Low Accumulation Research Articles

Recent advances in polymeric and lipid stimuli-responsive nanocarriers for cell-based cancer immunotherapy.

Conventional cancer therapy has major limitations, including non-specificity, unavoidable side effects, low specific tumor accumulation and systemic toxicity. In recent years, more effective and precise treatment methods have been developed, including cell-based immunotherapy. Carriers that can accurately and specifically target cells and equip them to combat cancer cells are particularly important for developing this therapy. As a result, attention has been drawn to smart nanocarriers that can react to specific stimuli. Thus, stimuli-responsive nanocarriers have attracted increasing attention because they can change their physicochemical properties in response to stimulus conditions, such as pH, enzymes, redox agents, hypoxia, light and temperature. This review highlights recent advances in various stimuli-responsive nanocarriers, discussing loading, targeted delivery, cellular uptake, biocompatibility and immunomodulation in cell-based immunotherapy. Finally, future challenges and perspectives regarding the possible clinical translation of nanocarriers are discussed.

Abstract B070: Dissecting EV dynamics in the sequestration of doxorubicin from breast cancer cells

Abstract Background: Extracellular vesicles (EVs) are one of the three main analytes sought for in liquid biopsy development. EVs are released by all cell types including cancer cells and play a crucial role in cell-cell communication. Breast cancer cells have been shown to utilize EVs as a mechanism to sequester chemotherapeutic agents, thereby conferring resistance to treatment. This study investigates the role of EVs in doxorubicin (DOX) resistance in MDA-MB-231 breast cancer cells. Method: MDA-MB-231 cells were exposed to increasing dose of anthracycline for 6 and 12 months. Resistance was confirmed by cytotoxicity assay and western blot. Resistant and parent cells were labeled for immunohistochemistry and their EVs were isolated via ultracentrifugation. EVs were analysed using nanoscale flow cytometry and sent for mass spectrophotometry. Result: Using nanoscale flow cytometry, we optimized a method to detect DOX-carrying EVs based on the intrinsic fluorescence of the drug. We found that DOX-resistant cells exhibited lower intracellular DOX accumulation and higher expression of the EV marker CD63 compared to their parental counterparts. Conclusion: These results suggest that chemoresistant breast cancer cells may alter their EV production machinery to evade the lethal effects of DOX. Understanding this mechanism could lead to novel strategies to overcome chemoresistance in breast cancer. Citation Format: Sina Halvaei, Jing Xu, Suganthi Chittaranjan, Sharon Gorski, Karla Williams. Dissecting EV dynamics in the sequestration of doxorubicin from breast cancer cells [abstract]. In: Proceedings of the AACR Special Conference: Liquid Biopsy: From Discovery to Clinical Implementation; 2024 Nov 13-16; San Diego, CA. Philadelphia (PA): AACR; Clin Cancer Res 2024;30(21_Suppl):Abstract nr B070.

Abstract 4136511: Targeting ATP Citrate Lyase: A Novel Therapeutic Strategy for Vascular Remodeling in Pulmonary Hypertension

Introduction: Pulmonary arterial hypertension (PAH) is a vascular remodeling disease characterized by a progressive obliteration of distal pulmonary arteries (PA), which is driven by a cancer-like pro-proliferative phenotype of smooth muscle cells (SMCs). A metabolic shift towards glycolysis and changes in the epigenetic landscape contribute to this abnormal phenotype. ATP Citrate Lyase (ACLY) is an enzyme that plays a role in promoting the Warburg effect, lipid synthesis, and chromatin remodeling in cancer. However, its role in PAH remains unknown. We hypothesized that ACLY is upregulated in PAH and supports the abnormal phenotype of PAH-PASMCs . Methods/Results: To investigate this hypothesis, we measured ACLY expression in human distal pulmonary arteries and isolated PASMCs from PAH patients (immunoblot/Immunofluorescence). We found increased expression and activation of ACLY in PAH-PASMCs, with preferential localization in the nucleus. Inhibition of ACLY using BMS-303141 or siACLY decreased PAH-PASMCs proliferation (Ki67/MCM2, PLK1, PCNA) and survival (Annexin V/SURVIVIN). ACLY inhibition also reduced glycolysis markers and increased mitochondrial respiration. Additionally, the inhibition of ACLY lowered cholesterol levels and lipid droplet accumulation in PAH-PASMCs. Further analysis showed that ACLY promotes nuclear acetyl-CoA production, leading to acetylation of specific histone proteins and GCN5-mediated transcriptional activation of genes involved in cell cycle progression. The transcription factor FOXM1 appeared to mediate this genetic signature. In vivo , pharmacological inhibition of ACLY using BMS-303141 significantly improved pulmonary vascular remodeling (Elastica Van Gieson staining, EVG) and right ventricular function (RVSP, mPAP, TAPSE, S-Wave, SV by echocardiography and right heart catheterization) in Su/Hx-challenged rats with established PAH. SMC-specific Acly K.O mice were protected against Su/Hx-induced PAH. Consistently, tamoxifen-induced SMC-specific Acly deletion improved hemodynamics parameters in Su/Hx mice. These therapeutic effects were associated with a reduced acetylation of H3K27 and H3K9 (IF). Ex vivo , inhibition of ACLY attenuated vascular remodeling (EVG) in human precision cut lung slices exposed to a growth factor cocktail. Conclusion: Our study reveals that ACLY plays a critical role in vascular remodeling in PAH and its pharmacological inhibition may represent a novel and attractive avenue as therapeutic treatment.

Changes in total soluble solids concentration, fruit acidity, and yeast assimilable nitrogen in response to altered leaf area to fruit weight ratio in Pinot noir

The increasing consumer demand for lower alcohol wine and the need to mitigate against a warming climate presents new challenges for winegrowers and the desire to produce grapes with lower total soluble solids (sugar) or earlier harvesting, while preserving other wine compounds and attributes, particularly for varieties intended for red wine production. We investigated how reducing vine leaf area through shoot trimming and leaf removal, applied at different severities and timings to modify the leaf area to fruit weight (LA) ratio, affects fruit composition for producing lower alcohol quality Pinot noir wine. Shoot trimming treatments (half canopy, H in 2015/16 and 2016/17, and quarter canopy, Q in 2016/17 by alternately removing leaves after trimming shoots to half) were applied shortly before veraison (V-, E-L 34), during veraison (V, E-L 35) and post-veraison (V+, E-L 36) in three different vineyard locations (Marlborough, Canterbury and Central Otago, New Zealand). Untrimmed vines served as the control. Lateral shoots were removed during treatment application, and regrowth was removed to maintain a consistent leaf area. Results showed that reducing the LA:FW ratio delayed the accumulation and concentration of total soluble solids (reduction of 1.0 to 2.7 °Brix) at harvest in all trimmed vines over both seasons. Berry weight, malic acid concentration, titratable acidity, and pH at harvest were unaffected by trimming. At target TSS levels of 18 °Brix (10 % ethanol, v/v) in the Marlborough vineyard and 20 °Brix (11 % v/v) in the Central Otago vineyard, V+ vines showed malic acid and titratable acidity levels comparable to the control. At the Canterbury vineyard, these parameters remained similar across all treatments at 16 °Brix (8.9 % v/v). Yeast-assimilable nitrogen concentration increased (188 to 411 mg/L) in early trimmed vines. At Central Otago, roots showed lower carbohydrate reserves across all trimming treatments, likely due to the high yield at the site, while at Canterbury, trimming did not result in significant differences in carbohydrate reserves, likely due to the low yield at the site. At the Marlborough vineyard, vines trimmed early and more severely had less starch in their roots, while HV+ vines maintained similar levels of starch to controls. In conclusion, halving the canopy post-veraison (HV+, LA 0.70 m²/kg as observed in Marlborough) could be considered a viable viticultural option to lower sugar accumulation, thereby reducing potential alcohol content, without affecting titratable acidity and pH. This practice offers significant potential for adapting to a warming climate and producing lower alcohol Pinot noir wines in a sustainable manner.

Orthogonal Nano‐Engineering (ONE): Modulating Nanotopography and Surface Chemistry of Aluminum Oxide for Superior Antibiofouling and Enhanced Chemical Stability

AbstractDecoupling certain material surface properties can be key to attaining critical property‐activity relationships that underpin their antibiofouling performance. Here, orthogonal nano‐engineering (ONE) is employed to decouple the influences of nanotopography and surface chemistry on surface antibiofouling. Nanotopography and surface chemistry are systematically varied with a two‐step process. Controlled nanotopography is obtained by electrochemical anodization of aluminum, which generated anodic aluminum oxide (AAO) surfaces with cylindrical nanopores (diameters: 15, 25, and 100 nm). To modify surface chemistry while preserving nanotopography, an ultrathin (≈5 nm) yet stable zwitterionic coating of poly(divinylbenzene‐4‐vinylpyridyl sulfobetaine) is deposited on these surfaces using initiated chemical vapor deposition (iCVD). Antibiofouling performance is assessed by quantifying 48‐h biomass formed by gram positive and negative bacteria. The ONE surfaces demonstrated enhanced antibiofouling performance, with small‐pore nanotopography and zwitterionic chemistry each lowered biomass accumulation by tested species, with potential additive effects. The most effective chemistry‐topography combination (ZW‐AAO15) enabled an overall reduction of 91% for Escherichia coli, 76% for Staphylococcus epidermidis, 69% for Listeria monocytogenes, and 67% for Staphylococcus aureus, relative to the uncoated nanosmooth control. Additionally, the composite ZW coating exhibited encouraging anticorrosion properties under both static and turbulent cleaning conditions, vital to antibiofouling applications in healthcare and food industries.

Site-Specific Modification of Native IgGs with Flexible Drug-Load.

Homogeneous, site-specifically conjugated antibodies have shown to result in antibody-drug conjugates (ADCs) with improved efficacy and tolerability compared to stochastically conjugated ADCs. However, precisely controlling the drug load as well as attaching multiple payload moieties to the antibody remains challenging. Here, we demonstrate the simple and direct modification of native IgG-antibodies at the residue glutamine 295 (Q295) without the need for any protein engineering with flexible drug-to-antibody ratios of one or multiple payloads. The conjugation is enabled through short, positively charged lysine containing peptides and native, commercially available microbial transglutaminase. In proof-of-concept studies, HER2-targeting ADCs based on trastuzumab were generated with drug-to-antibody ratios (DARs) of 2 and 4 of the same or different payloads using orthogonal conjugation chemistries. Quantitative biodistribution studies performed with 111In-radiolabeled conjugates showed high tumour uptake and low accumulation of radioactivity in non-targeted tissues. A single dose study of trastuzumab conjugated to the highly potent payload α-Amanitin demonstrated complete and long-lasting tumour remission and was well-tolerated at all dose levels tested.

Exploring genotypic variation and gene expression associated to cadmium accumulation in bread wheat

Cadmium (Cd) contamination poses significant risks to agricultural productivity and human health, particularly through its accumulation in staple crops such as bread wheat (Triticum aestivum L.). This study evaluated Cd accumulation and tolerance among six bread wheat cultivars exposed to six Cd concentrations (0, 2.5, 5, 10, 15, 20, and 25 mg kg−1 soil). Phenotypic assessments and quantitative real-time PCR (qRT-PCR) were conducted to analyze the expression patterns of TaNRAMP and TaZIP genes in various tissues and developmental stages of wheat, which play crucial roles in Cd uptake and transport. Results demonstrated significant variability in Cd accumulation. The Barat cultivar exhibited the lowest accumulation in grain (ranging from 0.21 to 8.8 mg kg−1) and the highest tolerance. In contrast, Kavir and Pishtaz displayed elevated Cd levels in both grain and straw, while Parsi accumulated more Cd in straw at lower concentrations (56.9 mg kg−1 in Cd concentration of 10 mg kg−1 soil). The gene expression analysis revealed that most cultivars showed increased expression of TaNRAMP genes, particularly TaNRAMP2 in Cd concentration of 10 mg kg−1 soil, which facilitates Cd uptake from the soil, and TaZIP genes, such as TaZIP4 and TaZIP7, involved in transporting Cd within the plant. Notably, the expression of TaZIP1 was significantly lower in cultivars with high Cd accumulation, suggesting a potential regulatory mechanism for Cd tolerance. Furthermore, cultivars exhibiting higher Cd levels correlated with increased expression of stress-responsive genes, indicating a broader response to Cd stress. These findings highlight Barat’s potential for bread-making applications due to its low Cd accumulation, while Morvarid and Pishtaz which show reduced Cd content in the straw even under high Cd exposure are better suited for animal feed. This research underscores the genetic variability of wheat cultivars in response to Cd stress and provides essential insights into the molecular mechanisms underlying Cd accumulation, offering valuable information for breeding programs aimed at developing Cd-tolerant varieties to ensure food security in contaminated regions.

Improvement of RBD-FC Immunogenicity by Using Alum-Sodium Alginate Adjuvant Against SARS-COV-2.

Adjuvants use several mechanisms to boost immunogenicity and to modulate immune response. The strength of adsorption of antigen by adjuvants can be a determinant factor for significant improvement of immunopotentiation. We expressed recombinant RBD-FC in PichiaPink Strain 4 and examined the vaccination of mice by vaccine formulation with different adjuvants (sodium alginate and aluminum hydroxide, alone and together). Sodium alginate significantly increased the immunogenicity and stability of RBD-FC antigen, so RBD-FC formulated with combined alginate and alum (AlSa) and sodium alginate alone showed higher antibody titer and stability. Immunogenicity of RBD-FC:AlSa was determined by serological assays including direct enzyme-linked immunosorbent assay (ELISA) and surrogate virus neutralization test (sVNT). High levels of IgGs and neutralizing antibodies were measured in serum of mice immunized with the RBD-FC:AlSa formulation. On the other hand, cytokines IL-10 and INF-γ were severely accumulated in response to RBD-FC:AlSa, and after 10 days, their accumulation was significantly declined, whereas IL-4 showed the highest and the lowest accumulation in response to alum and alginate, respectively. Our data may suggest that combination of alum and sodium alginate has a better compatibility with RBD-FC in vaccine formulation.

Identification, functional characterization and expression profiling of three triterpene synthases from the legume plant Vigna unguiculata

Oxidosqualene cyclases (OSCs) are important regulatory enzymes involved in cyclization reactions of 2, 3-oxidosqualene to form triterpenes and sterols. This study presents the identification and characterization of three OSC genes, a β – amyrin synthase (VuβAS), a lupeol synthase (VuLUS) and a cycloartenol synthase (VuCAS) in Vigna unguiculata, an edible leguminous plant with high nutritional and nutraceutical value. Phylogenetic analysis showed that the VuβAS, VuLUS and VuCAS were clustered within the clades of previously characterized β – amyrin synthases, lupeol synthases and cycloartenol synthases. Heterologous expression in Saccharomyces cerevisiae and Gas Chromatography – Mass Spectrometry (GC – MS) analysis in different plant stages confirmed their specific functions. VuβAS showed higher expression in roots from early germinating seedlings to older plants (4-day to 28-day), while VuLUS expression levels were higher in the roots of older plants only (14-day to 28-day). VuCAS expression was increased in all the tissues of 4-day seedlings, with a peak in stem and leaves and a lower accumulation in radicles. These findings revealed the presence and function of OSC genes in V. unguiculata, and future research could lead to the discovery of promising biologically active compounds.

LkERF6 enhances drought and salt tolerance in transgenic tobacco by regulating ROS homeostasis

The transcription factor Ethylene Responsive Factor (ERF) is crucial for responding to various environmental stressors. Proteins containing the ERF-associated amphiphilic repression (EAR) motif often inhibit gene expression. However, the functions of LkERF, an EAR motif-containing protein from Larix kaempferi, especially in reactive oxygen species (ROS) homeostasis, are not well understood. In the present research, we introduce a novel transcription factor, LkERF6, which contains an EAR motif and positively regulates gene expression, thereby enhancing drought and salt tolerance in tobacco. LkERF6 is classified within the ERF-B1 subfamily due to its conserved AP2/ERF domain and EAR motif. Subcellular localization assays demonstrated LkERF6 is primarily localized in the nucleus. Further analysis revealed that LkERF6 interacts with GCC and DRE elements and is significantly induced by NaCl and PEG6000. Moreover, LkERF6 transgenic tobacco plants exhibit lower ROS accumulation and higher levels of antioxidant enzyme activities. Additionally, correlation analysis identified a strong association between LkERF6 and three genes: LkSOD, LkCCS, and LkCAT. Y1H, EMAS, and DLR assays confirmed that LkERF6 directly interacts with the promoters of these genes through GCC-box and DRE-box to activate their expression. These findings shed new light on the function of EAR motif-containing transcription factors and highlight LkERF6’s crucial role in enhancing abiotic stress resistance by activating multiple ROS clearance genes.

Randomized trial of dietary acid reduction and acid-base status of patients with CKD and normal eGFR

IntroductionModern acid-producing diets in patients with stage G3-G5 chronic kidney disease (CKD) can cause severe acid accumulation with metabolic acidosis and less severe accumulation causing eubicarbonatemic acidosis in stages G2-G3, each with kidney injury. The impact of these diets on acid accumulation in those with CKD but normal eGFR (G1) is unclear. MethodsWe assessed whether acid accumulation occurs in patients with CKD and normal eGFR, and if added base-producing fruits and vegetables (F&V) or oral NaHCO3 (HCO3-) reduces acid accumulation and/or lowers kidney injury. We randomized 153 macroalbuminuric, non-diabetic, G1 (mean eGFR = 101 ml/min/1.73 m2) participants with hypertension-associated CKD to receive F&Vs in amounts to reduce dietary acid intake 50% (F&V, n=51), oral NaHCO3 to match alkali intake of F&V (HCO3-, n=51), or usual care (UC, n=51) for five years. We assessed acid accumulation by comparing observed to expected increase in plasma total CO2 in response to retained bicarbonate (dose minus urine bicarbonate excretion) two hours after an oral NaHCO3 bolus. ResultsBaseline acid accumulation, eGFR, urine excretion of albumin, N-acetyl-β-D-glucosamine, and angiotensinogen were not different among groups. Five-year acid accumulation [mean(SD)] was lower in F&V [-1.2 (11.0) mmoles] and HCO3- [-1.7 (10.8) mmoles] than UC [5.2 (10.3) mmoles, p<0.003)], consistent with lower acid accumulation in F&V and HCO3-. Five-year urine excretion of albumin, N-acetyl-β-D-glucosamine, and angiotensinogen were lower in F&V and HCO3- than UC, consistent with less kidney injury. ConclusionsDietary acid reduction reduces acid accumulation and kidney injury in patients with CKD and normal eGFR.

Safety, pharmacokinetics, and pharmacodynamics of sofnobrutinib, a novel non-covalent BTK inhibitor, in healthy subjects: First-in-human phase I study.

Bruton's tyrosine kinase (BTK) is a potential therapeutic target for allergic and autoimmune diseases. This first-in-human phase I study evaluated safety, pharmacokinetic, and pharmacodynamic profiles of sofnobrutinib (formerly AS-0871), a highly selective, orally available, non-covalent BTK inhibitor, in healthy adult subjects. Single ascending doses (SAD; 5-900 mg) and multiple ascending doses (MAD; 50-300 mg twice daily [b.i.d.] for 14 days [morning dose only on Day 14]) of sofnobrutinib were tested. In the entire study, all adverse events (AEs) were mild or moderate, and no apparent dose-proportional trend in severity or frequency was observed. No serious treatment-emergent AEs, cardiac arrythmias, or bleeding-related AEs were reported. In the SAD part, sofnobrutinib exhibited approximately dose-dependent systemic exposures up to 900 mg with rapid absorption (median time to maximum concentration of 2.50-4.00 h) and gradual decline (mean half-lives of 3.7-9.0 h). In the MAD part, sofnobrutinib showed low accumulation after multiple dosing (mean accumulation ratios of ≤1.54) and reached a steady state on ≤Day 7. Single dosing of sofnobrutinib rapidly and dose-dependently suppressed basophil and B-cell activations in exvivo whole blood assays. Multiple dosing of sofnobrutinib achieved 50.8%-79.4%, 67.6%-93.6%, and 90.1%-98.0% inhibition of basophil activation during the dosing interval of 50, 150, and 300 mg b.i.d., respectively. Based on pharmacokinetic-pharmacodynamic analysis, half-maximal inhibitory concentration (IC50) of sofnobrutinib for basophil activation was 54.06 and 57.01 ng/mL in the SAD and MAD parts, respectively. Similarly, IC50 for B-cell activation was 187.21 ng/mL. These data support further investigation of sofnobrutinib in allergic and autoimmune diseases.

Mutation of the K+ transporter SlHAK5 of tomato alters pistil morphology, ionome, metabolome and transcriptome in flowers.

K+ accumulation in plant tissues is a crucial factor for plant growth and development. The tomato high-affinity K+ transporter SlHAK5 is essential for root K+ acquisition from low external concentrations. It is also involved in K+ accumulation in pollen and plant fertility as slhak5 KO plants show a low rate of pollen germination, impaired pollen tube growth and parthenocarpic fruits. Here, we present a thorough analysis of slhak5 flowers, which showed relevant defects at the anatomic, ionomic, metabolomic and transcriptomic levels. First, slhak5 flowers exhibited shorter styles and enlarged ovaries that, together with a low number of seeds in fruits from slhak5 X WT crosses, indicated an effect of the slhak5 mutation on female fertility. Second, a lower accumulation of Ca2+, as well as of several metabolites such as amino acids, citric acid and sugars, was observed in mutant flowers, whereas indole-3-acetic acid content was increased when compared to the wild-type. Third, RNAseq conducted on pistils and stamens of wild-type and slhak5 plants revealed that transport and signalling pathways are significantly enriched in the gene expression analyses of stamens. Thus, it can be concluded that a functional SlHAK5 transporter is required to maintain appropriate Ca2+, metabolite and gene expression levels in flowers, and its absence leads to important reductions in both male and female fertility.

MsFtsH8 Enhances the Tolerance of PEG-Simulated Drought Stress by Boosting Antioxidant Capacity in Medicago sativa L.

Drought is a major abiotic stress that limits the growth and yield of alfalfa, a vital forage legume. The plant metalloproteinase Filamentation temperature-sensitive H (FtsH) is an ATP- and Zn2+-dependent enzyme that plays a significant character in the plant's response to environmental stress. However, its functional role in drought resistance remains largely unexplored. This study investigates the drought tolerance role of alfalfa MsFtsH8 by analyzing the growth, physiology, and gene expression of overexpressing plants under drought conditions. The results demonstrated that both MsFtsH8-overexpressing Arabidopsis and alfalfa plants exhibited superior growth condition and enhanced membrane stability. The overexpressing alfalfa plants also showed reduced MDA levels, higher proline content, lower H2O2 accumulation, an increased activity of antioxidant-related enzymes (SOD, POD, and CAT) activity, and an elevated expression of antioxidant-related genes. These results indicated that the overexpression of MsFtsH8 enhanced growth, improved osmotic regulation, reduced ROS levels, and increased antioxidative capacity, ultimately leading to greater drought tolerance in alfalfa. Our findings suggest that MsFtsH8 mitigates oxidative damage caused by drought by modulating the plant's antioxidant system, thus improving drought tolerance in alfalfa. This study provides a molecular basis and candidate genes for enhancing drought resistance in alfalfa through genetic engineering.

Supramolecular Self-Assembled Hydrogel for Antiviral Therapy through Glycyrrhizic Acid-Enhanced Zinc Absorption and Intracellular Accumulation.

Respiratory syncytial virus (RSV) is a common pathogen that causes respiratory infections in infants and children worldwide, significantly impacting hospitalization rates in this age group. Zinc ions are considered to have broad-spectrum antiviral potential against RNA viruses, including RSV. However, poor organism absorption and low intracellular accumulation of zinc require repeated high-dose supplementation, which may lead to unnecessary toxic side effects. In this research, a Zn2+-mediated glycyrrhizinic acid (GA)-based hydrogel (ZnGA Gel) was introduced and potentially developed to be a clinically available drug candidate for RSV therapy. ZnGA Gel was fabricated based on the cooperation of two potential RSV inhibiting molecules (Zn2+ and GA), where Zn2+ promoted self-assembly of GA and reduced its gel concentration and GA promoted zinc absorption and distribution in lung tissue in vivo. The facile construction of supramolecular hydrogel by the self-assembled coordination complex made it an injectable, temperature-sensitive, and pH-responsive controlled-release drug delivery for Zn2+. Most importantly, GA was observed to enhance organism absorption and intracellular accumulation of Zn2+ and was identified as a zinc ionophore for the first time. GA can colonize on the cell membrane and disturb cell membrane potential, resulting in an enhanced cell membrane permeability. In the presence of GA, more than 4.7-fold increasing Zn2+ concentrations materialized in the intracellular cytoplasm, compared to Zn2+ alone administration. This intracellular Zn2+ accumulation directly boosted the antiviral activities through improved inhibition of RSV replication-associated proteins and significantly inhibited RSV replication. Oral administration of ZnGA Gel on the RSV-infected mice model achieved an ideal therapeutic effect by effectively lowering viral load in the lungs, alleviating lung injury symptoms, and reducing inflammatory cell infiltration at pathological sites. The mechanism involved the inhibition of RSV replication-related proteins, aligning with our in vitro results. Additionally, ZnGA Gel had demonstrated biocompatibility, and reasonable supplementation of zinc was acceptable and effective for infants and children in clinical practice. Hence, the ZnGA Gel developed by us holds promise as an effective anti-RSV medicine in the future.

Silencing of SlMYB78-like Reduces the Tolerance to Drought and Salt Stress via the ABA Pathway in Tomato.

The MYB transcription factor family plays a crucial regulatory role in plant growth, development, biological progress, and stress responses. Here, we identified a R2R3-MYB transcription factor gene, SlMYB78-like, from tomato and characterized its function by gene silencing via RNA interference (RNAi). The results exhibited that the silencing of SlMYB78-like reduced the sensitivity of tomato seedlings to exogenous ABA. In addition, when exposed to drought and salt stresses, the RNAi lines grown in soil showed decreased tolerance, with lower ABA accumulation, relative water content, and chlorophyll content while displaying higher relative conductivity and malondialdehyde (MDA) content than the wild type. Moreover, the expression of genes related to chlorophyll biosynthesis, photosynthesis, and ABA biosynthesis/response were down-regulated in SlMYB78-like-silenced lines. Notably, the transcript level of SlCYP707-A2, which encodes a protein involved in ABA degradation, was up-regulated significantly after stresses. The transient expression assay Dual-luciferase (Dual-LUC) and a yeast one-hybrid (Y1H) assay demonstrated that SlMYB78-like bound to the promoter of SlCYP707-A2. Additionally, the physical interaction between SlMYB78-like and SlDREB3, which functioned in ABA signaling transduction, was identified through yeast two-hybrid (Y2H) and bimolecular fluorescence complementation (BiFC) assays. Collectively, our study illustrates that SlMYB78-like participates in the abiotic stress response via the ABA pathway.

Head-to-head comparison of 68Ga-FAPI-46 PET/CT, 18F-FDG PET/CT, and contrast-enhanced CT for the detection of various tumors.

FAPI-PET/CT exhibits high tumor uptake and low background accumulation, enabling high-sensitivity tumor detection. We compared the diagnostic performance of 68Ga-FAPI-46 PET/CT plus contrast-enhanced CT (CE-CT), 18F-FDG PET/CT plus CE-CT, and standalone CE-CT in patients with various malignancies. 232 patients underwent 68Ga-FAPI-46 PET/CT,18F-FDG PET/CT, and CE-CT each within 4weeks. Detection rates were assessed by a blinded reader, with ≥ 2weeks between scans of the same patient to avoid recall bias. A sub-analysis of diagnostic performance was performed for 490 histopathologically validated lesions. Detection rates were compared using McNemar's test. Lesion-based detection rates in 68Ga-FAPI-46 PET/CT plus CE-CT, 18F-FDG PET/CT plus CE-CT, and CE-CT alone were 91.2% (1540/1688), 82.5% (1393/1688) and 60.2% (1016/1688). The detection rates were significantly higher for 68Ga-FAPI-46 PET/CT plus CE-CT than for 18F-FDG PET/CT plus CE-CT (p < 0.02 for primary lesions and p < 0.001 for total, abdominopelvic nodal, liver and other visceral lesions) and CE-CT (p < 0.0001 for total, primary, cervicothoracic nodal, abdominopelvic nodal, liver, other visceral, and bone lesions). In the sub-analysis, sensitivity, specificity, positive and negative predictive value, and accuracy were 61.3%, 96.7%, 81.4%, 91.4% and 90.0% for 68Ga-FAPI-46 PET/CT plus CE-CT, 57.0%, 95.7%, 75.7%, 90.5% and 88.4% for 18F-FDG PET/CT plus CE-CT, and 51.6%, 97.2%, 81.4%, 89.6% and 88.6% for CECT, respectively. 68Ga-FAPI-46 PET/CT plus CE-CT demonstrates a higher tumor detection rate than 18F-FDG PET/CT plus CE-CT and CE-CT in a diverse spectrum of malignancies, especially for primary, abdominopelvic nodal, liver, and other visceral lesions. Further studies on which entities draw particular benefit from 68Ga-FAPI-46 PET/CT are warranted to aid appropriate diagnostic workup. A total of N = 232 patients were analyzed. Of these, N = 50 patients were included in a prospective interventional trial (NCT05160051), and N = 175 in a prospective observational trial (NCT04571086) for correlation and clinical follow-up of PET findings; N = 7 patients were analyzed retrospectively.

Using Bioindicators to Assess Heavy Metal Contamination in Surface Waters and Public Health Impacts

Objective: This study aims to explore the use of bioindicators for assessing metal contamination in surface waters and its effects on public health. Theoretical Framework: The persistence and variation of heavy metal contamination in the Ave River Basin over 15 years underscore the need for ongoing monitoring and intervention. The detection of elevated levels of chromium, cadmium, lead, and zinc in aquatic mosses highlights the continuing impact of industrial activities, particularly metal coating facilities, on the river's ecosystem. Method: In the study area, the Ave River and some of its tributaries, a monitoring programme of twelve sampling points was defined in two campaigns and eight metallic ions were determined in the laboratory by atomic absorption spectrophotometry (Cd, Pb, Cu, Cr, Fe, Hg, Ni and Zn). Results and Discussion: The highest metal accumulation in mosses was observed for iron (Fe), whereas the lowest accumulation was found for mercury (Hg) during Campaign II. The order of metal accumulation in the moss samples, from highest to lowest, was Fe &gt; Zn &gt; Cu &gt; Cr &gt; Ni &gt; Pb &gt; Cd &gt; Hg. The Metal Pollution Index (MPI) revealed changes in contamination levels between campaigns. Research Implications: The contamination patterns suggest the influence of industrial activities, particularly metal coating facilities. Monitoring and mitigation efforts are necessary to address persistent heavy metal pollution in the Ave River Basin. Originality/Value: The results of this research will contribute to a better understanding of the sources and loads of pollutants discharged and responsible for the contaminants. This can be used to discourage potential polluters and better manage the basin's water resources and possible risks to human health.

Grain Lysine enrichment and improved stress tolerance in rice through protein engineering.

Amino acids are a major source of nourishment for people living in regions where rice is a staple food. However, rice grain is deficient in essential amino acids, such as lysine. The activity of dihydrodipicolinate synthase (DHDPS) enzyme is crucial for lysine production in higher plants, but it is highly regulated through a feedback inhibition by its end product lysine, leading to its limited activity in the grain and resulting in low lysine accumulation. We identified lysine binding sites in the DHDPS enzyme and introduced key mutations to make it lysine feedback insensitive. Using in vivo analysis and functional complementation assays, we confirmed that protein engineering of the DHDPS renders it insensitive to lysine. Expression of mutated DHDPS resulted in 29 % higher lysine and 15 % higher protein accumulation in rice grains than the wild type. Importantly, the lysine content in transgenic grains was maintained in cooked rice. Further, the transgenic plants exhibited enhanced stress tolerance along with better antioxidant levels, improved photosynthesis, and higher grain yield compared to wild type plants. We have shown for the first time in rice that protein engineering of DHDPS can lead to accumulation of lysine in grains and impart abiotic stress tolerance. This approach could improve health in regions with nutrient deficiencies and environmental stressors that challenge food production and human health.

Spent brewer yeast glucan improves metabolic parameters and inflammatory markers reducing health risky changes imposed by the consumption of hypercaloric diet

β-glucans from yeasts are prebiotic health-promoting food ingredients able to boost immune response. An environmentally friendly source of yeast glucans is the spent brewer's yeast. This study investigated the effects of the administration of carboxymethyl glucan (CMG) (20 mg/kg, 12 weeks, orally), a derivative of the β-glucan from spent brewer's yeast, on food intake, weight gain, dyslipidemia, glucose intolerance, pro-inflammatory state, and hepatic steatosis of rats receiving a hypercaloric diet. The animals were divided into three groups: Normocaloric control (NCT); Hypercaloric diet and saline (HCT); and Hypercaloric diet and CMG (H-CMG). CMG administration could reduce rats' caloric value and food intake, reducing the index of adiposity (HCT:7.2±0.4% vs H-CMG:5.3±0.4% p < 0.05) and weight gain (HCT:111.1±3.8g vs H-CMG: 68.3±4.9g p < 0.05). Furthermore, it improved lipemic and glycemic profiles and lowered hepatic lipid accumulation. CMG decreased Monocyte Chemoattractant Protein (MCP)-1 (H-CT: 89.5±1.8 pg/mL vs H-CMG: 50.7±12.2 pg/mL p < 0.05), Interleukin (IL)-12 (H-CT: 206.0±14.0 pg/mL vs H-CMG:132.4±19.9 pg/mL, p < 0.05), and C-reactive protein (CRP) levels and increased IL-10 levels, decreasing the cardiovascular risk and pro-inflammatory state. Therefore, CMG has beneficial effects in preventing weight gain and, consequently, improves the metabolic profile in rats while consuming a hypercaloric diet. Findings indicate the spent brewer's yeast CMG can be a strategy to attenuate the risky metabolic changes related to the consumption of a hypercaloric diet.