Search Results (21)

With the deepening of oil and gas exploration and development into ultra-deep and ultra-high pressure environments, the pressure of wellhead equipment is becoming higher and higher. The sealing performance of the casing head hanger is directly related to the safety and reliability of the whole wellhead equipment. Firstly, based on the numerical simulation method, the sealing performance of three different metal seal rings—H-type, X-type, and U-type—under 175 MPa working conditions is compared and analyzed. The simulation results show that the sealing performance of the H-type metal sealing ring is better than that of the X-type and U-type. The parametric analysis method is further used to study the influence of the structural parameters of the convex radius and the bottom angle of the H-ring on its sealing performance. The results show that when the convex radius is designed to be 3 mm, and the bottom angle is 90°, the effective contact width reaches 5.91 mm, and the contact uniformity is the best. Finally, based on the H-type metal sealing ring sample trial-produced with optimized parameters, a 175 MPa nitrogen medium sealing pressure test was completed on an 8 1/8” all-metal sealed mandrel casing hanger. The test results show that the system pressure drop is 0.7 MPa during the 5-min pressure stabilization process, which has good sealing reliability. Full article

Chitosan oligosaccharides (COSs) with defined degrees of polymerization (DP) exhibit distinct bioactivities with promising applications in food, pharmaceutical, and agricultural industries. However, the specific and sustainable production of COSs remains challenging due to the broad product distribution of wild-type chitosanases and the difficulties in enzyme recovery and reuse. In this study, we employed rational design to engineer a GH46 chitosanase (CsnB) from Bacillus sp. BY01 for chitobiose production. Through homology modeling and molecular docking analysis, 15 mutants were designed by targeting key residues structurally critical for substrate stabilization, product release, and active-site geometry in the substrate-binding subsites. The D78Y mutant exhibited exclusive specificity for chitobiose, demonstrating a specific activity of 102.4 U/mg and yielding chitobiose with a purity exceeding 98%, thereby surpassing the previously reported enzymes for chitobiose production. To address the challenges of enzyme stability, purification costs, and product separation, we developed a ReELP system by integrating elastin-like polypeptides (ELPs) with a ReverseCatcher/ReverseTag peptide pair. This system enabled one-step purification and co-immobilization of CsnB-D78Y directly from cell lysate onto biomimetic silica nanoparticles, achieving 96.8% immobilization efficiency and 90.7% activity recovery. The immobilized enzyme exhibited enhanced thermal and pH stability, retaining approximately 50% activity after 12 h at 40 °C compared to only 5.7% for the free enzyme. In reusability assays, the immobilized CsnB-D78Y maintained efficient chitobiose production over 5 consecutive cycles. This work provides a green and cost-effective strategy for the specific and sustainable production of chitobiose, offering new insights into enzyme engineering and immobilization for industrial COS production. Full article

Tunnels serve as a critical hub in urban transportation networks; their monotonous and enclosed environment is prone to inducing speeding behavior, necessitating an efficient vehicle speed monitoring system. Traditional methods suffer from high costs and slow response times, making them inadequate for the complex scenarios encountered in tunnel environments. This study proposes a real-time tunnel vehicle speed monitoring system based on YOLOv8s and DeepSORT. YOLOv8s is used to detect and classify cars, trucks, and buses, while DeepSORT applies Kalman filtering and the Hungarian algorithm to construct motion trajectories. Vehicle speed is estimated through perspective geometric transformation combined with a sliding-window approach, with a speeding threshold of 100 km/h and corresponding visual alerts. Using surveillance video from an expressway tunnel as the dataset, the system achieved detection accuracies of 98% for cars, 96% for trucks, and 91% for buses. Speed detection performance metrics included an average speed deviation (ASD) of 2.54 km/h, a deviation degree of vehicle speed (DDVS) of 3.12, vehicle speed stability (VST) of 1.22, and speed difference ratio (SDR) of 2.9%. Analysis revealed a longitudinal “deceleration–acceleration–deceleration” inverted U-shaped speed profile along the tunnel. Statistical tests confirmed these findings: the Mann–Whitney U test showed highly significant differences in vehicle speeds between cars and trucks across different tunnel sections, and the Kruskal–Wallis test further indicated significant speed variations across the entrance, middle, and exit segments for both vehicle types. Full article

Small-scale wind turbines are becoming increasingly important in renewable energy systems due to their ability to operate in low-wind-speed environments and adapt to various installation locations, especially in areas with energy shortages. This paper presents the design, analysis and development of a Helical Vertical Axis type Wind Turbine (H-VAWT) using uPVC pipe as the blade material, offering a lightweight, low-cost, and corrosion resistant solution. The blade structure is optimized for use in residential and off-grid areas with unstable wind conditions. Structural analysis is conducted in ANSYS, including static load analysis (deformation, equivalent stress, shear stress, maximum stress), torsional and bending stress, and modal analysis to assess mechanical performance and vibrational stability. Three blade designs are initially considered, and the helical model (0–45° twist) is selected based on simulation results. The prototype is successfully fabricated and tested under different wind speeds, showing effective power generation, with favorable results in power output, power coefficient, tip-speed ratio (TSR), and relative velocity. Full article

The purpose of this paper is to investigate a class of novel symmetric coupled fuzzy fractional partial differential equation system involving the Caputo–Katugampola (C-K) generalized Hukuhara (gH) derivative. Within the framework of C-K gH differentiability, two types of gH weak solutions are defined, and their existence is rigorously established through explicit constructions via employing Schauder fixed point theorem, overcoming the limitations of traditional Lipschitz conditions and thereby extending applicability to non-smooth and nonlinear systems commonly encountered in practice. A typical numerical example with potential applications is proposed to verify the existence results of the solutions for the symmetric coupled system. Furthermore, we introduce Ulam–Hyers stability (U-HS) theory into the analysis of such symmetric coupled systems and establish explicit stability criteria. U-HS ensures the existence of approximate solutions close to the exact solution under small perturbations, and thereby guarantees the reliability and robustness of the systems, while it prevents significant deviations in system dynamics caused by minor disturbances. We not only enrich the theoretical framework of fuzzy fractional calculus by extending the class of solvable systems and supplementing stability analysis, but also provide a practical mathematical tool for investigating complex interconnected systems characterized by uncertainty, memory effects, and spatial dynamics. Full article

The aim of this study is to evaluate the mechanical properties and long-term stability of 3D-printable resins for permanent fixed dental prostheses (FDPs), focusing on whether material performance is influenced by 3D-printer type or by differences in resin formulations. Specimens (N = 621) were printed. CAD/CAM blocks (BRILLIANT Crios) served as control. Flexural strength (FS) with elastic modulus (E_calc), Weibull modulus (m), Martens’ hardness (HM), indentation modulus (EIT), elastic modulus (E_RFDA), shear modulus (G_RFDA), and Poisson’s Ratio (ν) were measured initially, after water storage (24 h, 37 °C), and after thermocycling (5–55 °C, 10,000×). SEM analysis assessed microstructure. Data were analyzed using Kolmogorov–Smirnov, ANOVA with Scheffe post hoc, Kruskal–Wallis with Mann–Whitney U, and Weibull statistics with maximum likelihood (α = 0.05). A ceramic crown printed with Midas showed higher FS, HM, and EIT values after thermocycling than with Pro55s, and higher E_calc scores across all aging regimes. A Varseo Smile Crown Plus printed with VarseoXS and AsigaMax showed a higher FS value than TrixPrint2, while AsigaMax achieved the highest initial E_calc and E_RFDA values, and VarseoXS did so after thermocycling. HM, EIT, and G_RFDA were higher for TrixPrint2 and AsigaMax printed specimens, while ν varied by system and aging. 3Delta Crown, printed with AsigaMax, showed the highest FS, E_calc, HM, EIT, and m values after aging. VarseoSmile triniQ and Bridgetec showed the highest E_RFDA and G_RFDA values depending on aging, and Varseo Smile Crown Plus exhibited higher ν initially and post-aging. Printer system and resin formulation significantly influence the mechanical and aging behaviors of 3D-printed FDP materials, underscoring the importance of informed material and printer selection to ensure long-term clinical success. Full article

Isocitrate dehydrogenase 1 (IDH1) mutations are key drivers of glioma biology, influencing tumor aggressiveness and treatment response. To elucidate their molecular impact, we performed proteome analysis on patient-derived (PD) and U87MG glioma cell models with either mutant or wild-type IDH1. We quantified over 6000 protein groups per model, identifying 1594 differentially expressed proteins in PD-AS (IDH1_MUT) vs. PD-GB (IDH1_WT) and 904 in U87_MUT vs. U87_WT. Both IDH1_MUT models exhibited enhanced MHC antigen presentation and interferon signaling, indicative of an altered immune microenvironment. However, metabolic alterations were model-dependent: PD-AS cells shifted toward glycolysis and purine salvage, while U87_MUT cells retained oxidative phosphorylation, potentially due to D2-hydroxyglutarate (2OHG)-mediated HIF1A stabilization. We also observed a predominance of downregulated DNA repair proteins in IDH1_MUT models, particularly those involved in homologous recombination. In contrast, RB1 and ASMTL were strongly upregulated in both IDH1_MUT models, implicating them in DNA repair and cellular stress responses. We also found distinct expression patterns of proteins regulating histone methylation in IDH1_MUT cells, favoring increased methylation of H3K4, H3K9, and H3K36. A key driver of this may be the upregulation of SETD2 in PD-AS, an H3K4 and H3K36 trimethyltransferase linked to the recruitment of HIF1A as well as DNA mismatch repair proteins. This study uncovers candidate biomarkers and pathways relevant to glioma progression and therapeutic targeting, but also underscores the complexity of predicting glioma pathogenesis and treatment responses based on IDH1 mutation status. While proteome profiling provides valuable insights, a comprehensive understanding of IDH1_MUT gliomas will likely require integrative multi-omics approaches, including DNA/RNA methylation profiling, histone and protein post-translational modification analyses, and targeted DNA damage and repair assays. Full article

The underlying structural features of the mismatch between catalytic and cytostatic properties in L-asparaginase from Rhodospirillum rubrum (RrA) and three of its mutants were investigated. The rationale for selecting the specific mutations (RrA_{A64V, E67K}; RrA_{R118H, G120R}; RrA_{E149R, V150P, F151T}) is to elucidate the role of inter-subunit interaction in RrA and its impact on catalytic efficiency and stability. Bioinformatic modeling revealed a predominantly negative surface charge on RrA with limited positive charge clusters in the vicinity of the interface region. Thus, some negatively charged groups were replaced with positively charged ones to enhance the electrostatic interactions and stabilize the enzyme quaternary structure. RrA_{A64V, E67K} and RrA_{R118H, G120R} additionally contained an N-terminal 17-amino acid capsid peptide derived from the bacteriophage T7 (MASMTGGQQMGRGSSRQ), which could potentially affect the conformational stability of theenzymes. Circular dichroism (CD) spectroscopy was applied to the kinetic parameters analysis of Asn hydrolysis and showed that native RrA displayed a V_max of 30 U/mg and a K_M of 4.5 ± 0.5 mM. RrA_{E149R, V150P, and F151T} exhibited a substantially increased V_max of 57 U/mg. The catalytic efficiency of V_max/K_M also improved compared to the native enzyme: the V_max/K_M increased from approximately 7 U/mg × mM⁻¹ (for the native enzyme) to 9 U/mg × mM⁻¹ for Mut3. Other mutants exhibited less pronounced changes. Thermo-denaturation studies allowed us to determine the phase transition parameters of the RrA variants in comparison with commercial reference sample EcA. RrA_{A64V, E67K} and RrA_{R118H, G120R} exhibited the most favorable phase transition parameters, with melting temperatures (T_m) of 60.3 °C and 59.4 °C, respectively, exceeding that of the wild-type RrA (54.6 °C) and RrA_{E149R, V150P, F151T} (52 °C). The EcA demonstrated a slightly superior thermal stability, with a T_m of 62 °C. The mutations showed a significant effect on protein stability during trypsinolysis. Therefore, RrA_{E149R, V150P, F151T} showed higher resistance (45% activity remaining after 30 min of trypsin exposure) compared to the native RrA retained 20% activity. EcA preparations exhibited lower stability to trypsinolysis (losing over 90% activity in 15 min). The cytostatic effects were evaluated using MTT assays against K562 (leukemic) and A549 (lung carcinoma) cell lines. The MTT assays with K562 cells revealed that RrA_{E149R, V150P, F151T} (IC₅₀ of 10 U/mL) and RrA_{R118H, G120R} (IC₅₀ of 11.5 U/mL) exhibited superior antiproliferative activity compared to native enzymes RrA (IC₅₀ of 15 U/mL) and EcA (24 U/mL). RrA_{E149R, V150P, F151T} showed the most significant improvement in cytostatic activity. The results obtained indicate that the substitutions in RrA_{E149R, V150P, F151T} resulted in the improvement of the enzyme biocatalytic properties and an increase in the resistance to aggregation and trypsinolysis. This highlights the role of electrostatic interactions in stabilizing the oligomeric structure of the enzyme, which eventually translates into an improvement in cytostatic efficiency and antiproliferative forces. Full article

Alginate lyase degrades alginate through the β-elimination mechanism to produce alginate oligosaccharides (AOS) with notable biochemical properties and diverse biological activities. However, its poor thermostability limits large-scale industrial production. In this study, we employed a rational computational design strategy combining computer-aided evolutionary coupling analysis and ΔΔG_fold evaluation to enhance both the thermostability and catalytic activity of the alginate lyase VxAly7B-CM. Among ten single-point mutants, the E188N and S204G mutants exhibited increases in T_m from 47.0 °C to 48.9 °C and 50.2 °C, respectively, with specific activities of 3701.02 U/mg and 2812.01 U/mg at 45 °C. Notably, the combinatorial mutant E188N/S204G demonstrated a ΔT_m of 5 °C and an optimal reaction temperature up to 50 °C, where its specific activity reached 3823.80 U/mg—a 31% increase. Moreover, its half-life at 50 °C was 38.4 h, which is 7.0 times that of the wild-type enzyme. Protein structural analysis and molecular dynamics simulations suggested that the enhanced catalytic performance and thermostability of the E188N/S204G mutant may be attributed to optimized surface charge distribution, strengthened hydrophobic interactions, and increased tertiary structure stability. Overall, our findings provided valuable insights into enzyme stabilization strategies and supported the industrial production of functional AOS. Full article

In recent decades, many researchers have pointed out that derivatives and integrals of the non-integer order are well suited for describing various real-world materials, for example, polymers. It has also been shown that fractional-order mathematical models are more effective than integer-order mathematical models. Thereby, given these considerations, the investigation of qualitative properties, in particular, Ulam-type stabilities of fractional differential equations, fractional integral equations, etc., has now become a highly attractive subject for mathematicians, as this represents an important field of study due to their extensive applications in various branches of aerodynamics, biology, chemistry, the electrodynamics of complex media, polymer science, physics, rheology, and so on. Meanwhile, the qualitative concepts called Ulam–Hyers–Mittag-Leffler (U-H-M-L) stability and Ulam–Hyers–Mittag-Leffler–Rassias (U-H-M-L-R) stability are well-suited for describing the characteristics of fractional Ulam-type stabilities. The Banach contraction principle is a fundamental tool in nonlinear analysis, with numerous applications in operational equations, fractal theory, optimization theory, and various other fields. In this study, we consider a nonlinear fractional Volterra integral equation (FrVIE). The nonlinear terms in the FrVIE contain multiple variable delays. We prove the U-H-M-L stability and U-H-M-L-R stability of the FrVIE on a finite interval. Throughout this article, new sufficient conditions are obtained via six new results with regard to the U-H-M-L stability or the U-H-M-L-R stability of the FrVIE. The proofs depend on Banach’s fixed-point theorem, as well as the Chebyshev and Bielecki norms. In the particular case of the FrVIE, an example is delivered to illustrate U-H-M-L stability. Full article

Bacterial laccase exhibits substantial application potential in various fields. In this study, we constructed a mutation library of CotA laccase from Bacillus pumilus using error-prone PCR, and we performed four rounds of enrichment screening under malachite green (MG) pressure. The results demonstrated that the proportions of the four selected mutant strains were significantly increased. The enzyme activities of the four final mutants PW2, PW5, PW4G, and PW6 were 94.34, 75.74, 100.66, and 87.04 U/mg, respectively, representing a significant increase of approximately 2- to 3-fold compared to the wild-type CotA laccase. Notably, PW4 exhibited significantly improved thermal stability at 90 °C and pH tolerance at pH 12.0. Homology modeling analysis revealed that alterations in the amino acid sequence rendered the spatial structure of the enzyme’s catalytic site more favorable for substrate binding. For instance, the substitution of T262A in PW2 and V426I in PW4 shortened the side chains of the amino acids, thereby enlarging the substrate-binding cavity. The G382D mutation in PW2 and PW5 may induce altered protein conformation via spatial steric hindrance or electrostatic interactions, consequently impacting enzyme activity and stability. These findings provide valuable insights for enhancing the industrial application of bacterial laccase. Full article

Chitosanases play a significant part in the hydrolysis of chitosan to form chitooligosaccharides (COS) that possess diverse biological activities. This study aimed to enhance the productivity of Paenibacillus elgii TKU051 chitosanase by fermentation from chitinous fishery wastes. The ideal parameters for achieving maximum chitosanase activity were determined: a squid pens powder amount of 5.278% (w/v), an initial pH value of 8.93, an incubation temperature of 38 °C, and an incubation duration of 5.73 days. The resulting chitosanase activity of the culture medium was 2.023 U/mL. A chitosanase with a molecular weight of 25 kDa was isolated from the culture medium of P. elgii TKU051 and was biochemically characterized. Liquid chromatography with tandem mass spectrometry analysis revealed that P. elgii TKU051 chitosanase exhibited a maximum amino acid identity of 43% with a chitosanase of Bacillus circulans belonging to the glycoside hydrolase (GH) family 46. P. elgii TKU051 chitosanase demonstrated optimal activity at pH 5.5 while displaying remarkable stability within the pH range of 5.0 to 9.0. The enzyme displayed maximum efficiency at 60 °C and demonstrated considerable stability at temperatures ≤40 °C. The presence of Mn²⁺ positively affected the activity of the enzyme, while the presence of Cu²⁺ had a negative effect. Thin-layer chromatography analysis demonstrated that P. elgii TKU051 chitosanase exhibited an endo-type cleavage pattern and hydrolyzed chitosan with 98% degree of deacetylation to yield (GlcN)₂ and (GlcN)₃. The enzymatic properties of P. elgii TKU051 chitosanase render it a promising candidate for application in the production of COS. Full article

This paper is devoted to generalizing the standard system of Navier boundary value problems to a fractional system of coupled sequential Navier boundary value problems by using terms of the Caputo derivatives. In other words, for the first time, we design a multi-term fractional coupled system of Navier equations under the fractional boundary conditions. The existence theory is studied regarding solutions of the given coupled sequential Navier boundary problems via the Krasnoselskii’s fixed-point theorem on two nonlinear operators. Moreover, the Banach contraction principle is applied to investigate the uniqueness of solution. We then focus on the Hyers–Ulam-type stability of its solution. Furthermore, the approximate solutions of the proposed coupled fractional sequential Navier system are obtained via the generalized differential transform method. Lastly, the results of this research are supported by giving simulated examples. Full article

Analytical and numerical investigations were performed to study the influence of the Soret and Dufour effects on double-diffusive convection in a vertical porous layer filled with a binary mixture and subject to horizontal thermal and solute gradients. In particular, the study was focused on the effect of Soret and Dufour diffusion on bifurcation types from the rest state toward steady convective state, and then toward oscillatory convective state. The Brinkman-extended Darcy model and the Boussinesq approximation were employed to model the convective flow within the porous layer. Following past laboratory experiments, the investigations dealt with the particular situation where the solutal and thermal buoyancy forces were equal but acting in opposite direction to favor the possible occurrence of the rest state condition. For this situation, the onset of convection could be either supercritical or subcritical and occurred at given thresholds and following various bifurcation routes. The analytical investigation was based on the parallel flow approximation, which was valid only for a tall porous layer. A numerical linear stability analysis of the diffusive and convective states was performed on the basis of the finite element method. The thresholds of supercritical, $R_{TC}^{sup}$, and overstable, $R_{TC}^{over}$, convection were computed. In addition, the stability of the established convective flow, predicted by the parallel flow approximation, was studied numerically to predict the onset of Hopf’s bifurcation, $R_{TC}^{Hopf}$, which marked the transition point from steady toward unsteady convective flows; a route towards the chaos. To support the analytical analyses of the convective flows and the numerical stability methodology and results, nonlinear numerical solutions of the full governing equations were obtained using a second-order finite difference method. Overall, the Soret and Dufour effects were seen to affect significantly the thresholds of stationary, overstable and oscillatory convection. The Hopf bifurcation was marked by secondary convective flows consisting of superposed vertical layers of opposite traveling waves. A good agreement was found between the predictions of the parallel flow approximation, the numerical solution and the linear stability results. Full article

In this work hydrogen bonding in a diverse set of 36 unnatural and the three natural Watson Crick base pairs adenine (A)–thymine (T), adenine (A)–uracil (U) and guanine (G)–cytosine (C) was assessed utilizing local vibrational force constants derived from the local mode analysis, originally introduced by Konkoli and Cremer as a unique bond strength measure based on vibrational spectroscopy. The local mode analysis was complemented by the topological analysis of the electronic density and the natural bond orbital analysis. The most interesting findings of our study are that (i) hydrogen bonding in Watson Crick base pairs is not exceptionally strong and (ii) the N–H⋯N is the most favorable hydrogen bond in both unnatural and natural base pairs while O–H⋯N/O bonds are the less favorable in unnatural base pairs and not found at all in natural base pairs. In addition, the important role of non-classical C–H⋯N/O bonds for the stabilization of base pairs was revealed, especially the role of C–H⋯O bonds in Watson Crick base pairs. Hydrogen bonding in Watson Crick base pairs modeled in the DNA via a QM/MM approach showed that the DNA environment increases the strength of the central N–H⋯N bond and the C–H⋯O bonds, and at the same time decreases the strength of the N–H⋯O bond. However, the general trends observed in the gas phase calculations remain unchanged. The new methodology presented and tested in this work provides the bioengineering community with an efficient design tool to assess and predict the type and strength of hydrogen bonding in artificial base pairs. Full article