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The reusability of enzymes is a fundamental aspect of sustainable biotechnology and the development of biosensors. This study presents one of the first quantitative evaluations of DNA polymerase reusability by utilizing integrated quartz crystal microbalance (QCM) kinetics and real-time monitoring of exonuclease activity. The results showed that immobilized T7 DNA polymerase retained approximately 50% of its initial activity after three 90-min cycles and around 20% after five cycles. Significantly lower activities were observed for shorter, 45-min cycles. This indicates an unexpected time-dependent enhancement in stability for longer reaction times. The findings suggest a promising trend in enzyme stability and reusability, establishing a quantitative relationship between reaction duration and enzyme performance. This relationship offers a scalable pathway for the regeneration of biosensors and for sustainable enzymatic catalysis. Additionally, the work provides a transferable framework that can be applied to other DNA-processing enzymes, which supports long-term biosensor performance and industrial biocatalysis. The demonstrated approach offers a transferable and scalable methodology for the development of reusable polymerase-based biosensors and sustainable biocatalytic systems.

Energy storage investments are essential for achieving sustainable development goals; however, their high costs and long-term returns create significant uncertainty for investors. As a result, investment decisions are shaped not only by rational evaluations but also by psychological and emotional factors. Despite this reality, behavioral economics-based variables remain largely underexplored in the energy storage literature. To address this gap, this study identifies the most critical behavioral determinants influencing the performance of energy storage investments. A novel decision support system is developed for G7 countries by integrating z-scoring, criteria impact multiplicative assessment (CIMAS), interval-valued spherical fuzzy numbers, and the ranking of alternatives through functional mapping of criterion sub-intervals into a single interval (RAFSI). The proposed framework enhances the representation of uncertainty and reduces individual biases through statistically derived expert weighting. The results indicate that risk perception and overconfidence are the most influential behavioral factors, while Germany and the United States exhibit the strongest behavioral economics-based performance in energy storage investments.

Purpose This study aimed to evaluate the biomechanical response or load transfer on the osteoporotic L1 vertebra under torsional loading. Methods. To achieve this goal, a numerical model of osteoporotic vertebra in various trabecular bone degenerations was developed and tested. The mechanical behavior of the model was represented taking into account the anisotropic properties of the cancellous bone, which provided a more realistic mechanical picture of the biological subsystem. To ensure the reliability of osteoporotic degradation, the thinning of cortical bone and the appearance of gaps between trabecular bone and cortical bone were also taken into account when creating the models. Results. Finite element (FE) analysis showed that the deformations of cortical bone thinning and detachment of the cortical bone from the trabecular tissue lead to local instability of the vertebrae. As a result, the cortical bone of a vertebra loses its load-bearing capacity, even if the strength limit is not reached. Conclusions. The results obtained allow us to state that taking into account the thinning of the trabeculae, which creates voids, is extremely important for load-bearing capacity of osteoporotic vertebrae. However, a limitation of this study is the lack of experimental data to ensure consistency with the computer simulation results.

The United Nations presented a global action plan of an integrated nature for sustainable development, and the role of cities is seen as central in efforts to implement these goals. Integrating city performance evaluation methods containing sustainability indicators is becoming increasingly popular. The United Nations and other international organizations and researchers analyze urban sustainability in the context of different countries. However, holistic assessment methods that can show if and to what extent a city is sustainable, measure its progress, and provide information for policy development are only just emerging. Our Synergies and Trade-offs Recommendation Method for Sustainable Cities and Communities (the STRICT method) proposes that country and city metrics are connected in multifaceted ways through tradeoffs and synergies. As a result of this analysis, city models and maps were created, and digital evidence-based recommendations were provided to urban and national policymakers based on the findings of the analysis. A 1% improvement in country success individual metrics led to a synergistic effect in 189 out of 219 city indicators (86.3%). The STRICT method suggests a notable impact of a country’s social, cultural, and political success on the city indicators.

Novel composite materials have been prepared by primary paper sludge (PS) and biopolymer (modified corn starch (MCS). The MCS is used as a binder. The aim of the research was to investigate the properties, microstructure and durability (water erosion) of composite materials (CM). The CM were formed from 100 wt% of paper sludge and 20 wt% of various concentrations (2.5%, 5.0% and 7.5%) of MCS were added. Corn starch was heated 3 h at 150 °C temperature. The CM produced had a bulk density of 0.89–1.18 g/cm3, compressive strength at 10% relative deformation 4.8–9.3 MPa, thermal conductivity 0.092–0.132 W/m·K. Additionally, this study underscores that the CM show good durability (resistance to water erosion) and hygrothermal qualities. In fact, the MSC solution incorporated in the paper sludge showed improved physical-mechanical properties and durability of the CM. The research shows that CM can be formed from paper sludge and MCS, and these can be used as indoor masonry units because they also have good mechanical, hydroscopic and thermal insulation properties.

Purpose: Poor lung cancer patients’ outcomes and survival rates demand the discovery of new biomarkers for the specific, significant, and less invasive detection of non-small cell lung cancer (NSCLC) progression. The present study aimed to investigate the potential of miRNA expression as biomarkers in NSCLC utilizing a preclinical cell culture setup based on screening of miRNAs in NSCLC cells grown in 3D cell culture. Patients and Methods: The study was performed using lung cancer cell lines, varying in different levels of aggressiveness: NCI-H1299, A549, Calu-1, and NCI-H23, as well as noncancerous bronchial epithelial cell line HBEC3, which were grown in 3D cell culture. Total RNA from all cell lines was extracted and small RNA libraries were prepared and sequenced using the Illumina NGS platform. The expression of 8 differentially expressed miRNAs was further validated in 89 paired tissue specimens and plasma samples obtained from NSCLC patients. Statistical analysis was performed to determine whether miRNA expression and clinicopathological characteristics of NSCLC patients could be considered as independent factors significantly influencing PFS or OS. Results: Differentially expressed miRNAs, including let-7d-3p, miR-10a-3p, miR-28-3p, miR-28-5p, miR-100-3p, miR-182-5p, miR-190a-5p, and miR-340-5p, were identified through next-generation sequencing in NSCLC cell lines with varying levels of aggressiveness. Validation of patient samples, including tumor and plasma specimens, revealed that out of the 8 investigated miRNAs, only plasma miR-10a-3p showed a significant increase, which was associated with significantly extended progression-free survival (PFS) (p=0.009). Furthermore, miR-10a-3p in plasma emerged as a statistically significant prognostic variable for NSCLC patients’ PFS (HR: 0.5, 95% CI: 0.3– 0.9, p=0.029). Conclusion: Our findings of screening miRNA expression patterns in NSCLC cells grown in 3D cell culture indicated that the expression level of circulating miR-10a-3p has the potential as a novel non-invasive biomarker to reflect the short-term prognosis of NSCLC patients.

BACKGROUND: Healthy children’s gait support patterns play a critical role in their development and overall well-being. Therefore, in order to develop a correct gait, it is necessary to constantly update knowledge. OBJECTIVE: To identify differences in gait support among children in neighbouring countries. METHODS: 44 healthy children from Poland and Lithuania (4–11 years old) participated in the study. The spatiotemporal and plantar pressure parameters of 88 neutrally aligned feet were analysed and compared. RESULTS: Statistically significant differences between stance, single-limb support, double support, swing duration, cadence, and velocity, max. force and pressure in the forefoot, as well as in the times of occurrence of max. forces in all three zones. Defined that age is related (p< 0.05) to cadence (R= 0.32), swing phase (R= 0.53), max. force under the midfoot (R= 0.35) and the heel (R= 0.47), max. pressure under the forefoot (R=−0.52), midfoot (R=−0.63) and heel (R=−0.47). CONCLUSION: The results can help caregivers, as well as clinicians and researchers, understand how gait mechanics change with development and the growth course of the children of that country. Also, the results are important for the analysis and comparison of children’s gait, as control reference data from the same country.

This paper analyses the dynamic behaviour of centrifugal milk separator's bearing-rotor system equipped with vibration absorbers. It is shown that rotational milk motion inside the separator causes a centrifugal force that increases dynamic unbalance of the rotor and has a negative effect on performance of the bearings. Mathematical simulation of bearing-rotor system has shown that the rotor rotates at frequency that is outside the resonant frequency region and the amplitude resonances are associated with the bearings and the components of step-up gear. Investigations of vibration transmissibility of vibration damping system have shown that the resonant frequencies are outside the rotor resonance interval and damping system suppresses the effect of separator with milk on rotor bearings and housing. However, it distorts results of diagnostic measurements. Experimental studies confirmed the last statement and showed that results of indirect (through the housing) vibration measurements cannot be used to diagnose the state of damped bearing-rotor system.

Graphene oxide (GO) are novel nanomaterials with a wide range of applications due to their high absorption capacity. This study was undertaken with a view to assess the bioaccumulation and acute toxicity of GO used in combination with the heavy metal mixture (Cr, Cu, Ni and Zn) to fish embryos and larvae. For this purpose, Salmo trutta embryos and larvae were subjected to the 4-day long treatment with three different concentrations of GO, the metal mixture, which was prepared of four metals at the concentrations corresponding to the maximum-permissible-concentrations for EU inland waters (Cr-0.01, Cu-0.01, Ni-0.034, and Zn-0.1 mg/L), and with GO in combination with MIX (GO+MIX). When used in combination with the metal mixture, GO exhibited a high metal sorption capacity. The obtained confocal fluorescence microscopy results showed that GO located in the embryo chorion causing its damage; in larvae, however, GO were found only in the gill region. Results of these experiments confirmed the hypothesis that GO affects the accumulation of metals and mitigates their toxic effects on organism. In embryos, the acute toxicity of exposure to GO and co-exposure to MIX+GO was found to manifest itself through the decreased heart rate (HR) and malondialdehyde (MDA) level and through the increased metallothionein (MT) concentration. Meanwhile, in larvae, GO and MIX+GO were found to induce genotoxicity effects. However, changes in HR, MDA, MT, gill ventilation frequency, yolk sack absorption and cytotoxicity compared with those of the control group were not recorded in larvae. The obtained results confirmed our hypothesis: the combined effect of MIX and GO was less toxic to larvae (especially survival) than individual effects of MIX components. However, our results emphasize that fish exposure to GO alone and in combination with heavy metal contaminants (MIX+GO) even at environmentally relevant concentrations causes health risks that cannot be ignored.

Pulsed electric fields (PEFs) are commonly used to facilitate the delivery of various molecules, including pharmaceuticals, into living cells. However, the applied protocols still require optimization regarding the conditions of the permeabilization process, i.e., pulse waveform, voltage, duration, and the number of pulses in a burst. This study highlights the importance of electrochemical processes involved in the electropermeabilization process, known as electroporation. This research investigated the effects of electroporation on human non-small cell lung cancer cells (A549) in potassium (SKM) and HEPES-based buffers (SHM) using sub-microsecond and microsecond range pulses. The experiments were performed using 100 ns – 100 μs (0.6–15 kV/cm) bursts with 8 pulses in a sequence. It was shown that depending on the buffer composition, the susceptibility of cells to PEF varies, while calcium enhances the cytotoxic effects of PEF, if high cell membrane permeabilization is triggered. It was also determined that electroporation with calcium ions induces oxidative stress in cells, including lipid peroxidation (LPO), generation of reactive oxygen species (ROS), and neutral lipid droplets. Here, we demonstrated that calcium ions and optimized pulse parameters could potentiate PEF efficacy and oxidative alternations in lung cancer cells. Thus, the anticancer efficacy of PEF in lung cancers in combination with standard cytostatic drugs or calcium ions should be considered, but this issue still requires in-depth detailed studies with in vivo models.