Search Results (90)

Electroporation-mediated gene delivery is a cornerstone of synthetic biology, offering several advantages over other methods: higher efficiencies, broader applicability, and simpler sample preparation. Yet, electroporation protocols are often challenging to integrate into highly multiplexed workflows, owing to limitations in their scalability and tunability. These challenges ultimately increase the time and cost per transformation. As a result, rapidly screening genetic libraries, exploring combinatorial designs, or optimizing electroporation parameters requires extensive iterations, consuming large quantities of expensive custom-made DNA and cell lines or primary cells. To address these limitations, we have developed a High-Throughput Microfluidic Electroporation (HTME) platform that includes a 384-well electroporation plate (E-Plate) and control electronics capable of rapidly electroporating all wells in under a minute with individual control of each well. Fabricated using scalable and cost-effective printed-circuit-board (PCB) technology, the E-Plate significantly reduces consumable costs and reagent consumption by operating on nano to microliter volumes. Furthermore, individually addressable wells facilitate rapid exploration of large sets of experimental conditions to optimize electroporation for different cell types and plasmid concentrations/types. Use of the standard 384-well footprint makes the platform easily integrable into automated workflows, thereby enabling end-to-end automation. We demonstrate transformation of E. coli with pUC19 to validate the HTME’s core functionality, achieving at least a single colony forming unit in more than 99% of wells and confirming the platform’s ability to rapidly perform hundreds of electroporations with customizable conditions. This work highlights the HTME’s potential to significantly accelerate synthetic biology Design-Build-Test-Learn (DBTL) cycles by mitigating the transformation/transfection bottleneck. Full article

Background and Objective: Bladder cancer (BC) is the ninth most common malignancy worldwide. Squamous cell carcinoma (SqCC), a rare histological variant, accounts for approximately 2–5% of all BC cases. Compared to urothelial carcinoma, the predominant subtype, research on SqCC remains limited and shows inconsistent findings regarding prognosis. This study aimed to compare survival outcomes between patients with SqCC and those with pure urothelial carcinoma (PUC). Methods: This retrospective, observational study analyzed pathology reports from 2549 transurethral resections of bladder tumors and 632 cystectomies performed at our institution between 1 December 2010 and 31 December 2023. Following pathological re-evaluation, 33 patients with SqCC and 132 with PUC were identified. After 1:3 propensity score matching, 20 patients with SqCC and 58 with PUC were included in the final analysis. Demographic, clinicopathological features, and survival outcomes were compared between groups. Results: The median follow-up was 2.31 years (range: 0.17–13.50). No significant differences in baseline demographic or clinical characteristics were observed, except for the type of surgery. Kaplan–Meier analysis demonstrated no significant differences in disease-free survival (DFS; p = 0.961) or overall survival (OS; p = 0.847) between SqCC and PUC groups. Multivariate Cox regression analysis identified T stage, nodal involvement, and adjuvant chemotherapy (CT) as independent predictors of DFS, while sex and metastasis at diagnosis were significant predictors of OS. Conclusion: Survival outcomes (DFS and OS) did not significantly differ between patients with SqCC and patients with PUC. Prognosis was more closely associated with disease stage at diagnosis, sex, and adjuvant CT. Further large-scale studies are warranted. Full article

Background/Objective: Iatrogenic ureteral injury is a rare but serious complication of colorectal cancer surgery. Although prophylactic ureteral catheterization (PUC) is used to facilitate intraoperative ureter identification and reduce the risk of ureteral injury, its efficacy is debated. We aimed to evaluate the clinical utility and outcomes of PUC in colorectal cancer surgery. Methods: This retrospective study included 42 patients who underwent PUC before colorectal cancer surgery at the Tohoku University Hospital between February 2010 and September 2024. Preoperative ureteral stents were inserted via cystoscopy under general anesthesia. Patient demographics, surgical techniques, indications for catheterization, and post-procedural complications were reviewed. Results: PUC was most frequently performed in patients with left-sided colorectal cancer (61.9%) and local recurrence of rectal cancer (31%). Ureteral catheterization was indicated in patients with a history of pelvic surgery (47.6%) or tumor proximity to the ureter (26.2%). Open surgery was performed in 90.5% of the cases, whereas robotic surgery with fluorescent ureteral catheters was used in selected patients. No intraoperative ureteral injury was observed in the stent group. Catheter-related complications, including hematuria (14.3%) and urinary tract infections (9.5%), were minor and resolved before discharge. Conclusions: PUC may be beneficial in patients with a history of pelvic surgery or local recurrence of rectal cancer, in whom the risk of ureteral injury is inherently higher. Full article

Background: To enhance the bioavailability and neuroprotective efficacy of biatractylolide against Alzheimer’s disease by developing a novel Tween-80-modified pullulan–chenodeoxycholic acid nanoparticle as a delivery vehicle. Methods: Chenodeoxycholic acid (CDCA) was chemically conjugated to pullulan to yield hydrophobically modified pullulan (PUC), onto which polysorbate 80 (Tween-80) was subsequently adsorbed. The PUC polymers with CDCA substitution levels were analyzed by ¹H NMR spectroscopy. Nanoparticles were fabricated via the dialysis method and characterized by transmission electron microscopy and dynamic light scattering for morphology, size, and surface charge. In vitro neuroprotection was assessed by exposing SH-SY5Y and PC12 cells to 20 µM Aβ_25-35 to induce cytotoxicity, followed by pretreatment with biatractylolide-loaded PUC (BD-PUC) nanoparticle solutions at various biatractylolide concentrations. The in vivo brain-targeting capability of both empty PUC and BD-PUC particles was evaluated using a live imaging system. Results: The ¹H NMR analysis confirmed three distinct CDCA substitution degrees (8.97%, 10.66%, 13.92%). Transmission electron microscopy revealed uniformly dispersed, spherical nanoparticles. Dynamic light scattering measurements showed a hydrodynamic diameter of ~200 nm and a negative zeta potential. Exposure to 20 µM Aβ_25-35 significantly reduced SH-SY5Y and PC12 cell viability; pretreatment with BD-PUC nanoparticles markedly enhanced cell survival rates and preserved cellular morphology compared to cells treated with free biatractylolide. Notably, the cytoprotective effect of BD-PUC exceeded that of the free drug. In vivo imaging demonstrated that both empty PUC and Tween-80-adsorbed BD-PUC nanoparticles effectively accumulated in the brain. Conclusions: The protective effect of BD-PUC on SH-SY5Y and PC12 cells induced by Aβ_25-35 was higher than free biatractylolide solution, and the BD-PUC nanosolution modified with Tween-80 showed a brain-targeting effect. Full article

Intersection sight distance (ISD) requirements, currently designed for driver-operated vehicles (DVs), will be affected once automated vehicles (AVs) enter the driving environment. This paper examines the ISD for intersections with a yield control on a minor road in a mixed DV-AV environment. Five potential conflict types with different ISD requirements are modeled as a minor-road vehicle proceeds to cross the intersection, turns right, or turns left. Furthermore, different models are developed for each conflict type depending on the vehicle types on the minor and major roads. These models, along with the intersection geometry, establish the system demand and supply models for ISD reliability analysis. A surrogate safety measure is developed and used to measure ISD non-compliance and is denoted by the probability of unresolved conflicts (PUC). The models are applied to a case study intersection, where PUC values are estimated using Monte Carlo Simulation and compared to an established target value relating to the DV-only traffic of 0.00674. The results show that AV-related traffic has higher overall PUC values than those of DV-only traffic. A corrective measure, reducing the AV speed limit on the minor-road approaches by 3 to 4 km/h, decreases the overall PUC to values below those of the target PUC. Full article

Polyurethane concrete (PUC) is a promising candidate for structural repair materials due to its excellent mechanical properties and durability. However, the bonding performance between PUC and concrete interfaces may limit its broader application. This study examined the factors affecting the shear strength at the PUC–NC interface. A total of 16 oblique shear tests, varying by interface treatment methods (smooth—GH, roughened—ZM, and grooved—KC), adhesive application rates—NJJ (0, 0.2, and 0.3 kg/m²), and steel fiber contents—GXW (0%, 0.5%, 1%, and 1.5%), to evaluate their impact on the mechanical properties of the PUC–NC interface. The results demonstrated that roughening the interface significantly improved the shear strength, resulting in a 32% increase compared to a smooth interface and 15% compared to a grooved interface. A moderate adhesive application rate (0.2 kg/m²) enhanced the interface strength, while excessive adhesive did not further increase the shear strength. The optimal steel fiber content (1%) resulted in the highest shear strength, improving it by 22%, whereas excess steel fibers (1.5%) reduced the interface strength. This is due to fiber agglomeration, which weakens mechanical interlocking and introduces defects that impair interfacial bonding. Load–slip curve analysis revealed that roughened interfaces combined with the appropriate amount of steel fibers improved the interface toughness, delaying the failure process. This study presents a model for calculating the shear strength of steel fiber-reinforced PUC–NC interfaces, incorporating shear slip. Compared to existing models, it more accurately reflects the experimental data. Full article

The concept of positive mental health (PMH) and therefore mental well-being (MWB) have taken center stage over the last few decades. The Mental Health Continuum—Short Form (MHC-SF) is one of the most important tools for assessing MWB. This study aims to assess the psychometric properties and dimensionality of the Portuguese version of the MHC-SF by comparing three competing models. Methods: Between January and March of 2021, a survey was conducted with a convenience sample of 495 Portuguese adolescents aged 12 to 18 years, enrolled in grades 7 to 12. Descriptive statistics and bivariate statistical tests and measures associated with confirmatory factor analysis (CFA) were calculated. Results: The results of the CFA support the idea that the bifactor model fits the data better than the other competing models. The indices support unidimensionality, namely the explained common variance (ECV = 0.81), the Percentage of Uncontaminated Correlations (PUC = 0.69), and the omega hierarchical (ωH = 0.89), and point to the existence of a general MHC-SF factor. The scale showed high reliability (ω = 0.95) in the general factor. The MHC-SF has concurrent validity, correlating with other scales that assess aspects related to mental health and well-being. Conclusion: The results point to the adequacy of the bifactor model and suggest the existence of a general factor of PMH, confirming that the total score of the scale can be used. Full article

Metal complexes have potential applications in drug discovery due to their unique properties. For example, zinc(II) ions (Zn^II) exhibit a high affinity for DNA and have been used as active centers in artificial protein/small-molecule metallonucleases. In this study, we designed a series of ligands containing a biaryl moiety as a photosensitizer to synthesize Zn^II complexes with enhanced DNA affinity for use as DNA photocleavage reagents. The DNA photocleavage activity of these complexes was evaluated using the pUC19 plasmid, revealing that the Zn^II complex bearing the 4′-biphenyl-bpa (bpa = bis(2-picolyl)amine) ligand L1 exhibited the strongest DNA photocleavage activity. Further analysis of the biological activity of the Zn^II complex of L1 in the human pancreatic cancer cell line MIA PaCa-II demonstrated that its cytotoxic activity increased in a UV irradiation time-dependent manner, with an IC₅₀ value of 14.2 μM. Fluorescence staining revealed that the Zn^II complex of L1 generates reactive oxygen species in cells, leading to DNA double-strand breaks upon UV irradiation and ultimately resulting in necrotic cell death. These findings highlight the potential of the Zn^II complex of L1 as a photochemotherapeutic agent for pancreatic cancer. Full article

In this research, the structure, morphology, hydrophobicity and swelling behavior of a polyurethane (PU) network and its composites (PUCs) were examined. PUCs were synthesized by the incorporation of different percentages (0.5, 1 and 2 wt.%) of unmodified or surface-modified TiO₂ nanoparticles into a PU network based on polycaprolactone, aliphatic hyperbranched polyester and isophorone diisocyanate. In order to improve interfacial interactions, the surface of the TiO₂ nanoparticles was chemically modified with lauryl gallate. The impact of the presence and content of unmodified or surface-modified TiO₂ nanoparticles on the cross-sectional and surface morphology, swelling behavior and hydrophobicity of the PU network was assessed by different experiments. The obtained findings revealed that the incorporation of TiO₂ nanoparticles brought a more pronounced irregular cross-sectional and rougher surface morphology, better microphase separation, higher values of the equilibrium swelling degree in tetrahydrofuran and toluene, and altered water contact angles compared to the neat PU. Based on the collected results, the practical applicability of the prepared PUCs may be in the area of protective coatings. Full article

To meet the increasing demand for resilient infrastructure in seismic and high-impact areas, accurate prediction and reliability analysis of the performance of composite structures under impact loads is essential. Conventional techniques, including experimental testing and high-quality finite element simulation, require considerable time and resources. To address these issues, this study investigated individual and hybrid models including support vector regression (SVR), Gaussian process regression (GPR), and improved eliminate particle swamp optimization hybridized artificial neural network (IEPANN) models for predicting the failure strength of composite concrete developed by combining normal concrete (NC) with ultra-high performance concrete (UHPC) and polyurethane-based polymer concrete (PUC), considering different surface treatments and subjected to various static and impact loads. An experimental dataset was utilized to train the ML models and perform the reliability analysis on the impact dataset. Key parameters included compressive strength (Cfc), flexural load of the U-shaped specimens (P), density (ρ), first crack strength (N1), and splitting tensile strength (ft). Results revealed that all the developed models had high prediction accuracy, achieving NSE values above acceptable thresholds greater than 90% across all the datasets. Statistical errors such as RMSE, MAE, and PBIAS were calculated to fall within acceptable limits. Hybrid IEPANN appeared to be the most effective model, demonstrating the highest NSE value of 0.999 and the lowest RMSE, PBIAS, and MAE values of 0.0013, 0.0018, and 0.001, respectively. The reliability analysis revealed that impact times (N1 and N2) reduced as the survival probability increased. Full article

Multilevel inverters have gained importance in modern power systems during the last few years because of their high power quality with lower THD. Various topologies developed include the packed U-cell inverter and its different modified versions that have emerged as a compact and efficient solution to distributed energy systems. Most of the available harmonic mitigation techniques, that is, passive filtering and individual optimization techniques, which include GA and PSO, are susceptible to a variety of shortcomings regarding their inherent complexity and inefficiency; hence, finding an appropriate convergence may be quite hard. This paper proposes a hybrid version of the GA-PSO algorithm that exploits the exploratory strengths of GA and the convergence efficiencies of PSO in determining the optimized switching angles for SHM techniques applied to modified five-level and seven-level PUC inverters. By utilizing the multi-objective optimization method, the approach minimizes THD while keeping voltage and efficiency constraints. Simulated in MATLAB/Simulink, the results were experimentally verified using hardware-in-the-loop testing on OP5700. A large THD reduction in both MPUC7 (11.68%) and MPUC5 (17.61%) was obtained. The proposed hybrid algorithm outperformed the standalone approaches of GA and PSO with respect to robustness and with precise harmonic suppression. Other appealing features are reduced computational complexity and improved waveform quality; hence, the method is highly suitable for both grid-tied and standalone renewable energy applications. This work lays a basis for efficient inverter designs that can adapt well under dynamic load conditions. Full article

To clone DNA sequences quickly and precisely into plasmids is essential for molecular biology studies. Some cloning vectors have been developed for the cloning of PCR products, including blunt-end and T-A cloning. However, different plasmids are required for the cloning of PCR products with blunt ends and 3′ A overhang ends. Here, a novel cloning vector, pYFRed, which is based on the pUC19 backbone, has emerged and can be applied in both blunt-end and T-A cloning. PCR products can be cloned into the pYFRed by a one-step digestion–ligation reaction in a tube. The endonuclease recognition sequences of SmaI, Eco53kI, EcoRV, PmeI, and SwaI for blunt-end cloning and XcmI for T-A cloning were designed and added between the lac promoter and the starting codon ATG of the mScarlet-I gene of pYFRed. The ligation efficiency was significantly higher because the restriction enzyme sites utilized were removed from the vector after being successfully constructed. The mScarlet-I gene was introduced into the pYFRed for the screening of the positive recombinants by the unaided eye without the need for additional reagents/equipment. pYFRed is easy to construct in an ordinary laboratory, which facilitates researchers to develop their cloning vector without purchasing commercial cloning vectors. Full article

Background/Objectives: Data on the real-world effectiveness and safety of selective JAK inhibitors (JAKis) in ulcerative colitis (UC) and Crohn’s disease (CD) are limited. Methods: We conducted a multicentre, retrospective study to assess clinical, biochemical, and endoscopic outcomes of selective JAKis in bio-experienced UC and CD. Results: A total of 246 patients (mean age: 40.5 ± 14.5 years; 131 UC and 115 CD) were included with a median follow-up of 7.5 months. Among the CD patients receiving upadacitinib (n = 115), 76.2% achieved clinical remission (CR) at week 12. Furthermore, 59.5% of the upadacitinib-treated UC patients (n = 100) experienced CR at week 8. Corticosteroid-free CR (CSFCR) was achieved by 76.9% of the CD patients and 80.6% of the UC patients at week 24, while 50.0% and 36.1% experienced endoscopic remission. At week 52, 66.7% of the CD and 86.2% of the UC patients achieved CSFCR, whereas 54.5% and 52.9% had endoscopic remission. In UC, the effectiveness of upadacitinib was not compromised by prior tofacitinib failure, while the upadacitinib-treated CD patients with stricturing and penetrating disease were less likely to achieve CR by the end of the induction phase (p = 0.04). C-reactive protein (p[CD] < 0.0001; p[UC] < 0.0001) and faecal calprotectin (p[CD] < 0.0001; p[UC] = 0.02) decreased significantly in both patient groups as early as week 2. Among the filgotinib-treated UC patients (n = 31), 28.6% were in CR at week 12. At week 24 and 52, 59.1% and 60% achieved CSFCR, while 0.0% and 20.0% had endoscopic remission. Both C-reactive protein (p = 0.04) and faecal calprotectin (p = 0.04) decreased significantly by week 12. Hyperlipidaemia (9.7–9.8%) was the most common adverse event. Conclusions: Selective JAKis are rapidly effective and safe for treating refractory, moderate-to-severe CD and UC. Full article

This study examines the critical role of inclusive, people-centered strategies in driving organizational sustainability, focusing on two key institutions in the Ashanti Region of Ghana: Presbyterian University College (PUC) and Presbyterian Agogo Women’s College of Education (APWCE). Employing a qualitative research design, including 100 interviews, five focus groups, and participant observations, this study investigates employee perspectives on Diversity, Equity, and Inclusion (DEI) practices in the workplace. The findings identify key empowerment strategies—flexible job roles, participatory decision-making, leadership development, and open communication—that enhance employee engagement and commitment to sustainability efforts. The findings also demonstrate employees’ vital role in advancing sustainability through involvement in green initiatives, community engagement, and integrating sustainability into core organizational practices. This contribution intellectually bridges the gap between DEI policies and their practical application, offering a nuanced understanding of how cultural and social dimensions influence sustainability in underexplored contexts like Ghana. It emphasizes aligning organizational values with employee well-being to enhance job satisfaction and retention, presenting actionable strategies for fostering innovation, resilience, and long-term success. The increasing global focus on sustainability and the growing need for inclusive practices in organizational settings underscores the timeliness of this manuscript. It offers a holistic, forward-thinking approach that is especially relevant for organizations navigating post-pandemic workplace dynamics and seeking to align sustainability with equity and inclusivity. Full article

The antibiotic resistance crisis has seriously jeopardized public health and human safety. As one of the ways of horizontal transfer, transformation enables bacteria to acquire exogenous genes naturally. Bisphenol compounds are now widely used in plastics, food, and beverage packaging, and have become a new environmental pollutant. However, their potential relationship with the spread of antibiotic resistance genes (ARGs) in the environment remains largely unexplored. In this study, we aimed to assess whether the ubiquitous bisphenol S (BPS) could promote the transformation of plasmid-borne ARGs. Using plasmid pUC19 carrying the ampicillin resistance gene as an extracellular ARG and model microorganism E. coli DH5α as the recipient, we established a transformation system. Transformation assays revealed that environmentally relevant concentrations of BPS (0.1–10 μg/mL) markedly enhanced the transformation frequency of plasmid-borne ARGs into E. coli DH5α up to 2.02-fold. Fluorescent probes and transcript-level analyses suggest that BPS stimulated increased reactive oxygen species (ROS) production, activated the SOS response, induced membrane damage, and increased membrane fluidity, which weakened the barrier for plasmid transfer, allowing foreign DNA to be more easily absorbed. Moreover, BPS stimulates ATP supply by activating the tricarboxylic acid (TCA) cycle, which promotes flagellar motility and expands the search for foreign DNA. Overall, these findings provide important insight into the role of bisphenol compounds in facilitating the horizontal spread of ARGs and emphasize the need to monitor the residues of these environmental contaminants. Full article