Metabolic Activity Research Articles

Tailoring metabolic activity assays for tumour-engineered 3D models

Monitoring cell behaviour in hydrogel-based 3D models is critical for assessing their growth and response to cytotoxic treatment. Resazurin-based PrestoBlue and AlamarBlue reagents are frequently used metabolic activity assays when determining cell responses. However, both assays are largely applied to cell monolayer cultures but yet to have a defined protocol for use in hydrogel-based 3D models. The assays' performance depends on the cell type, culture condition and measurement sensitivity. To better understand how both assays perform, we grew pancreatic cancer cells in gelatin methacryloyl and collagen hydrogels and evaluated their metabolic activity using different concentrations and incubation times of the PrestoBlue and AlamarBlue reagents. We tested reagent concentrations of 4 % and 10 % and incubation times of 45 min, 2 h and 4 h. In addition, we co-cultured cancer cells together with cancer-associated fibroblasts and peripheral blood mononuclear cells in gelatin methacryloyl hydrogels and subjected them to gemcitabine and nab-paclitaxel to evaluate how both assays perform when characterising cell responses upon drug treatment. CyQuant assays were conducted on the same samples and compared to data from the metabolic activity assays. In cancer monocultures, higher reagent concentration and incubation time increased fluorescent intensity. We found a reagent concentration of 10 % and an incubation time of 2 h suitable for all cell lines and both hydrogels. In multicellular 3D cultures, PrestoBlue and AlamarBlue assays detected similar cell responses upon drug treatment but overestimated cell growth. We recommend to assess cell viability and growth in conjunction with CyQuant assays that directly measure cell functions.

Electrospun Polylactic Acid/Polyethylene Glycol/ Silicate‐Chlorinated Bioactive Glass Composite Scaffolds for Potential Bone Regeneration

ABSTRACTThe integration of bioglass with polymers in tissue engineering scaffolds holds promise for enhancing bone regeneration. This study explores the fabrication and characterization of composite scaffolds comprising polylactic acid (PLA)/polyethylene glycol (PEG) fibers incorporated with silicate‐chlorinated bioglasses (45S5 and 58S). Electrospinning was utilized to produce the scaffolds, followed by physical–chemical and in vitro evaluations. Scanning electron microscopy (SEM) revealed uniform fiber formation, with bioglass incorporation observed in the composite groups. Bioglass incorporation led to a significant reduction in fiber diameter. Thermogravimetric analysis (TGA) estimated bioglass content, with 58S exhibiting the highest incorporation. Contact angle measurements indicated enhanced hydrophilicity in bioglass‐containing groups. In vitro, bioactivity assessment in simulated body fluid (SBF) demonstrated apatite formation potential and the pH variance indicates a slightly alkaline to neutral condition. Cell culture studies revealed robust cellular adhesion and metabolic activity across all groups, with no cytotoxic effects observed. Overall, these findings suggest the potential of PLA/PEG bioglass composite scaffolds for bone tissue engineering applications.

Insulin oxidation and oxidative modifications alter glucose uptake, cell metabolism, and inflammatory secretion profiles

Insulin participates in glucose homeostasis in the body and regulates glucose, protein, and lipid metabolism. Chronic hyperglycemia triggers oxidative stress and the generation of reactive oxygen species (ROS), leading to oxidized insulin variants. Oxidative protein modifications can cause functional changes or altered immunogenicity as known from the context of autoimmune disorders. However, studies on the biological function of native and oxidized insulin on glucose homeostasis and cellular function are lacking. Native insulin showed heterogenous effects on metabolic activity, proliferation, glucose carrier transporter (GLUT) 4, and insulin receptor (INSR) expression, as well as glucose uptake in cell lines of five different human tissues. Diverse ROS compositions produced by different gas plasma approaches enabled the investigations of variously modified insulin (oxIns) with individual oxidative post-translational modification (oxPTM) patterns as identified using high-resolution mass spectrometric analysis. Specific oxIns variants promoted cellular metabolism and proliferation in several cell lines investigated, and nitrogen plasma emission lines could be linked to insulin nitration and elevated glucose uptake. In addition, insulin oxidation modified blood glucose levels in the chicken embryos (in ovo), underlining the importance of assessing protein oxidation and function in health and disease.

Soil application potential of post-sorbents produced by co-sorption of humic substances and nutrients from sludge anaerobic digestion reject water

This study introduces a novel soil conditioning approach using humic substances (HSs) and nutrients co-recovered from reject water from sewage sludge anaerobic digestion. For the first time, HSs and nutrients were simultaneously recovered through sorption on low-cost, environmentally inert materials: natural rock opoka (OP) and waste autoclaved aerated concrete (WAAC). This innovative application of OP and WAAC as carriers and delivery agents for soil-relevant substances offers potential for resource recovery and soil conditioning. Results indicate that the post-sorption opoka (PS-OP) and post-sorption waste autoclaved aerated concrete (PS-WAAC) effectively release retained HSs at 350–480 μg g⁻1 d⁻1, respectively. These materials also show potential as NPK fertilizers, releasing 280–430 μg g⁻1 d⁻1 N-NH₄⁺, 80–150 μg g⁻1 d⁻1 P-PO₄³⁻, and 270–350 μg g⁻1 d⁻1 K⁺. Additionally, PS-OP demonstrated promising fungicide properties, reducing P. diachenii growth by 31% at a concentration of 1 g L⁻1. A two-way ANOVA indicated that the effects of PS-OP and PS-WAAC on soil physicochemical and biological parameters varied with plant species. Both post-sorbents improved the quality of soil collected from sand mining area, increasing cation exchange capacity by 7%–85% and organic matter content by 10%–58%. They also enhanced the functional potential of soil microbial communities, increasing their metabolic activities by 23%–36% in soils sown with clover and by 33%–39% in soils sown with rapeseed. An opposite effect was observed in soils sown with sorghum, suggesting these amendments may not universally act as plant biostimulants. The effectiveness of these post-sorbents in enhancing plant growth varied depending on plant species and the mineral base of the post-sorbent. PS-OP increased the total length of clover and sorghum by 41% and 36%, and their fresh biomass by 82% and 80%, respectively. In turn, PS-WAAC increased the total length of clover and sorghum by 76% and 17%, and their fresh biomass by 29% and 15%, respectively. It was notably more effective than PS-OP for rapeseed. This study proposes a strategy to decrease reliance on non-renewable resources and costly sorbents while minimizing environmental impact. It shows that PS-OP and PS-WAAC can enhance soil quality, microbial activity, and plant growth. Given their origins, these amendments are recommended for soil remediation, particularly in degraded areas. Future research should focus on optimizing their application across various plant species to maximize effectiveness.

Nanostructured silver vanadate gel: Evaluation of physicochemical, mechanical, and antibiofilm properties against a five-species oral model

The aim of this study was to develop a gel with nanostructured silver vanadate decorated with silver nanoparticles (β-AgVO3) and to evaluate its physicochemical, mechanical, and antibiofilm properties against a five-species oral model composed of Candida albicans, Staphylococcus aureus, Escherichia coli, Enterococcus faecalis, and Pseudomonas aeruginosa. The formulations were prepared with 0 % w/v (G0), 0.02 % w/v (G1), 0.05 % w/v (G2), and 0.12 % w/v (G3) β-AgVO3. The physicochemical properties of pH (n = 5), viscosity (n = 5), spreadability (n = 10) and syringability (n = 10) were evaluated, as well as the mechanical properties of hardness, cohesiveness, compressibility, elasticity, and adhesiveness using the texture profile assay (TPA) (n = 5) and ex vivo mucoadhesion (n = 5). The antibiofilm effect was assessed by counting colony-forming units (CFU/mL) (n = 8), metabolic activity (XTT) (n = 8), and scanning electron microscopy (SEM) (n = 1), using 0.12 % chlorhexidine gel (CG) as a positive control and G0 as a negative control. All groups demonstrated a neutral pH. G2 and G3 exhibited reduced viscosity and syringeability, along with increased spreadability. The incorporation of β-AgVO3 did not impact the gel's elasticity or cohesiveness but led to a decrease in hardness, compressibility, and adhesiveness. Ex vivo mucoadhesion was not altered by the addition of the nanomaterial. G3 showed a higher microbial reduction compared to GC and G0. It was concluded that the formulations with β-AgVO3 showed compatible physicochemical and mechanical properties for application and maintenance in the oral cavity. G3 showed superior antimicrobial effect against all microorganisms compared to GC and G0.

Combined application of earthworms and plant growth promoting rhizobacteria improve metal uptake, photosynthetic efficiency and modulate secondary metabolites levels under chromium metal toxicity in Brassica juncea L

Chromium (Cr) toxicity impairs essential morphological and metabolic activities in plants. The present investigation was carried out to evaluate the beneficial role of plant growth promoting rhizobacterial strains namely Pseudomonas aeruginosa (M1), Burkholderia gladioli (M2) and earthworms (Eisenia fetida) in alleviating Cr toxicity in 10 days old Brassica juncea L. The findings delineated that addition of earthworms and PGPR restored growth, boosted Cr uptake and showed upregulation of metal transporter genes (SULTR 1-4). Supplementation of rhizospheric amendments reinstated Cr induced impairment in photosynthetic attributes. Gaseous exchange attributes, the efficiency of PS II, the content of total phenols, anthocyanin and flavonoids was enhanced with application of earthworms along with PGPR. Confocal imaging of primary photosynthetic pigment (chlorophyll), accessory photosynthetic pigment (carotenoids) and total phenols showed maximum fluorescence with combined inoculation of earthworms and both microbial strains (M1M2). The gene expression analysis revealed that Phyotene synthase (PSY), Photosystem II core protein psb A, psb B were down regulated in Cr stressed seedlings which upon supplementation with earthworms and PGPR were upregulated. Further, Phenylalanine ammonialyase (PAL), chalcone synthase (CHS) were upregulated with addition of earthworms and PGPR. Increased nitric oxide content, enhanced activity and upregulation of nitrate reductase (NR) gene was observed with addition of PGPR and earthworms

Realizing UV driven microalgae photosynthesis to bio-fix CO2 by spectrum conversion

Microalgae, as efficient photosynthetic autotrophs, hold significant potential for carbon sequestration. However, their photosynthesis is constrained by the selective use of the solar spectrum. To enhance the utilization of natural light energy, a dual functional substrate with ultraviolet (UV) light conversion and microalgae biofilm support was proposed by coating a rare earth complexes phosphor agent onto transparent acrylic in this study. By doing this, UV light could directly drive microalgae biofilm photosynthesis by converting it to red light, resulting in a maximum 2.81 times increase in the carbon fixation rate. The UV conversion substrate increased the electron transport rate of microalgal photosystem II by 58.08%, effectively enhancing the photosynthetic rate and oxygen release, promoting the formation of a porous biofilm structure, and strengthening the material transfer within the biofilm. Additionally, reactive oxygen species (ROS) can cause oxidative stress, affecting cell growth and metabolic activity. With the use of the UV conversion substrate, the levels of ROS and malondialdehyde (MDA) in microalgae decreased by 84.56% and 42.35%, respectively. Consequently, UV-induced damage to microalgae cells was reduced, leading to a 52.52% increase in the photosynthesis rate and achieving UV driven photosynthetic carbon sequestration. This study demonstrated the UV conversion substrate’s effectiveness in reducing UV-induced damage in microalgae and enhancing photosystem activity, showing significant potential for carbon fixation within microalgal biofilms.

Taurine ameliorates radiation-induced oxidative stress in bone marrow mesenchymal stromal cells and promotes osteogenesis

Osteoradionecrosis of the jaw (ORNJ) is a severe complication following head and neck radiotherapy that significantly impacts the quality of life of patients. Currently, there is a lack of comprehensive understanding of the microenvironmental factors involved in ORNJ. In this study, we reveal the activation of taurine metabolism in irradiated mandibular stromal cells using scRNA-Seq and demonstrate a decrease in taurine levels in irradiated bone marrow mesenchymal stromal cells (BMSCs) through metabolomics. Compared with unirradiated BMSCs, taurine uptake in irradiated BMSCs increases. Taurine concentrations in the peripheral blood and jaws of irradiated mice are significantly lower than those in unirradiated mice (P = 0.0064 and 0.0249 respectively). Supplementation with taurine promotes osteogenic differentiation, reduces oxidative stress, and decreases DNA damage in irradiated BMSCs. Oral administration of taurine significantly improves the survival rate of irradiated mice and enhances osteogenesis in irradiated jaws. Our study highlights the role of taurine in the recovery from radiation-induced jaw injury, and suggests its potential as a non-invasive therapeutic option for combating ORNJ.

Study on the physiological responses and tolerance mechanisms to subchronic carbonate alkalinity exposure in the gills of Paramisgurnus dabryanus

Given the reduction of freshwater resources, saline-alkaline aquaculture has emerged as an effective approach to expand the fishery's accessible space. High carbonate alkalinity (CA) is a major stressor for aquatic organisms in saline-alkaline environments. Paramisgurnus dabryanus is a potential species for culture in saline-alkaline water, making it an ideal model for investigating the physiological responses and tolerance mechanisms to CA exposure in freshwater fishes. In the current study, P. dabryanus were exposed to 15 and 30mmol/L NaHCO3, combining blood biochemical, gill histological, transcriptomic, and metabolomic methods for conjoint analysis of response mechanisms. After 28-d exposure, the gill ventilation frequency of P. dabryanus decreased significantly, gill lamellae twisted and atrophied, and gill filament epithelial cells proliferated, potentially limiting gas exchange, whereas the accessory air-breathing frequency increased significantly, possibly for greater oxygen uptake. Serum osmolality and blood pH remained relatively steady, while serum ammonia levels rose significantly. A total of 3718 differentially expressed genes (DEGs) and 205 differential metabolites (DMs) were identified between the control group and 30mmol/L NaHCO3 group, involved in ion transport (Na+/K+-ATPase, V-type ATPase, carbonic anhydrase, and ABC transporters), ammonia transport (Rh glycoproteins and Aquaporins), amino acid metabolism, carbohydrate metabolism, and fatty acid metabolism. Furthermore, DEGs were significantly associated with cell-cell/ extracellular matrix interaction and protein synthesis. An integrated multi-omics analysis revealed the activation of carbon metabolism and TCA cycle. These results indicate that in response to CA exposure, P. dabryanus may facilitate carrier-mediated ion and ammonia transport to maintain the internal osmotic equilibrium and lessen the deleterious effects of blocked ammonia excretion. Meanwhile, amino acid metabolism and protein synthesis are disturbed, P. dabryanus can modulate carbohydrate catabolism to maintain energy homeostasis. The above findings provide novel insights into saline-alkaline adaptation in freshwater fishes, paving the way for future research and development of saline-alkaline-tolerant Cobitidae strains.

Metabolomics reveals changes in levels of fecal branched chain amino acids and organic acids in very preterm infants fed human milk fortified with bovine colostrum

Background & aimsHuman milk is the optimal diet for very preterm infants (VPIs), but it requires nutrient fortification to support growth. Bovine colostrum (BC), rich in intact proteins and bioactive components, could serve as a novel fortifier with potential benefits to VPIs gut health. To evaluate a possible effect of feeding BC on intestinal metabolism, the gut microbiota, and their interaction, we studied the fecal metabolome of VPIs in the first month of life, as compared with a conventional fortifier (CF, based on infant formula ingredients). MethodsFecal samples were collected from VPIs recruited to the FortiColos trial (NCT03537365, BC, n = 107; CF, n = 112) before (FT0) and one (FT1) or two (FT2) weeks after start of fortification and analyzed using 1H NMR spectroscopy. Abundances of metabolites were compared between BC versus CF groups. Further, temporal changes in metabolite levels after start of fortification, as well as correlations with specific gut bacterial genera were explored. ResultsInfants in the BC group had higher levels of fecal acetoacetate, choline, methanol, uracil, creatine, creatinine, lysine and a lower leucine at both FT1 and FT2, relative to the CF group. Asparagine, tryptophan and phenylalanine levels were higher, and butyrate was lower in the BC group at FT1. At FT2, higher fecal succinate and lower isoleucine were found in the BC group. In addition, eight metabolites (asparagine, phenylalanine, tryptophan, lysine, creatinine, acetoacetate, methanol and uracil) had fortification-specific changes over time. Correlations were found between succinate and unclassified Enterobacteriaceae, butyrate and Clostridium, uracil and Staphylococcus, as well as unclassified Enterobacteriaceae members. ConclusionOur study shows distinct fecal metabolome profiles in VPIs in the first weeks after fortification with BC versus CF. The fortification- and time-specific gut metabolite changes suggest that fortifiers influence luminal nutrient metabolism and microbiota activity in VPIs. Fortifier type for human milk affected gut health of VIPs via altered gut metabolite levels, interacting with microbiota in VPIs.

Associations between skeletal muscle phenotype, positional role, and on-ice performance in elite male ice hockey players.

We evaluated associations between muscle phenotype, positional role, and on-ice performance in male U20 Danish national team ice hockey players. Sixteen players (10 forwards, six defensemen) participated in a game with activity tracking. Resting thigh muscle biopsies were analyzed for metabolic enzyme activity and protein expression linked to performance. On-ice intermittent exercise capacity, repeated sprint ability, and maximal isometric knee-extensor torque were also assessed. No significant position-specific muscle phenotype characteristics were found, but forwards generally exhibited higher levels of several membrane proteins (p = 0.100-0.991). NAKα2, NAK∑, KATP, ClC-1, and NHE1 showed significant correlations with total distance (r = 0.52-0.59, p = 0.016-0.046), however, within positions these only persisted for KATP (r = 0.70, p = 0.024) and NAKα2 (r = 0.57, p = 0.085) in forwards, where CS enzyme activity also displayed a strong association with distance covered (r = 0.75, p = 0.019). For high-intensity skating, NAKα2 (r = 0.56, p = 0.025) and KATP (r = 0.50, p = 0.048) similarly exhibited the strongest associations, persisting within forwards (r = 0.63, p = 0.052 and r = 0.72; p = 0.018, respectively). In conclusion, although several muscle proteins involved in ion and metabolic regulation were associated with performance, only NAKα2 and KATP displayed consistent relationships within positions. Moreover, CS enzyme activity was strongly related to total distance within forwards, coherent with the proposed importance of oxidative capacity in intense intermittent exercise.

A genome-wide investigation of the mechanism underlying the effect of exogenous boron application on sugar content and overall quality of “Benihoppe” strawberries

In recent years, the widespread application of growth regulators and nutrients to boost yield and quality of strawberry fruits has led to the rapid growth of strawberry industry globally. Although the effects of major nutrients on strawberry yield have been widely studied, investigations into the effect of trace elements such as boron remain limited. This study examined the effect of boron application on the yield and quality of “Benihoppe” strawberry fruits. Nutrient solutions with varying boron concentrations (0, 0.024, 0.048, 0.072, and 0.096 mM) were applied to the plants, and their effect on fruit quality was evaluated. The results indicated that boron application enhanced the yield per plant, nutrient composition (total amino acid and vitamin C content), antioxidant properties (total phenol) and volatile components (esters) in strawberry fruits. Specifically, treatment with 0.048 mM boron concentration significantly increased the accumulation of soluble sugars, such as sucrose, whose concentration was 154.29% higher than that of the control treated with 0 mM concentration. This enhancement is attributable to the regulated expression of sucrose phosphate synthase (maker-Fvb2-2-augustus-gene-229.38) and β-fructofuranosidase-1/2/3 (augustus-masked-Fvb5-4-processed-gene-2.0, maker-Fvb5-3-augustus-gene-272.30, and maker-Fvb5-1-augustus-gene-0.37) genes, which play crucial roles in sugar metabolism and enzyme activity. Overall, boron application enhanced the quality of “Benihoppe” strawberries. The findings of this study offer substantial theoretical and practical guidance for using boron fertilizers in strawberry farming.

Enhanced bioremediation of bensulfuron-methyl contaminated soil by Hansschlegelia zhihuaiae S113: metabolic pathways and bacterial community structure

Bensulfuron-methyl (BSM), a widely used herbicide, can persist in soil and damag sensitive crops. Microbial degradation, supplemented with exogenous additives, provides an effective strategy to enhance BSM breakdown. Hansschlegelia zhihuaiae S113 has been shown to efficiently degrade this sulfonylurea herbicide. However, depending solely on a single strain for degradation proves inefficient and unlikely to achieve ideal remediation in practical applications. This study assessed the impact of various carbon sources on the degradation efficiency of S113 in BSM-polluted soil. Among these, glucose was the most effective, achieving a 98.7% degradation rate after 9 d of inoculation. In addition, seven intermediates were detected during BSM degradation in soil through the cleavage of the phenyl ring ester bond, the pyrimidine rings, and urea bridge peptide bond, among other pathways. 2-amino-4,6-dimethoxy pyrimidine (ADMP), and 2-(aminosulfonylmethyl)-methyl benzoate(MSMB) were the primary intermediates. These metabolites were less toxic to maize, sorghum, and bacteria than the BSM. Community structure analysis indicated that variations in exogenous carbon sources and environmental pollutants significantly improved the ecological functions of soil microbial communities, enhancing pollutant degradation. Addition of carbon sources notably affected soil microbial community structure, modifying metabolic activities and interaction patterns. Specifically, glucose substantially increased the richness and diversity of soil bacterial communities. These findings offer valuable insights for field remediation practices and contributed to the development of more robuste soil pollution management strategies.

Hepatic enzyme induction and its potential effect on thyroid hormone metabolism in the metamorphosing tadpole of Xenopus laevis (African clawed frog).

Hepatic enzyme induction, an inherent defense system against xenobiotics, is known to simultaneously affect endocrine system functions in mammals under specific conditions, particularly thyroid hormone (TH) regulation. While this phenomenon has been studied extensively, the pathway leading to this indirect thyroid effect in mammals has unclear applicability to amphibians, despite the importance of amphibian species in assessing thyroid-disruptive chemicals. Here, we investigated the effects of three well-known mammalian enzyme inducers-β-naphthoflavone (BNF), pregnenolone carbonitrile (PCN), and sodium phenobarbital (NaPB)-on the gene expression of phase-I and phase-II metabolizing enzymes in Xenopus laevis tadpoles. Waterborne exposure to BNF and PCN significantly induced the expression of both phase-I (cytochrome P450, CYP) and phase-II enzymes (UDP-glucuronosyltransferase, UGT and sulfotransferase, SULT), but in different patterns, while NaPB exposure induced CYP2B expression without affecting phase-II enzymes in tadpoles, in contrast to mammals. Furthermore, an ex vivo hepatic enzyme activity assay confirmed that BNF treatment significantly increased phase-II metabolic activity (glucuronidation and sulfation) toward TH. These results suggest the potential for certain mammalian enzyme inducers to influence TH clearance in X. laevis tadpoles. Our findings provide insights into the profiles of xenosensing activity and enzyme induction in amphibians, which can facilitate a better understanding of the mechanisms of indirect effects on the thyroid system via hepatic enzyme induction in nonmammalian species.

Metabolic activation of WHO-congeners PCB28, 52, and 101 by human CYP2A6: evidence from in vitro and in vivo experiments.

Despite extensive research on the metabolism of polychlorinated biphenyls (PCBs), knowledge gaps persist regarding their isoform-specific biotransformation pathways. This study aimed to elucidate the role of different cytochrome P450 enzymes in PCB metabolism, focusing on WHO-congeners 2,4,4'-trichlorobiphenyl (PCB28), 2,2',5,5'-tetrachlorobiphenyl (PCB52), and 2,2',4,5,5'-pentachlorobiphenyl (PCB101). Utilizing engineered HEK293 cell lines, we investigated the in vitro metabolism of these PCBs by CYP1A2, CYP2C8, CYP2C9, CYP3A4, CYP2A6, and CYP2E1, revealing robust production of hydroxylated metabolites. Our results show that CYP2A6 plays a major role in the metabolism of these congeners responsible for predominant formation of para-position hydroxylated metabolites, with concentrations reaching up to 1.61µg/L (5,89nM) for PCB28, 316.98µg/L (1,03µM) for PCB52, and 151.1µg/L (441nM) for PCB101 from a 20µM parent PCB concentration. Moreover, concentration-dependent cytotoxic and cytostatic effects induced by reactive intermediates of the PCB hydroxylation pathway were observed in HEK293CYP2A6 cells, for all three congeners tested. CYP2A6 was specifically capable of activating PCBs 28 and 101 to genotoxic metabolites which produced genetic defects which were propagated to subsequent generations, potentially contributing to carcinogenesis. In a clinical study examining CYP2A6 enzyme activity in formerly exposed individuals with elevated internal PCB levels, a participant with increased enzyme activity showed a direct association between the phenotypic activity of CYP2A6 and the metabolism of PCB28, confirming the role of CYP2A6 in the in vivo metabolism of PCB28 also in humans. These results altogether reinforce the concept that CYP2A6 plays a pivotal role in PCB congener metabolism and suggest its significance in human health, particularly in the metabolism of lower chlorinated, volatile PCB congeners.

Outside-in enhancement of phosphate solubilizing bacteria by sludge biochar for phenol remediation in soil: Pollution stress reduction, electron transfer gain and secretion regulation

The high toxicity of phenol poses a significant barrier to bacterial bio-removal capacity. This study innovatively proposes the utilization of sludge biochar (SB) to embed phosphate-solubilizing bacteria (CZB1), creating a composite system (SB-CZB1) that enhances the bio-removal efficiency of phenol. The research findings indicate that after being embedded with biochar, Phosphorus solubilizing bacteria (PSB) achieved a significant enhancement (49.7 %-67.0 %) in phenol removal efficiency across different concentration gradients (1500, 2000, 2500 mg/L). SB component in the SB-CZB1 composite system provides a stable and favorable growth environment for CZB1, significantly enhancing microbial metabolic activity. Notably, it stimulates CZB1 to secrete succinic acid and malic acid (with increases of 3.8 % and 23.9 %, respectively), effectively mitigating the detrimental effects of phenol toxicity on microorganisms. Three-dimensional fluorescence and electrochemical analysis demonstrate that SB not only exhibits exceptional electrochemical performance but also stimulates CZB1 to generate redox-active substances (quinone components in humic acid). The electron transfer rates between microorganisms and phenol in the SB-CZB1 system are 9.75–21.71 times and 23.94–63.95 times higher than those in SB and CZB1 alone, respectively. Meanwhile, the abundant oxygen-containing functional groups (COH/COC, COOH, CO) on the surface of SB-CZB1 play a pivotal role in the adsorption and removal of phenol. Furthermore, pot experiments further reveal that the SB-CZB1 composite system significantly enhances the removal efficiency of phenol in soil (98.1 %). Additionally, the application of SB-CZB1 effectively modulated soil properties, notably increasing available potassium and available phosphorus content by 40.86 % and 136.69 %. This application not only enriched soil microbial diversity but also promoted the proliferation of native organic pollutant remediation bacteria in the soil. Notably, the proliferation of Bacillus species affiliated with CZB1 was the most significant, increasing by 82.1 %. This result confirms that biochar embedding technology effectively enhanced the colonization of CZB1 in the soil, thereby significantly accelerating the bio-removal process of phenol.

Silk fibroin nanoparticles for locoregional cancer therapy: Preliminary biodistribution in a murine model and microfluidic GMP-like production

Silk fibroin nanoparticles (SFNs) have been widely investigated for drug delivery, but their clinical application still faces technical (large-scale and GMP-compliant manufacturing), economic (cost-effectiveness in comparison to other polymer-based nanoparticles), and biological (biodistribution assessments) challenges. To address biodistribution challenge, we provide a straightforward desolvation method (in acetone) to produce homogeneous SFNs incorporating increasing amounts of Fe2O3 (SFNs-Fe), detectable by Magnetic Resonance Imaging (MRI), and loaded with curcumin as a model lipophilic drug. SFNs-Fe were characterized by a homogeneous distribution of the combined materials and showed an actual Fe2O3 loading close to the theoretical one. The amount of Fe2O3 incorporated affected the physical-chemical properties of SFNs-Fe, such as polymer matrix compactness, mean diameter and drug release mechanism. All formulations were cytocompatible; curcumin encapsulation mitigated its cytotoxicity, and iron oxide incorporation did not impact cell metabolic activity but affected cellular uptake in vitro. SFNs-Fe proved optimal for biodistribution studies, as MRI showed significant nanoparticle retention at the administration site, supporting their potential for locoregional cancer therapy. Finally, technical and economic challenges in SFN production were overcome using a GMP-compliant microfluidic scalable technology, which optimized preparation to produce smaller particle sizes compared to manual methods and reduced acetone usage, thus offering environmental and economic benefits. Moreover, enabling large-scale production of GMP-like SFNs, this represents a considerable step forward for their application in the clinic.

The synergistic interaction of fungi with different structural characteristics improves the leaching of lithium from lepidolite

The aim of this study was to explore the improvement of lithium leaching from lepidolite by microbial co-culture, focusing on the synergistic effect of different structural fungi in improving the leaching performance of biological systems. The results showed that in the single leaching experiment, the multicellular fungi and the unicellular yeast showed weak effects. Multicellular fungus is limited by insufficient EPS secretion, while unicellular yeast is non-mycelial organisms with weak acid production capacity and limited effect on minerals. However, in the combined leaching experiment, the interspecific collaboration promoted the synthetic and metabolic activity of the two strains, resulting in changes in the type and content of organic acids, polysaccharides, proteins and humus. The content of citric acid reached the highest value of 16.98g·L-1 at about 22 d, and the EPS secreted by unicellular yeast promoted the mycelium adhesion and mineral wrapping of multicellular fungi. The combined action of the two enhanced the effects of acidification, complexation and mycelium destruction, and improved the leaching effect. This study revealed the synergistic metabolic mechanism of lepidolite leaching by fungi with different structures, verified the effectiveness of microbial co-culture to improve the leaching rate, and provided a basis for industrial application. In addition, the use of co-culture technology has a positive impact on commercial production and environmental protection.

Are we better together? Addressing a combined treatment of pitavastatin and temozolomide for brain cancer

Pitavastatin is commonly prescribed to treat hypercholesterolemia through the regulation of cholesterol biosynthesis. Interestingly, it has also demonstrated a great potential for treating brain tumors, although the detailed cytotoxic mechanism, particularly in glioblastoma, remains incompletely understood. This work explores the activity of pitavastatin in 2D and 3D glioblastoma models, in an attempt to provide a more representative and robust overview of its anticancer potential in glioblastoma. The results show that not only is pitavastatin 10-1000 times-fold more effective in reducing tumoral metabolic activity than temozolomide, but also demonstrate a synergistic activity with this alkylating drug. In addition, low micromolar concentrations of this statin strongly impair the growth and the invasion ability of multicellular tumor spheroids. The obtained qRT-PCR and proteomics data highlight the modulation of cell death via apoptosis (BAX/BCL2, CASP9) and autophagy (BECN1, BNIP3, BNIP3L and LC3B), as well as an epithelial to mesenchymal transition blockage (HTRA1, SERPINE1, WNT5A, ALDH3B1 and EPHA2) and remodeling of the extracellular matrix (VCAN, SERPINE1 and TGFBI). Overall, these results lay the foundation for further investigations on the potential combinatory clinical treatment with temozolomide.

Efficient removal of nitrate and ammonium by strain Pseudomonas poae EH-E3 under acidic pH and low-temperature conditions

The combined effect of low temperature and an acidic environment may restrict the growth and metabolic activity of microorganisms involved in nitrogen removal. To overcome poor biological nitrogen removal efficiency during low-temperature treatment of acidic wastewater, a novel pure strain of Pseudomonas poae EH-E3 was successfully isolated and screened based on its strong heterotrophic nitrification and aerobic denitrification capacity. Strain EH-E3 could completely eliminate inorganic nitrogen sources of nitrate within 30h and ammonium within 20h at 15 °C and pH 5.0, with corresponding removal efficiencies for total nitrogen of 95.13% and 91.35%, respectively. Nitrogen balance testing indicated that strain EH-E3 removed 43.25% of ammonium, 45.72% of nitrate, and 51.20% of nitrite in the form of gaseous products. Enzyme activity assays revealed that the specific activities of ammonia monooxygenase, nitrate reductase, and nitrite reductase were 0.533, 0.956, and 0.011 U/mg proteins, respectively. These findings suggest that strain EH-E3 has strong potential for use in low-temperature treatment of acidic wastewater.