Search Results (1,322)

Unspecific peroxygenases (UPOs) are promising biocatalysts for oxyfunctionalizations in future sustainable economies and can be efficiently immobilized on the cell surface of their heterologous production yeast. This immobilization has versatile uses, ranging from the mL to m³ scale; but the production of the yeast surface displayed UPOs, and their handling has yet to be optimized to advance sustainable industrial processes in light of the UN’s sustainable development goals. Here, we present optimized production protocols for surface-displayed UPOs for shaken and stirred systems in different scales and describe suitable storage conditions and a sterilization method. We utilized one-factor-at-a-time and design of experiments approaches. We were able to streamline published protocols for shaken flask cultivations to achieve a 60% increase in volumetric activity, using reduced amounts of media. We also show at least a doubling of final activity for bioreactor cultivations by utilizing a different medium than the industry standard. Finally, we present a novel, robust protocol for parallelized methanol-induced enzyme production in Komagataella phaffii in a BioLector XT^® reactor. Enzyme activity did not decrease and even increased by our recommended sterilization method and during storage over 87 days. This study aims to advance the yeast surface display immobilization method by providing methods for efficient production, storage and utilization of this promising biocatalyst. Full article

Background/Objectives: Ovarian cancer (OC) remains one of the most lethal malignancies of the female reproductive system. Glucose oxidase (GOx) has emerged as a potential therapeutic agent in cancer treatment by inducing tumor starvation through glucose depletion. Nonetheless, its clinical application is constrained due to its systemic toxicity, immunogenicity, poor in vivo stability, and short half-life. These challenges can be addressed through nanotechnology; in particular, biogenic mesoporous silica nanoparticles (MSNs) offer promise as drug delivery systems (DDSs) that enhance therapeutic efficacy while minimizing side effects. Methods: Biogenic MSNs were extracted from the Equisetum myriochaetum plant via acid digestion, functionalized with 3-aminopropiltrietoxysilane (APTES) and glutaraldehyde (GTA), and loaded with GOx. The free and immobilized MSNs were characterized using FTIR, DLS, XRD, SEM/EDX, and BET techniques. A colorimetric approach was employed to quantify the enzymatic activity of both the free and immobilized GOx. The MTT assay was employed to assess the viability of SKOV3 cells. The obtained IC₅₀ concentration of the nanoformulation was administered to SKOV3 cells to analyze the expression of cancer-related genes using RT-qPCR. Results: IC₅₀ values of 60.77 ng/mL and 111.6 µg/mL were ascertained for the free and immobilized GOx, respectively. Moreover, a significant downregulation of the oncogene β-catenin (CTNNB1) was detected after 24 h with the nanoformulation. Conclusions: Our findings indicate that GOx-loaded biogenic MSNs may serve as a potential therapeutic agent for ovarian cancer. This is, to the best of our knowledge, the first report exploring the effect of GOx-loaded biogenic MSNs on SKOV3 cells. Full article

Lavender essential oil (LEO) was analyzed using gas chromatography coupled with a mass selective detector (GC-MS), detecting linalool and linalyl acetate as its major constituents. The biological activity of the LEO was evaluated in vitro using a normal mouse fibroblast cell line (L929), where it showed no cytotoxic effects. To assess its therapeutic effect in vivo, a broiler chicken model (Ross 308) was employed. Birds were divided into three groups: the control group (C) without any hydrogel supplementation; the H group, supplemented with alginate hydrogel capsules without LEO; and the HE groups, which received hydrogel capsules containing immobilized LEO. Capsules were provided on chick paper for voluntary intake from day 1 to day 10. At the end of the production cycle, the cecum was dissected and preserved for subsequent molecular analyses. Results demonstrated that dietary supplementation with alginate hydrogel containing immobilized LEO (HE group) positively influenced the production parameters and intestinal health in broiler chickens. Dietary supplementation with alginate hydrogel-encapsulated LEO exerts therapeutic effects in broilers. Full article

Globally, phosphogypsum (PG) is the primary by-product of the phosphorus industry. Aspergillus niger (A. niger), one of the most powerful types of phosphate-solubilizing fungi (PSF), can secrete organic acids to dissolve the phosphates in PG. This study investigated heavy metal (HM) remediation by PG and A. niger under the co-existence of Pb and Cd. It demonstrated that 1 mmol/L Pb²⁺ stimulated the bioactivity of A. niger during incubation, based on the CO₂ emission rate. PG successfully functioned as P source for the fungus, and promoted the growth of the fungal cells. Meanwhile, it also provided sulfates to immobilize Pb in the solution. The subsequently generated anglesite was confirmed using SEM imaging. The immobilization rate of Pb reached over 95%. Under co-existence, Pb²⁺ and 0.01 mmol/L Cd²⁺ maximized the stimulating effect of A. niger. However, the biotoxicity of Pb²⁺ and elevated Cd²⁺ (0.1 mmol/L) counterbalanced the stimulating effect. Finally, 1 mmol/L Cd²⁺ dramatically reduced the fungal activity. In addition, organic matters from the debris of A. niger could still bind Pb²⁺ and Cd²⁺ according to the significantly lowered water-soluble Pb and Cd concentrations. In all treatments with the addition of Cd²⁺, the relatively high biotoxicity of Cd²⁺ induced A. niger to absorb more Pb²⁺ to minimize the sorption of Cd²⁺ based on the XRD results. The functional group analysis of ATR-IR also confirmed the phenomenon. This pathway maintained the stability of Pb²⁺ immobilization using the fungus and PG. This study, hence, shed light on the application of A. niger and solid waste PG to remediate the pollution of Pb and Cd. Full article

To promote environmentally sustainable remediation and resource recovery from ion-adsorption rare earth tailings (IRET), this study comprehensively investigated the previously isolated strain REO-01 by examining its sulfate-reducing performance, Cd(II) immobilization potential, and physiological and biochemical responses under varying environmental conditions. Strain REO-01 was identified as a Gram-negative facultative anaerobe with strong sulfate-reducing activity and effective Cd(II) immobilization capacity. During a 96 h incubation period, the strain entered the exponential growth phase within 36 h, after which the OD₆₀₀ values plateaued. Concurrently, the culture pH increased from 6.83 to 7.5, and the oxidation-reduction potential (ORP) declined to approximately −300 mV. Cd(II) concentrations decreased from 0.2 mM to 3.33 μM, corresponding to a removal efficiency exceeding 95%, while sulfate concentrations declined from 1500 mg/L to 640 mg/L, with a maximum reduction efficiency of 66.16%. The strain showed optimal growth at 25–40 °C and near-neutral pH (6–7), whereas elevated Cd(II) concentrations (≥0.2 mM) significantly inhibited cell growth. A sulfate concentration of 1500 mg/L was found to be optimal for cellular activity. Among the tested carbon sources, sodium lactate at 4.67 g/L yielded the most favorable results, reducing ORP to −325 mV, increasing pH to 7.6, and lowering Cd(II) and sulfate concentrations to 3.33 μM and 510 mg/L, respectively. These findings highlight the strong potential of strain REO-01 for simultaneous sulfate reduction and Cd(II) remediation, supporting its application in the in situ bioremediation and resource utilization of rare earth tailings. Full article

In polymer-based lithium batteries, polymer electrolytes (PEs) exhibit limited ionic conductivity at room temperature (25 °C). To address this issue, this paper describes a hexagonal-structure-based single-ion conducting gel polymer electrolyte (h-SICGPE) framework with a robust and efficient cross-linked polymer network, applicable to polymer-based batteries even at 25 °C. The proposed cross-linked polymer network backbone of the h-SICGPE, as a semisolid-state thin film type, has the homogeneous honeycomb structure incorporating anion receptor(s) inside each of its hexagonal closed cells and is obtained by cross-linking between trimethylolpropane tris(3-mercaptopropionate) and poly(ethylene glycol) diacrylate in a newly synthesized anion–receptor solution. The excellent structural capability of the h-SICGPE incorporating Li⁺/TFSI⁻ can enhance ionic conductivity and electrochemical stability by suppressing crystallinity and expanding free volume. Further, the anion receptor in its free volume helps to effectively increase the lithium-ion transference number by immobilizing counter-anions. Experimental results demonstrate dramatically superior performance at 25 °C, such as ionic conductivity (2.46 mS cm⁻¹), oxidative stability (4.9 V vs. Li/Li⁺), coulombic efficiency (97.65%), and capacity retention (88.3%). These results confirm the developed h-SICGPE as a promising polymer electrolyte for high-performance polymer-based lithium batteries operable at 25 °C. Full article

Sediments are key players in the optimum functioning of ecosystems; however, they also represent the largest known repository of harmful contaminants. The vast variety of these sediment-associated contaminants may exert harmful effects on marine communities and can impair ecosystem functioning. Whole-cell biosensors are a rapid and biologically relevant tool for assessing environmental toxicity. Therefore, in this study, we developed a bioassay-based toxicity measurement system using genetically modified bacteria to create a whole-cell optical biosensor. Briefly, reporter bacteria were integrated and immobilized using a calcium alginate matrix on fiber-optic tips connected to a photon counter placed inside a light-proof, portable case. The calcium alginate matrix acts as a semi-permeable membrane that protects the reporter-encapsulated optical fiber tips and allows the inward passage of toxicant(s) to induce a dose-dependent response in the bioreporter. The samples were tested by directly submerging the fiber tip with immobilized bacteria into vials containing either water or suspended sediment samples, and the subsequent bioluminescent responses were acquired. In addition to bioavailable sediment toxicity assessments, conventional chemical methods, such as liquid chromatography–mass spectroscopy (LC-MS) and inductively coupled plasma optical emission spectroscopy (ICP-OES), were used for comprehensive evaluation. The results demonstrated the efficacy of the biosensor in detecting various toxicity levels corresponding to identified contaminants, highlighting its potential integration into environmental monitoring frameworks for enhanced sediment and water quality assessments. Despite its utility, this study notes the system’s operational challenges in field conditions, recommending future enhancements for improved portability and usability in remote locations. Full article

The low effectiveness of various brain cancer treatment methods is due to a number of significant challenges. Most of them are unable to penetrate the blood–brain barrier (BBB) when drugs are administered systemically through the bloodstream. Nanoscale particles play a special role among materials capable of binding drug molecules and successfully crossing the BBB. Biopolymeric nanoparticles (NPs) demonstrate excellent biocompatibility and have the remarkable ability to modify the environment surrounding tumor cells, thereby potentially improving cellular uptake of delivery agents. In our research, nanoscale polyelectrolyte complexes (PECs) ranging in size from 56 to 209 nm were synthesized by ionic interaction of the oppositely charged polysaccharides pectin and chitosan. The structural characteristics of these complexes were carefully characterized by infrared (FTIR) and Raman spectroscopy. The immobilization efficiency of antitumor drugs was comprehensively evaluated using UV spectrophotometry. The cytotoxicity of the NPs was evaluated in the U87-MG cell line. The preliminary data indicate a significant decrease in the metabolic activity of these tumor cells. Important details on the interaction of the NPs with an endothelial layer structurally similar to the BBB were obtained by simulating the BBB using a model based on human blood vessels. Our studies allowed us to establish a significant correlation between the kinetic parameters of drug immobilization and the ratio of biopolymer concentrations in the initial compositions, which provides valuable information for future optimization of drug delivery system design. Full article

Protein phosphorylation is one of the most common and important post-translational modifications (PTMs) and is highly involved in various biological processes. Ideal adsorbents with high sensitivity and specificity toward phosphopeptides with large coverage are therefore essential for enrichment and mass spectroscopy-based phosphoproteomics analysis. In this study, a newly designed IMAC adsorbent composite was constructed on the graphene matrix coated with mesoporous silica. The outer functional 3D-network layer was prepared by free radical polymerization of the phosphonate-functionalized vinyl imidazolium salt monomer and subsequent metal immobilization. Due to its unique structural feature and high content of Ti⁴⁺ ions, the resulting phosphonate-immobilized adsorbent composite G@mSiO₂@PPFIL-Ti⁴⁺ exhibits excellent performance in phosphopeptide enrichment with a low detection limit (0.1 fmol, tryptic β-casein digest) and superior selectivity (molar ratio of 1:15,000, digest mixture of β-casein and bovine serum albumin). G@mSiO₂@PPFIL-Ti⁴⁺ displays high tolerance to loading and elution conditions and thus can be reused without a marked decrease in enrichment efficacy. The captured phosphopeptides can be released globally, and mono-/multi-phosphopeptides can be isolated stepwise by gradient elution. When applying this material to enrich phosphopeptides from human lung cell lysates, a total of 3268 unique phosphopeptides were identified, corresponding to 1293 phosphoproteins. Furthermore, 2698 phosphorylated peptides were found to be differentially expressed (p < 0.05) between human lung adenocarcinoma cells (SPC-A1) and human normal epithelial cells (Beas-2B), of which 1592 were upregulated and 1106 were downregulated in the cancer group. These results demonstrate the material’s superior enrichment efficiency in complex biological samples. Full article

Today, there is considerable interest in creating artificial microbial consortia to solve various biotechnological problems. The use of such consortia allows for the improvement of process indicators, namely, increasing the rate of accumulation of target products and enhancing the conversion efficiency of the original substrates. In this work, the prospects for creating artificial consortia based on anaerobic sludge (AS) with cells of different yeasts were confirmed to increase the efficiency of methanogenesis in glucose- and glycerol-containing media and obtain biogas with an increased methane content. Yeasts of the genera Saccharomyces, Candida, Kluyveromyces, and Pachysolen were used to create the artificial consortia. Their concentration in the biomass of consortium cells was 1.5%. Yeast cells were used in an immobilized form, which was obtained by incorporating cells into a cryogel of polyvinyl alcohol. The possibility of increasing the efficiency of methanogenesis by 1.5 times in relation to the control (AS without the addition of yeast cells) was demonstrated. Using a consortium composed of methanogenic sludge and yeast cells of the genus Pachysolen, known for their ability to convert glycerol into ethanol under aerobic conditions, the possibility of highly efficient anaerobic conversion of glycerol into biogas was shown for the first time. Analysis of the metabolic activity of the consortia not only for the main components of the gas phase (CH₄, CO₂, and H₂) and metabolites in the cell culture medium, but also for the concentration of intracellular adenosine triphosphate (ATP), controlled by the method of bioluminescent ATP-metry, showed a high level of functionality and thus, prospects for using such consortia in methanogenesis processes. The advantages and the prospect of using the developed consortia instead of individual AS for the treatment of methanogenic wastewater were confirmed during static tests conducted with several samples of real and model waste. Full article

The transition toward sustainable and decentralized energy solutions necessitates the development of innovative bioelectronic systems capable of harvesting and converting renewable energy. Here, we present a novel photo-bioelectrochemical fuel cell architecture based on a biohybrid anode integrating laser-induced graphene (LIG), poly(3,4-ethylenedioxythiophene) (PEDOT), and isolated thylakoid membranes. LIG provided a porous, conductive scaffold, while PEDOT enhanced electrode compatibility, electrical conductivity, and operational stability. Compared to MXene-based systems that involve complex, multi-step synthesis, PEDOT offers a cost-effective and scalable alternative for bioelectrode fabrication. Thylakoid membranes were immobilized onto the PEDOT-modified LIG surface to enable light-driven electron generation. Electrochemical characterization revealed enhanced redox activity following PEDOT modification and stable photocurrent generation under light illumination, achieving a photocurrent density of approximately 18 µA cm⁻². The assembled photo-bioelectrochemical fuel cell employing a gas diffusion platinum cathode demonstrated an open-circuit voltage of 0.57 V and a peak power density of 36 µW cm⁻² in 0.1 M citrate buffer (pH 5.5) under light conditions. Furthermore, the integration of a charge pump circuit successfully boosted the harvested voltage to drive a low-power light-emitting diode, showcasing the practical viability of the system. This work highlights the potential of combining biological photosystems with conductive nanomaterials for the development of self-powered, light-driven bioelectronic devices. Full article

Metals are natural components of the lithosphere, whose amounts and bioavailability are increasing in many areas due to their continuous release from both natural sources and intensive human activities. Some metals are essential or beneficial for living organisms, while others are non-essential and potentially toxic. When present at higher concentrations, even essential and beneficial metal ions can become harmful to all forms of life. Bacteria, unicellular organisms that have been exposed to metals since the earliest stages of life on Earth, have evolved metabolic pathways involving essential metals as well as diverse strategies to cope with metal toxicity. In the domain Bacteria, two main strategies have been identified: (i) metal exclusion, which includes cell wall sequestration and immobilization of metals in extracellular exopolysaccharides, siderophores, and other soluble microbial products, as well as (ii) metal tolerance, involving intracellular sequestration of metals (e.g., by metallothioneins, or low molecular weight thiols) as well as enzymatic conversion of metals to less toxic forms and/or its active efflux. Microorganisms possessing such adaptive traits are considered valuable agents for potential application in medicine, environmental sciences, and bioengineering (e.g., bioremediation and/or biomining). Full article

Background/Objectives: Lung cancer is the leading cause of cancer-related mortality worldwide. Accurate radiotherapy (RT) planning alongside chemotherapy and immunotherapy is critical for improving treatment outcomes for inoperable non-metastatic cases. Conventional computed tomography (CT)-based planning may be inadequate for accurately identifying tumor margins and the location of nodal disease. We investigated whether ¹⁸F-labeled fluorodeoxyglucose positron emission tomography (¹⁸F-FDG PET-CT) imaging can assist in target volume delineation for primary, nodal, and metastatic disease in the RT planning and overall therapeutic planning of patients. Methods: In this single-center, prospective study, we recruited 34 patients with histologically confirmed locally advanced non-small-cell lung carcinoma (NSCLC). All patients underwent ¹⁸F-FDG PET-CT-based RT simulation. Two sequential RT plans were created by the same radiation oncologist: one based on CT alone and the other PET-CT. Planning target volumes (PTVs) and PET-CT-guided adjustments were analyzed to assess their impact. Standardized protocols for immobilization, imaging, target delineation, and dose prescription were applied. Results: A total of 34 patients (31 males and 3 females) were recruited in the study. ¹⁸F-FDG PET-CT detected distant metastases in 7/34 (20.6%) patients, altering the overall therapeutic plan in 4/34 (11.8%) and allowing radical RT in 3 of them who had oligometastatic disease (8.8%). It modified RT planning in 26/34 (76.5%) patients and clarified malignancy in atelectatic areas. Nodal involvement was identified in 3/34 patients (8.8%) and excluded in 3/34 cases, avoiding unnecessary nodal irradiation. Additional involved nodes were revealed in 12/34 (35.3%) patients, requiring dose escalation. Overall, changes to the tumor PTV were made in 23/30 (76.6%) and to the nodal PTV in 19/30 (63.3%) cases (p < 0.0001). Primary tumor and nodal PTVs increased in 20/34 (66.7%) and 13/34 (43.3%), respectively. Conclusions: ¹⁸F-FDG PET-CT significantly improves RT planning by more precisely defining tumor and nodal volumes, identifying undetected lesions, and guiding dose adaptation. Larger long-term studies are required to confirm potential locoregional control and survival improvements. Full article

There is evidence that calcitriol is the only biologically active vitamin D metabolite. This review summarizes data on the regulation of renal and extrarenal synthesis of calcitriol by nutritional, physiologic, mechanical, genetic, and disease-related factors. Relatively low circulating calcitriol due to low substrate availability, i.e., low circulating 25-hydroxyvitamin D, has been reported in nutritional rickets, osteomalacia, obesity, and preeclampsia. In these situations, vitamin D supplementation can increase circulating calcitriol and, together with calcium, prevent rickets/osteomalacia and reduce the risk of preeclampsia and obesity-related type 2 diabetes mellitus. However, the correction of low circulating calcitriol due to mechanical unloading/immobilization by vitamin D supplementation is not effective in preventing osteoporotic fractures. Circulating calcitriol is also low in diseases such as cardiac and renal failure. Both illnesses share some other similarities regarding dysregulated calcium/phosphate metabolism, including elevated parathyroid hormone and fibroblast growth factor-23, suggesting similar treatment strategies. Genetic disorders of vitamin D metabolism are rare and can affect circulating calcitriol differently. Calcitriol synthesis in immune cells is obviously not primarily dependent on circulating 25-hydroxyvitamin D, which challenges the use of vitamin D for infection prevention. Since various factors can differently influence calcitriol regulation, more personalized preventive/therapeutic strategies of targeting calcitriol synthesis are necessary. Full article

Seven wild-type lactic acid bacteria, belonging to Lactiplantibacillus plantarum and Lactococcus cremoris species, were isolated from beetroots and white mushrooms and evaluated for their safety and functional profile. Lc. cremoris isolates were sensitive to all antibiotics tested, while L. plantarum strains exhibited resistance in certain antibiotics. Among them, Lc. cremoris FBMS_5810 showed the highest cholesterol removal ability (51.89%) and adhesion capacity to Caco-2 cell lines (32.14%), while all plant origin strains exhibited strong antagonistic and inhibitory activity against foodborne pathogens, as well as high survival potential during an in vitro digestion model. Subsequently, freeze-dried immobilized Lc. cremoris FBMS_5810 cells on oat flakes were prepared with initial cell loads >8.5 log CFU/g, and the effect of trehalose as a cryoprotectant in cell viability during storage at room and refrigerated temperatures for up to 180 days was studied. A significant reduction in cell loads was observed in all cases studied. However, freeze-dried immobilized Lc. cremoris FBMS_5810 cells on oat flakes prepared using trehalose as a cryoprotectant stored at 4 °C exhibited the highest cell viability (8.75 log CFU/g) after 180 days. In the next step, functional breakfast cereals enriched with freeze-dried immobilized Lc. cremoris FBMS_5810 cells on oat flakes (produced with (MLT) or without (ML) trehalose) were developed and stored at room and refrigerated temperatures for 180 days. The initial cell levels ≥ 9.18 log CFU/g were achieved, while a significant decrease was recorded during storage in all cases. The maintenance of cell loads ≥ 7.75 log CFU/g was documented in the case of both ML and MLT samples stored at 4 °C; however, the presence of trehalose in MLT samples resulted in cell viability 7.52 log CFU/g after 180 days of storage at room temperature. Importantly, the functional breakfast cereals were accepted by the panel during the sensory evaluation. Full article