Search Results (3,354)

The formulation of rhizobial bioinoculants remains a critical bottleneck for the large-scale deployment of biological nitrogen fixation in sustainable agriculture, mainly due to limitations in the stability and viability of conventional liquid products. In this study, a spray-drying-based process was developed and optimized to produce a stable and functional bioinoculant using Ensifer meliloti Rm8530, an EPS II–producing strain with enhanced stress tolerance. Strain robustness was evaluated through thermal and osmotic stress assays, together with growth performance across relevant temperature and pH ranges. Six carrier-based formulations combining polysaccharides and proteins were then tested under controlled spray-drying conditions. Process performance was assessed in terms of powder recovery, residual moisture, bacterial survival, yield, and storage stability over 16 weeks. The morphological integrity of spray-dried particles and rehydrated cells was analyzed by scanning electron microscopy. The biological functionality of selected formulations was subsequently validated in planta using alfalfa as a host model. Among the formulations tested, a mixed alginate–gum Arabic matrix showed the best overall balance between process efficiency, post-drying viability, long-term stability, and symbiotic performance. Spray-dried cells retained the ability to induce nodulation and support early plant responses under the conditions evaluated. These results demonstrate that spray drying, combined with appropriate strain selection and formulation design, constitutes a viable and scalable platform for producing stable, functional rhizobial bioinoculants. Full article

Background: Caffeic acid (CA), with antioxidant and immunomodulatory properties, was formulated in liposomes to increase its efficacy. The study targets triple-negative breast cancer (TNBC), characterized by the absence of ER, PR, and HER2 receptors. Methods: For CA-loaded liposomes, the pharmacological effects on TNBC cell lines, parental Hs578T (HS) and Doxorubicin-resistant Hs578T (HSD) cells were evaluated by determining the cell growth inhibition ratio measured by the (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay, oxidative stress, apoptosis rate, membrane damage and transcription factor expressions, and DNA damage, with or without exposure to Doxorubicin (Dox). The Results: demonstrated that CA-loaded liposomes were stable and had high entrapment capacity. They exerted apoptotic effects on both cells, comparable to Doxorubicin, and increased cell membrane damage. The liposomes increased STAT3 expression in HS cells, while they reduced NRF2 and STAT3 in HSD cells, suggesting beneficial effects on Dox-resistant breast tumor cells. In HS cells exposed to Dox, CA treatment improved the number of viable tumor cells and decreased the rate of apoptosis, while in HSD cells it enhanced apoptosis as a mechanism of cell death and decreased pro-survival molecules, STAT3 expression in parallel with reduced NRF2 activation. Conclusions: The results indicated that CA encapsulated in liposomes was able to interfere with some survival mechanisms of triple-negative cells and could inhibit their proliferation. Full article

Sporulation represents a complex metabolic reprogramming process in bacteria. In this study, we used CRISPR-Cpf1 to knock out spoIIE and rsfA in Bacillus licheniformis. The ΔspoIIE strain completely lost sporulataion capacity, while ΔrsfA showed a 25% reduction. Although viable cell counts decreased by 80.7% and 45.7%, respectively, glucose consumption and 2,3-butanediol synthesis remained unchanged, and acetoin synthesis increased by 19% in ΔspoIIE. Per-cell metabolic rates were significantly enhanced: glucose uptake increased 2.7–3.4-fold, acetoin synthesis 2.3–4.2-fold, 2,3-butanediol synthesis 1.7-fold, and heterologous protein expression 10–15-fold. These findings demonstrate that blocking sporulation liberates metabolic resources and enhances the specific productivity of vegetative cells, providing a strategy for engineering high-performance B. licheniformis cell factories. Full article

Programmed cell death (PCD) is a pivotal biological process that occurs at various stages of plant development, including embryogenesis and seed germination. This study investigated whether the absence of PCD in endosperm cells is connected to the poor germination of Viola odorata seeds. Seeds of poorly germinating V. odorata and well-germinating V. × wittrockiana were either cold-stratified for 10 days or left untreated. Germination tests, tetrazolium viability tests, Western blot analyses for caspase-like proteases, and Terminal deoxynucleotidyl transferase (TdT) dUTP nick end labeling (TUNEL) assays for DNA strand break detection were performed. The results revealed that V. odorata seeds did not germinate, regardless of stratification or lack thereof, whereas in V. × wittrockiana, stratification significantly increased their germination capacity (34 ± 6.5% vs. 56.5 ± 9.8% in non-stratified and stratified seeds, respectively). The tetrazolium viability test revealed that V. odorata seeds were nonviable (100% nonviable endosperm and 96% nonviable embryos in total), whereas the seeds of V. × wittrockiana were highly viable (63% and 59% endosperm and embryos in total, respectively). Western blot analysis revealed that in the germinating seeds of V. × wittrockiana, caspase-like activity was detected in the endosperm but not in the embryos, whereas in seeds that failed to germinate, the PCD signal in the endosperm was very weak. In the seeds of V. odorata, caspase-like activity was detected in the embryos and endosperm collected directly after 10 days of stratification, but no signal was detected in the seeds left to germinate for one month after cold stratification. TUNEL assays revealed DNA strand breaks in the peripheral part of the endosperm in V. odorata and in non-germinating V. × wittrockiana, whereas in the germinating seeds of V. × wittrockiana, DNA strand breaks were detected in the endosperm cells adjacent to the embryo. These findings indicate that endosperm-localized PCD facilitates nutrient mobilization to the embryo and seems crucial for successful germination. Overall, these results suggest that PCD contributes to the regulation of seed germination in Viola spp. Full article

Background/Objectives: Chronic traumatic spinal cord injury (SCI) causes persistent neurological deficits for which no clinically effective regenerative therapy is currently available. Mesenchymal stromal/stem cells (MSCs), particularly Wharton’s jelly-derived MSCs (WJ-MSCs), demonstrate immunomodulatory and neurotrophic potential. This phase I/II study evaluated the safety and efficacy of intrathecal allogeneic WJ-MSC administration in individuals with chronic incomplete cervical SCI. Methods: In this multicentre, randomised, double-blind, placebo-controlled trial (NCT05054803, EudraCT 2021-000346-18), 18 participants with chronic (1–5 years post-injury) incomplete cervical SCI (AIS B–D) received two intrathecal injections of WJ-MSCs (0.7–1.3 × 10⁶ viable cells/kg) or a placebo at baseline and 3 months. Seventeen participants completed the 12-month follow-up. Primary outcomes assessed safety, and secondary endpoints included International Standards for Neurological Classification of Spinal Cord Injury (ISNCSCI) motor and sensory scores, spasticity, neuropathic pain, functional independence, neurophysiological measures, and quality of life. Results: Intrathecal WJ-MSC administration was safe and well tolerated. Eighty adverse events occurred (placebo: 26; WJ-MSC: 54), predominantly mild or moderate; four severe events were unrelated to treatment. Both groups demonstrated significant within-group improvements in total motor scores at 12 months, with no between-group difference. No treatment effects were observed for sensory scores, electrophysiological measures, functional independence, spasticity, pain, or patient-reported outcomes. Conclusions: In this first randomised, placebo-controlled trial evaluating intrathecal WJ-MSCs in chronic incomplete cervical SCI, WJ-MSC administration demonstrated a favourable safety profile; however, no significant between-group differences were detected relative to the placebo. Given the limited sample size and early-phase design, the efficacy findings should be interpreted cautiously. Future research should explore enhanced cell products, intensified dosing schedules, optimised delivery strategies, early intervention, and multimodal therapeutic combinations. Full article

Van Maldergem Syndrome (VMS) is a rare autosomal recessive disorder caused by pathogenic variants in the atypical cadherin genes DCHS1 or FAT4 and is marked by craniofacial, skeletal, and neurodevelopmental abnormalities. Although DCHS1–FAT4 binding is mediated by their respective extracellular domains, the in vivo function of the DCHS1 intracellular domain (ICD) is poorly defined. To test its function, we generated mice in which the DCHS1 ICD was deleted and replaced with a V5 epitope tag (Dchs1^ΔICD-V5). Homozygous Dchs1^{ΔICD-V5/ΔICD-V5} mice are viable but exhibit VMS-like craniofacial flattening with enlarged fontanelles and reduced palatine/maxillary structures, along with airway cartilage abnormalities including reduced mineralization and decreased tracheal circularity. In periventricular regions, wild-type DCHS1 expression shows polarized localization, whereas mice with the ICD deletion exhibit altered cell polarization within the subventricular zone, concomitant with changes in neural cellular distribution. Neonatal brains display reduced pYAP1: YAP1 ratios and increased Ki67+ proliferation with greater Ki67–neuronal co-localization within the periventricular zone. Together, these data identify the DCHS1 ICD as a critical effector for DCHS1 signaling and a regulator of polarity-dependent growth, with associated changes in Hippo pathway activity during craniofacial and neural morphogenesis. Additionally, our data establish Dchs1^{ΔICD-V5/ΔICD-V5} mice as a model that recapitulates core features of VMS, thereby allowing new mechanistic discoveries into its pathogenesis. Full article

The large-scale production of microbial bioplastics remains limited by high production costs, reliance on refined substrates, and inefficient utilization of agro-industrial residues. Although sugarcane bagasse has been explored as a carbon source for polyhydroxyalkanoate production, studies have predominantly focused on poly (3-hydroxybutyrate) (PHB), with limited reports on copolymer synthesis from pentose-rich lignocellulosic streams. In this study, a newly isolated Bacillus sp. HLI02 was employed for the biosynthesis of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), using pentosan-rich sugarcane bagasse hydrolysate as an inexpensive and sustainable carbon source. Fermentation parameters were systematically optimized at different pH and temperature, and the strain demonstrated efficient conversion of xylose-rich hydrolysate into PHBV without the requirement for external nutrient supplementation. Under optimized conditions (pH 7.0, 37 °C, and C/N ratio of 40), a maximum PHBV yield of 2 g/L, corresponding to 59.5% of cell dry weight, was achieved. Structural and compositional analyses using Fourier transform infrared spectroscopy (FTIR), gel permeation chromatography (GPC), and ¹H and ¹³C nuclear magnetic resonance (NMR) spectroscopy confirmed successful PHBV copolymer formation with well-defined structural characteristics. Thermal analysis revealed a decomposition temperature of 166 °C, indicating good thermal stability. The produced PHBV further exhibited favourable biocompatibility and biodegradability, supporting its potential applicability in sustainable packaging and related sectors. This work demonstrates the effective conversion of hemicellulosic sugarcane bagasse hydrolysate into PHBV using a newly isolated Bacillus strain, highlighting an underexplored route for copolymer production from agro-waste–derived C5 sugars. By integrating low-cost feedstock utilization with process optimization and comprehensive polymer characterization, this study contributes to the development of economically viable and sustainable bio-based polymer production strategies. Full article

Monkeypox virus, a zoonotic DNA virus belonging to the Orthopoxvirus genus, has emerged as a global health issue because of its fast spread to 104 nations over six continents. In the current study, an immunoinformatics pipeline was used to design a multiepitope-based prophylactic vaccine targeting the A35R glycoprotein and E8L membrane proteins of the monkeypox virus. Selected target proteins were surface-exposed, non-homologous to the human proteome, and essential for viral pathogenesis. B-cell and T-cell (MHC-I and MHC-II) epitopes with high antigenicity (>0.5), non-allergenicity, non-toxicity, and highly soluble in water with strong affinity towards innate and adaptive receptors, were prioritized. Shortlisted epitopes were combined to design the final vaccine utilizing an adjuvant (50S ribosomal L7/L12) and appropriate linkers for improved immunogenicity. Population coverage analysis showed wide HLA representation with 83.57% (MHC-I) and 88.8% (MHC-II) global coverage, including 89.6% for West Africa and 87.3% for Central Africa. Docking analysis of the vaccine construct with the TLR-4 receptor revealed stable interactions (−695.6 kcal/mol). Molecular dynamics simulations and binding free energies further confirmed structural stability. Immune simulations predicted strong activation of both humoral and cellular immune responses. These results indicate that the designed multiepitope vaccine construct is a viable option for additional experimental validation against the monkeypox virus. Full article

The transportation sector’s decarbonization represents one of the most critical challenges in achieving global climate targets. This study presents a comprehensive analysis of an integrated renewable energy system that produces green hydrogen through a hybrid solar photovoltaic (PV) and wind power configuration. The proposed system combines a 1.2 MWp solar array with 800 kW wind turbines, feeding a 1 MW proton exchange membrane (PEM) electrolyzer for hydrogen production. The hydrogen is subsequently compressed, stored at 350 (for trucks and buses) and 700 bar (for cars), and then utilized either directly for fuel cell electric vehicles (FCEVs) or reconverted to electricity via a 250 kW stationary PEM fuel cell to support electric vehicle (EV) charging infrastructure. Through detailed techno-economic simulation using HOMER Pro and MATLAB/Simulink 2022a, we demonstrate that the hybrid configuration achieves a 71% electrolyzer capacity factor, producing 55.8 tonnes of hydrogen annually with a levelized cost of 5.82 €/kg. The system ensures over 60 h of grid-independent operation while reducing CO₂ emissions by 1656 tones annually compared to conventional grid-powered alternatives. Results indicate that hybrid renewable hydrogen systems can provide economically viable solutions for sustainable mobility infrastructure, with projected cost reductions making them competitive with fossil fuel alternatives by 2030. Full article

Background/Objectives: Active surveillance (AS) is a viable option for patients with low-risk/low-burden prostate cancer (PCa). Approximately 40–50% of patients will develop disease progression and conversion to active treatment. Therefore, better risk stratification may aid patients and urologists to improve decision making. Herein, the proliferation marker Ki-67 was examined for its prognostic potential in AS patients. Methods: Fifty-nine patients were included. Median follow-up time was 58 months (range, 10–162 months). Tumor-bearing biopsies were evaluated using immunohistochemistry (IHC) staining for Ki-67 and evaluated using digital imaging analysis to determine the percentage of Ki-67-positive PCa cells per biopsy. Results: Thirty-three of 59 patients (55.9%) developed progression. Thirty-one of 59 patients (52.5%) showed Ki-67-positive biopsies (median 0.8%; range, 0–11.9%). The median of Ki-67-positive cells was 1.5% (range, 0–11.9%) in patients with and 0% (range, 0–6.3%) in patients without progression. Comparing patients with Ki-67-positive and Ki-67-negative biopsies showed a worse progression free survival (PFS) in patients with Ki-67-positive biopsies after a period of 15 months, however, without reaching statistical significance (p = 0.071). A 5% threshold for Ki-67 positivity led to a significant difference in PFS. Further exploratory analysis revealed that patients with Ki-67-positive biopsies and aged ≥65 years or with >1 tumor-bearing biopsy show a significantly worse outcome (p = 0.038 and p = 0.037, respectively). Conclusions: Our results suggest that patients with Ki-67-positive biopsies remaining in AS for >1 year have an increased risk for PCa progression and conversion to treatment. Studies to further confirm Ki-67 as a marker for risk stratification, especially with a positivity cut-off of 5%, are warranted in larger cohorts of AS patients. Full article

Objectives: The identification of novel antimicrobial agents for use in root canal treatment may provide opportunities to improve treatment outcomes. This study aimed to assess the antimicrobial efficacy of different molecular weights of chitosan (CS), and how modification with CS may impact on the antimicrobial, physico-mechanical and biological properties of Biodentine™, a calcium-silicate-based material used in endodontics. Methods: C. albicans biofilms were treated with either 3% sodium hypochlorite (NaOCl) or a 0.05% or 0.1% CS solution for 5 min. The growth medium was replenished, and cells were re-incubated for additional 72 h. Regrowth of biofilms was assessed using a colorimetric XTT assay. Additionally, multispecies biofilms were established and the regrowth of biofilms on Biodentine discs were quantified following the addition of 0.5 wt% and 1 wt% of CS powder using qPCR. The physico-mechanical and biological properties of the new composite of Biodentine and CS were also evaluated. Results: Viability readings revealed significant initial biofilm inhibitory effects of CS solutions, followed by significant regrowth after 72 h. Upon the addition of CS to Biodentine, significant reductions in multispecies biofilm regrowth were determined. Notably, the antibiofilm activity of CS was found to be increased as the molecular weight decreased. The addition of powdered CS of low molecular weight showed a reduction in the mechanical properties of Biodentine, whereas no detrimental effects on the other material properties were noted. Conclusions: Chitosan may not be useful as an alternative irrigant to NaOCl. Addition of CS to Biodentine represents a potential means of augmenting the antimicrobial activity of Biodentine against persistent microorganisms following endodontic therapy. Despite the reductions in mechanical properties of the material, the new composite still represents a viable material option when material strength and hardness are not critical. Full article

Lactiplantibacillus plantarum EL2, isolated from traditional fermented yak milk in the high-altitude Gannan Tibetan Autonomous Prefecture, produces the class IIb bacteriocin PlnJK. This study established three distinct cultivation models that critically influenced bacteriocin yield. Microbial co-culture was found to enhance the stress tolerance of EL2, significantly boosting PlnJK production. The optimal inducing strain, Enterococcus faecalis MH2, increased the bacteriocin inhibition zone diameter from 15.38 mm to 25.58 mm. Following optimization of key parameters—initial inoculum concentration (10⁷ CFU/mL), inoculation ratio (3:1, EL2:MH2), and initial pH (6.0)—the inhibition zone diameter reached 30.32 mm, representing a 1.97-fold increase over pure culture. Co-culture not only advanced the onset but also extended the duration of bacteriocin synthesis. Throughout the 24 h incubation, cell density, AI-2 autoinducer concentration, and the expression of key regulatory genes were significantly elevated in co-culture compared to monoculture, aligning with a cell-density-dependent, quorum-sensing (QS) regulatory paradigm. Bacteriocin production was co-regulated by two QS pathways: the AI-2/luxS system and the plnA-mediated autoinducing peptide (AIP). Gene expression analysis revealed differential temporal regulation: luxS expression was higher during the exponential phase (2.29 vs. 1.42 in stationary phase), while plnA exhibited the opposite pattern (1.42 in exponential vs. 2.21 in stationary phase). This indicates that the AI-2/luxS pathway drives strong induction during active growth, whereas plnA/AIP-mediated promotion becomes predominant later. The stationary-phase effect is likely triggered by the accumulation of specific MH2 metabolites, which impose an environmental stress on EL2, stimulating the pln-encoded regulatory system and further enhancing bacteriocin yield. This work provides an economically viable strategy and a novel theoretical framework for optimizing microbial cultivation, enhancing bacteriocin production, and elucidating the complex QS-mediated regulatory mechanisms involved. Full article

Calcium-ion batteries (CIBs) offer several advantages. CIBs are viable alternatives to lithium-based battery systems owing to the natural abundance, low cost, and high volumetric capacity of calcium. However, their development has been severely constrained by electrolyte instability and water sensitivity. We conducted a systematic examination of Ca(ClO₄)₂ and Ca(PF₆)₂ electrolytes, focusing on low-cost salt production, solvent selection, and stringent dehydration procedures. Acetonitrile (ACN) was the ideal solvent for high salt solubility and reversible Ca²⁺ electrochemistry, while carbonate solvents failed rapidly. We found that even a small amount of moisture in the electrolyte significantly affected the electrochemical performance. This study improved the dehydration process by using 3 Å molecular sieve (MS3A) and vacuum drying to reduce moisture to ppm levels, stabilizing the electrolyte. Prussian blue (PB) half cells exhibited reversible capacities of up to ≈95 mAh g⁻¹, whereas PB-hard carbon full cells utilizing dried Ca(ClO₄)₂ showed stable cycling over 240 cycles with a Coulombic efficiency of ≈99% and capacity loss of only ≈17%. This study establishes a moisture-controlled electrolyte as a critical enabler for practical CIBs. Full article

Traumatic injury, stenosis, and malignancy involving large segments of the airway are difficult to reconstruct and require novel solutions. Despite advances in surgical techniques, the reconstruction of long-segment tracheal defects remains a significant challenge. Several bioengineering approaches have been explored for tracheal regeneration in vitro and in vivo, using cells in combination with three dimentional (3D) biological or synthetic scaffolds. This paper reviews recent advances in developing bioengineered trachea and the technologies utilized toward generating transplantable tracheal grafts. Specifically, the review will focus on the recellularization of tissue-engineered grafts using natural or synthetic scaffolds, highlighting relevant cell types used to reconstitute tracheal epithelium and cartilage. The promise of newly explored paradigms, including the application of pluripotent stem cells, will be discussed with an overview of associated challenges and necessary steps for future translation. Overall, these advances provide a foundation for the development of clinically viable tracheal grafts, bringing engineered tracheal reconstruction closer to reality. Full article

Neoadjuvant strategies in head and neck squamous cell carcinoma (HNSCC) are reshaping therapeutic paradigms by shifting emphasis from anatomical staging toward biology-driven response stratification. The transition from induction chemotherapy to immune checkpoint–based and combination regimens has transformed the perioperative setting into a translational platform that enables interrogation of tumor–immune interactions and clonal selection under therapeutic pressure prior to surgery. In this context, pathological response assessment has emerged as a robust surrogate endpoint, overcoming the limitations of radiologic evaluation, which often fails to capture immune-mediated pseudoprogression and spatially heterogeneous regression. Quantification of residual viable tumor (RVT) provides a reproducible metric of therapeutic efficacy, while characterization of immune-related regression beds, tertiary lymphoid structures, macrophage polarization states, and compartment-specific nodal responses offers mechanistic insight into tumor clearance and resistance evolution. Evidence from phase II trials, single-cell sequencing, spatial transcriptomics, and multiplex immune profiling supports the prognostic relevance of pathology-driven endpoints. Integration of digital pathology and artificial intelligence–assisted image analysis further enhances reproducibility and enables high-resolution mapping of residual disease and immune architecture. Within this modern oncologic framework, the neoadjuvant-treated specimen functions as a dynamic biomarker platform guiding response-adapted surgical strategies and biomarker-driven clinical trial design. This study was designed as a narrative review. A structured literature search was performed using PubMed and major oncology journals to identify relevant studies on pathology-driven response assessment in neoadjuvant-treated head and neck squamous cell carcinoma. The review focused on publications addressing histopathological response criteria, immune microenvironment remodeling, spatial profiling technologies, and computational pathology approaches. Full article