regulation of soil microbes and the identification of resistance associated genes are effective ways to reduce chromium pollution. Full article

Herpes simplex viruses (HSV-1 and HSV-2), which can be transmitted both orally and sexually, cause lifelong morbidity and in some cases, meningitis and encephalitis. While both the passive transfer of neutralizing antibodies and placental transfer of anti-HSV monoclonal antibodies (Mabs) have shown therapeutic promise in animal models, clinical trials have yet to identify approved immunotherapeutics for herpes infection. Here, we present strategies for the generation of recombinant bispecific antibodies (BsAbs) that target different domains of glycoprotein D (gD), crucial for HSV entry, that have the potential to outperform the effect of individual Mabs to curb herpes infection. Specifically, we selected three pairs of Mabs from our extensive panel for BsAb design and production based on their binding site and ability to block virus entry. Actual binding of BsAbs to gD and epitope availability on gD after BsAb binding were characterized using surface plasmon resonance (SPR) and inhibition by IgG Fab fragments generated from selected Mabs. While one BsAb exhibited an additive effect similar to that observed using a combination of the Mabs utilized for its generation, two showed antagonistic effects, suggesting that the simultaneous engagement of two epitopes or selective binding to one affected their activity against HSV. One BsAb (DL11/1D3) targeting the binding site for both nectin-1 and HVEM receptors demonstrated synergistic inhibitory activity against HSV, outperforming the effect of the individual antibodies. Recombinant DL11/1D3 antibody variants, in which the size of one or both paratopes was decreased to single chains (scFv-Fc), highlighted differences in potency depending on antibody size and format. We propose that BsAbs to individual glycoproteins offer a potential avenue for herpes therapeutics, but their design, mechanism of action, antibody format, and epitope engagement require careful consideration of structure for optimal efficacy. Full article

Chronic stress, a risk factor for many neuropsychiatric conditions, causes dysregulation in the immune system in both humans and animal models. Additionally, inflammation and synapse loss have been associated with deficits in social behavior. The complement system, a key player of innate immunity, has been linked to social behavior impairments caused by chronic stress. However, it is not known whether complement inhibition can help prevent neuroinflammation and behavioral deficits caused by chronic stress. In this study, we investigated the potential of a site-targeted complement inhibitor to ameliorate chronic stress-induced changes in social behavior and inflammatory markers in the prefrontal cortex (PFC) and hippocampus. Specifically, we investigated the use of C2-Crry, which comprises a natural antibody-derived single-chain antibody (ScFv) targeting domain-designated C2, linked to Crry, a C3 activation inhibitor. The C2 targeting domain recognizes danger-associated molecular patterns consisting of a subset of phospholipids that become exposed following cell stress or injury. We found that systemic administration of C2-Crry attenuated chronic stress-induced social behavioral impairments in mice. Furthermore, C2-Crry administration significantly decreased microglia/macrophage and astrocyte activation markers in the PFC and hippocampus. These findings suggest that site-targeted complement inhibition could offer a promising, safe, and effective strategy for treating chronic stress induced behavioral and immune function disorders. Full article

Edible insects have been evaluated as an alternative and sustainable source of protein because of their nutritive and functional properties for humans and domestic animals. The objective of this study was to assess the use of chemometric [principal component analysis (PCA) and partial least squares (PLS)] combined with Rapid Visco Analyser (RVA) profiles to evaluate the addition of cricket powder (CKP) to chickpea (CPF) and flaxseed (FxF) flours. The results of this study showed that the addition of CKP powder to both CPF and FxF flours affects the pasting properties of the samples; in particular, a reduction in the peak (PV) and final viscosity (FV) was observed. The use of chemometric data techniques such as PCA and PLS regression allowed for a better interpretation of the RVA profiles. Both PCA and PLS regression allowed to qualitative and quantitatively identify the addition level of CKP powder to CPF and FxF flour samples. Differences in the PLS loadings associated with the RVA profile due to the addition of cricket powder were observed. The development of these methodologies will provide researchers and the food industry with better tools to both improve and monitor the quality of ingredients with functional properties as well as to further understand the use of insects as alternative sources of protein. Full article

Chimeric antigen receptor (CAR) T cells represent a revolutionary immunotherapy that allows specific tumor recognition by a unique single-chain fragment variable (scFv) derived from monoclonal antibodies (mAbs). scFv selection is consequently a fundamental step for CAR construction, to ensure accurate and effective CAR signaling toward tumor antigen binding. However, conventional in vitro and in vivo biological approaches to compare different scFv-derived CARs are expensive and labor-intensive. With the aim to predict the finest scFv binding before CAR-T cell engineering, we performed artificial intelligence (AI)-guided molecular docking and steered molecular dynamics analysis of different anti-CD30 mAb clones. Virtual computational scFv screening showed comparable results to surface plasmon resonance (SPR) and functional CAR-T cell in vitro and in vivo assays, respectively, in terms of binding capacity and anti-tumor efficacy. The proposed fast and low-cost in silico analysis has the potential to advance the development of novel CAR constructs, with a substantial impact on reducing time, costs, and the need for laboratory animal use. Full article

The cultivation of corn holds immense importance as a foundational global grain crop, catering to human sustenance and serving as vital animal feed. Moreover, corn plays a substantial role in biofuel production. Additionally, cultivating corn can have a positive effect on crop rotation by improving soil quality and reducing erosion. In a pot trial using six distinct compost variations derived from different organic wastes as fertilizers for GS210 corn, specific indices, such as Fv/Fm (0.80, 0.80, 0.81), Fv/F0 (4.07, 3.99, 4.03), PI (4.62, 4.22, 5.21), and RC/ABS (1.71, 1.68, 2.01), exhibited the highest values. Interestingly, mineral fertilization with NPK displayed significant benefits on various growth parameters like plant height (188.9 cm), cob length (17.50 cm), grains per cob (324.0), and thousand-grain weight (MTZ) (285.2). The difference in the cob grain count between NPK mineral fertilization and the control reached 168.5 grains, which was statistically confirmed. Furthermore, the grain’s protein content notably increased with mineral fertilization (9.5) compared to the control (8.5). While organic fertilizers showed lower outcomes (9.1–9.3) than NPK mineral fertilization, they generally outperformed the control (8.5). This prompts the need for future studies to assess the effectiveness of individual organic fertilizers in combination with mineral nitrogen fertilization. Full article

The genus Flavivirus is a group of arthropod-borne single-stranded RNA viruses, which includes important human and animal pathogens such as Japanese encephalitis virus (JEV), Zika virus (ZIKV), Dengue virus (DENV), yellow fever virus (YFV), West Nile virus (WNV), and Tick-borne encephalitis virus (TBEV). Reverse genetics has been a useful tool for understanding biological properties and the pathogenesis of flaviviruses. However, the conventional construction of full-length infectious clones for flavivirus is time-consuming and difficult due to the toxicity of the flavivirus genome to E. coli. Herein, we applied a simple, rapid, and bacterium-free circular polymerase extension reaction (CPER) method to synthesize recombinant flaviviruses in vertebrate cells as well as insect cells. We started with the de novo synthesis of the JEV vaccine strain SA-14-14-2 in Vero cells using CPER, and then modified the CPER method to recover insect-specific flaviviruses (ISFs) in mosquito C6/36 cells. Chimeric Zika virus (ChinZIKV) based on the Chaoyang virus (CYV) backbone and the Culex flavivirus reporter virus expressing green fluorescent protein (CxFV-GFP) were subsequently rescued in C6/36 cells. CPER is a simple method for the rapid generation of flaviviruses and other potential RNA viruses. A CPER-based recovery system for flaviviruses of different host ranges was established, which would facilitate the development of countermeasures against flavivirus outbreaks in the future. Full article

This paper explores the potential of infrared thermography and geostatistics in animal production and presents the results of the application of the combination of these techniques, contributing significantly to efforts to obtain animals’ responses to the environments in which they are located and thereby ensuring improvements in productivity and animal welfare. The objective was to verify the variability in surface temperature in pigs submitted to different climate control systems using geostatistics. Three growing animals per stall were selected. Dry bulb temperature (Tbd, °C), relative humidity (RH, %) and thermal images were recorded at 08:00 and 12:00 h. To analyze the data, semivariograms were made, the theoretical model was validated and kriging maps were constructed. The mean temperature of the pigs in the pen with adiabatic evaporative cooling (AEC) ranged from 32.40 to 36.25 °C; for the pigs in the forced ventilation (FV) pen, the range of variation was from 32.51 to 36.81 °C. In the control group (Con), with natural ventilation, the average temperature was 37.51 to 38.45 °C. The geostatistical analysis provided a mathematical model capable of illustrating the variation in temperature in the caudal–dorsal regions of the pigs according to the environments to which the animals were subjected. Full article

Surface plasmon resonance (SPR) is a very sensitive measure of biomolecular interactions but is generally too expensive for routine analysis of clinical samples. Here we demonstrate the simplified formation of virus-detecting gold nanoparticle (AuNP) assemblies on glass using only aqueous buffers at room temperature. The AuNP assembled on silanized glass and displayed a distinctive absorbance peak due to the localized SPR (LSPR) response of the AuNPs. Next, assembly of a protein engineering scaffold was followed using LSPR and a sensitive neutron reflectometry approach, which measured the formation and structure of the biological layer on the spherical AuNP. Finally, the assembly and function of an artificial flu sensor layer consisting of an in vitro-selected single-chain antibody (scFv)-membrane protein fusion was followed using the LSPR response of AuNPs within glass capillaries. In vitro selection avoids the need for separate animal-derived antibodies and allows for the rapid production of low-cost sensor proteins. This work demonstrates a simple approach to forming oriented arrays of protein sensors on nanostructured surfaces that uses (i) an easily assembled AuNP silane layer, (ii) self-assembly of an oriented protein layer on AuNPs, and (iii) simple highly specific artificial receptor proteins. Full article

The increased use of wireless technology causes a significant exposure increase for all living organisms to radio frequency electromagnetic fields (RF-EMF). This comprises bacteria, animals, and also plants. Unfortunately, our understanding of how RF-EMF influences plants and plant physiology remains inadequate. In this study, we examined the effects of RF-EMF radiation on lettuce plants (Lactuca sativa) in both indoor and outdoor environments using the frequency ranges of 1890–1900 MHz (DECT) at 2.4 GHz and 5 GHz (Wi-Fi). Under greenhouse conditions, RF-EMF exposure had only a minor impact on fast chlorophyll fluorescence kinetics and no effect on plant flowering time. In contrast, lettuce plants exposed to RF-EMF in the field showed a significant and systemic decrease in photosynthetic efficiency and accelerated flowering time compared to the control groups. Gene expression analysis revealed significant down-regulation of two stress-related genes in RF-EMF-exposed plants: violaxanthin de-epoxidase (VDE) and zeaxanthin epoxidase (ZEP). RF-EMF-exposed plants had lower Photosystem II’s maximal photochemical quantum yield (F_V/F_M) and non-photochemical quenching (NPQ) than control plants under light stress conditions. In summary, our results imply that RF-EMF might interfere with plant stress responses and reduced plant stress tolerance. Full article