Managing cellular networks in the Beyond 5G (B5G) era is a complex and challenging task requiring advanced deep learning approaches. Traditional models focusing on internet traffic (INT) analysis often fail to capture the rich temporal and spatial contexts essential for accurate INT predictions. Furthermore, these models do not account for the influence of external factors such as weather, news, and social trends. This study proposes a multi-source CNN-RNN (MSCR) model that leverages a rich dataset, including periodic, weather, news, and social data to address these limitations. This model enables the capture and fusion of diverse data sources for improved INT prediction accuracy. An advanced deep learning model, the transformer-enhanced CNN-RNN (TE-CNN-RNN), has been introduced. This model is specifically designed to predict INT data only. This model demonstrates the effectiveness of transformers in extracting detailed temporal-spatial features, outperforming conventional CNN-RNN models. The experimental results demonstrate that the proposed MSCR and TE-CNN-RNN models outperform existing state-of-the-art models for traffic forecasting. These findings underscore the transformative power of transformers for capturing intricate temporal-spatial features and the importance of multi-source data and deep learning techniques for optimizing cell site management in the B5G era. Full article

Air-to-air heat pumps are used in today’s ventilation systems increasingly often as they provide heating and cooling for buildings. The energy transformation modes of these units are subject to constant change due to the varying outdoor air state, including temperature and humidity. When choosing how to operate and control energy transformers, it is important to be able to adapt effectively to the changing outside air conditions. Nowadays, modern commercial heat pumps offer two levels of control flexibility: a compressor with a variable speed and an electronic expansion valve. This combination of control elements has boosted the seasonal energy efficiency of heat pumps. For a long time, cycle control possibilities have been dominated by electronic controls. The authors of this paper aim to present an additional element to the traditional heat pump controls, which provides a third level of control over the cycle. To achieve the objective, experimental investigations of a heat pump integrated into a ventilation unit have been carried out under real-life conditions. The experiments involved varying the operating modes of the unit by adjusting the compressor speed, the position of the expansion valve, and the volume of the system loop. The study examined the performance characteristics of the heat pump and found that the performance of a variable-volume heat pump is comparable to that of a conventionally operated typical constant-volume heat pump system. In addition, the study found that by adding a third level of volume control to the active heating circuit, in combination with conventional controls, the heat pump’s heat output range could be extended by 69.62%. The study determined the variation of the heat pump cycle in the p-h diagram with the variation of the loop volume. The benefits and drawbacks of a heat pump with a variable-volume loop are discussed in this study. Full article

The nematicidal potential of the yeast Papiliotrema terrestris strain PT22AV (YSY) was investigated against the root nematode (RKN) Meloidogyne incognita in in vitro bioassays on infective juveniles (J2) and experiments on tomatoes in pot and greenhouse conditions. The J2 nematodes were exposed to YSY solutions for 19 days, using abamectin (ABA), fosthiazate (FOS) and distilled water as controls. In the experiments on potted and greenhouse tomatoes, 0.5 and 1 kg ha⁻¹ doses of YSY were tested in comparison to ABA, biocontrol agents Purpureocillium lilacinus strain 251 (PUL) and Bacillus firmus strain 1-1582 (BAF), a plant biostimulant/fertilizer (ERG) and the nematicide Fluopyram (FLU). J2’s viability was affected by YSL after 7 days, decreasing to zero on the 15th exposure day, while ABA and FOS resulted in 83 and 100% J2 mortality within 24 h. Only the 1.0 kg ha⁻¹ dose of YSY was able to significantly reduce the final nematode population in soil and gall formation on tomato roots, without significant differences from PUL and BAF. All treatments in comparison also resulted in a significant increase in tomato growth and crop yield, except for 0.5 kg ha⁻¹ of YSY. Data indicated that YSY could represent an additional tool for organic and integrated RKN management. Full article

This study introduces a new machine learning-based algorithm for the retrieving significant wave height (SWH) using synthetic aperture radar (SAR) images. This algorithm is based on the azimuthal cut-off wavelength and was developed in quad-polarized stripmap (QPS) mode in coastal waters. The collected images are collocated with a wave simulation from the numeric model, called WAVEWATCH-III (WW3), and the current speed from the HYbrid Coordinate Ocean Model (HYCOM). The sea surface wind is retrieved from the image at the vertical–vertical polarization channel, using the geophysical model function (GMF) CSARMOD-GF. The results of the algorithm were validated against the measurements obtained from the Haiyang-2B (HY-2B) scatterometer, yielding a root mean squared error (RMSE) of 1.99 m/s with a 0.82 correlation (COR) and 0.27 scatter index of wind speed. It was found that the SWH depends on the wind speed and azimuthal cut-off wavelength. However, the current speed has less of an influence on azimuthal cut-off wavelength. Following this rationale, four widely known machine learning methods were employed that take the SAR-derived azimuthal cut-off wavelength, wind speed, and radar incidence angle as inputs and then output the SWH. The validation result shows that the SAR-derived SWH by eXtreme Gradient Boosting (XGBoost) against the HY-2B altimeter products has a 0.34 m RMSE with a 0.97 COR and a 0.07 bias, which is better than the results obtained using an existing algorithm (i.e., a 1.10 m RMSE with a 0.77 COR and a 0.44 bias) and the other three machine learning methods (i.e., >a 0.58 m RMSE with a <0.95 COR), i.e., convolutional neural networks (CNNs), Support Vector Regression (SVR) and the ridge regression model (RR). As a result, XGBoost is a highly efficient approach for GF-3 wave retrieval at the regular sea state. Full article

The purpose of this study was to evaluate the quality of recovery from general anesthesia with the administration of two low doses of dexmedetomidine in canine patients. For this blind randomized clinical trial study, 30 dogs undergoing general anesthesia for diagnostic procedures or elective surgery (ovariectomy/castration) were included. The patients were randomly divided into three groups, and at the end of anesthesia, they received a bolus of dexmedetomidine at 1 mcg/kg IV (D1), or a bolus of dexmedetomidine at 0.5 mcg/kg (D0.5), or a bolus of NaCl, in a total of 0.5 mL of solution for all three groups. After administration of the bolus, the anesthetist monitored the patients every 5 min by measuring heart rate, systolic and mean blood pressure, respiratory rate, and oxygen saturation. The quality of recovery was also assessed using 4 different scales. The extubation time, time of headlift, and standing position were also recorded. Both groups receiving dexmedetomidine had better awakening and a lower incidence of delirium when compared to saline administration. The heart rate was lower, while the systolic pressure was higher in the two groups D1 and D0.5 compared to the NaCl with a low presence of atrioventricular blocks. The extubation time resulted significantly higher in the D1 (17 ± 6 min) compared to the D0.5 (10 ± 4 min) and NaCl (8 ± 3 min) (p < 0.0001); the headlift time D1 (25 ± 10 min) resulted significantly longer than the NaCl group (11 ± 5 min) (p = 0.0023) but not than the D0.5 (18 ± 9 min). No significant differences were found among the three groups for standing positioning (D1 50 ± 18 min, D0.5 39 ± 22 min, NaCl 28 ± 17 min). The preventive administration of a bolus of dexmedetomidine at a dosage of 0.5 mcg/kg or 1 mcg/kg IV during the recovery phase improves the quality of recovery in patients undergoing general anesthesia. Full article

In recent years, the digital economy has shown great potential in regard to in driving social production and development. In the context of the construction of digital villages, the deep integration of the digital economy and agricultural development has injected new vitality into improving the quality and efficiency of agricultural production, becoming an important way to promote sustainable agricultural development. Based on the panel data of 31 provinces in China from 2012 to 2021, the study utilizes the entropy method to measure the level of the digital economy and the high-quality development of agriculture. Additionally, this study explores the impact and mechanism of the digital economy on the high-quality development of agriculture by the fixed effect, mediation effect, and the spatial spillover models. In summary, the digital economy can significantly drive the high-quality development of agriculture, which is still valid after considering endogeneity and robustness. Mechanistically, the rationalization of industrial structure is an important path for the digital economy in regard to driving the high-quality development of agriculture. Regionally, the dividends of the digital economy for high-quality agricultural development in the central and western regions are greater than those in the eastern region. Spatially, the digital economy has a spatial spillover effect on the high-quality development of agriculture. Moreover, it can promote the synergistic development of adjoining regions. Therefore, policy recommendations are made in terms of strengthening rural infrastructure, emphasizing the development of regional shortcomings, and strengthening internal with external regional linkages. Full article

Flood hazards present a significant risk to the UK, with homes, businesses and critical infrastructure exposed to a mixture of fluvial, surface water and coastal flooding. Climate change is expected to influence river flows, changing the frequency and magnitude of future flood events. Flood hazard assessments are used by decision-makers to implement policies and engineering interventions to reduce the impacts of these flood events. Probabilistic flood modelling can explore input and parameter uncertainties in flood models to fully quantify inundation uncertainty. However, probabilistic methods require large computational costs—limiting their application. This paper investigates a range of advanced uncertainty quantification methods (traditional Monte Carlo (FMC), Kriging and multi-fidelity Monte Carlo (MFMC)) to reduce the dichotomy between accuracy and costs. Results suggest that Kriging can reduce computational costs by 99.9% over FMC. The significantly increased efficiency has the potential to improve future policy and engineering decisions, reducing the impacts of future flood events. Full article

Background and Objectives: Current guidelines and the alteplase product insert recommend that antithrombotic therapy be avoided within 24 h of intravenous thrombolytic therapy with rt-PA in acute ischemic stroke. Therefore, the rate of stroke recurrence is unclear in terms of early neurological deterioration, which we could prevent with the early administration of antithrombotic therapy. We do not know the effect of early antithrombotic therapy after intravenous thrombolysis with rt-PA in acute stroke on the outcome in patients after 90 days either. Design: Prospective monocentric observational cohort study. Methods: Data were collected from consecutive patients treated with alteplase for acute ischemic stroke between January 2015 and January 2023. We examined functional outcome at 90 days, including the risk of symptomatic intracranial hemorrhage and mortality rate as safety indicators and stroke recurrence events in both early and standard antithrombotic therapy at 24 h after intravenous thrombolysis. Results: A total of 489 patients were included, of which 278 (56.9%) were men. Of these, 407 (83.2%) patients received early antithrombotic therapy. No symptomatic intracranial hemorrhage occurred in any participants. There was a significantly higher number of patients with an excellent outcome (mRS 0-1) in early antithrombotic treatment (211 (53.1%) versus 28 (34.6%) in standard antithrombotic treatment (p = 0.002, OR 0.47, 95% CI: 0.28–0.76). Conclusions: Early antithrombotic treatment after intravenous therapy in patients with acute ischemic stroke revealed no safety concerns compared with standard antithrombotic therapy and resulted in a significantly higher proportion of patients with an excellent functional outcome. Full article

Piceatannol (PIC), a polyphenol abundant in passion fruit seeds, is reported to promote fat metabolism. This study investigated whether PIC affects sirtuin 1 (SIRT1) expression and metabolic factors in C2C12 skeletal muscle cells. C2C12 myotubes were stimulated with PIC, and alterations in gene expression, protein levels, mitochondrial DNA content, and fatty acid levels were assessed using real-time PCR, Western blotting, and Nile red staining. Furthermore, we examined changes in SIRT1 expression following the consumption of a test food containing 100 mg PIC for 2 weeks among adults with varying age and body mass index ranges. Both PIC and passion fruit seed extract induced SIRT1 expression in C2C12 myotubes to a greater extent than resveratrol. PIC also increased the expression of genes associated with mitochondrial biogenesis and fatty acid utilization, increased mitochondrial DNA content, and suppressed oleic acid-induced fat accumulation. Moreover, participants who consumed PIC exhibited significantly higher SIRT1 mRNA expression in whole blood compared to those in the placebo group. These findings suggest that PIC induces SIRT1 expression both in vitro and in the human body, which may promote mitochondrial biosynthesis and fat metabolism. Full article

Eight balls were manufactured with a 3D printer to resemble various types of 32-panel soccer balls. One ball was completely smooth, whereas the other seven possessed various dimple patterns on their surface panels. Seam width and seam depth were also varied. Wind-tunnel experiments were performed to extract aerodynamic coefficients, and also to determine the critical Reynolds number for each manufactured ball. A new surface roughness parameter is introduced, and a fitting formula is presented, which allows for the prediction of the critical Reynolds number if the new parameter is known. Full article

The azimuth track is an important component of the radio telescope wheel–rail system. During operation, the azimuth track is inevitably subject to phenomena such as track wear, track fatigue cracks, and impact damage to welded joints, which can affect observation accuracy. The 110 m QiTai radio telescope (QTT) studied in this paper is the world’s largest fully steerable radio telescope at present, and its track will bear the largest load ever. Since the welded joint of an azimuth track is the weakest part, an innovative welding method (multi-layer and multi-pass weld) is adopted for the thick welding section. Therefore, it is necessary to study the contact mechanical properties between the wheel and the azimuth track in this welded joint. In this study, tensile tests based on digital image correlation technology (DIC) and Vickers hardness tests are carried out in the metal zone (BM), heat-affected zone (HAZ), modified layer, and weld zone (WZ) of the welded joint, and the measured data are used to fit the elastic–plastic constitutive model for the different zones of the welded joint in the azimuth track. Based on the constitutive model established, a nonlinear finite element model is built and used to simulate the rolling mechanical performance between the wheel and azimuth track. Through the analysis of simulated data, we obtained the stress distribution of the track under different pre-designed loads and identified the locations most susceptible to damage during ordinary working conditions, braking conditions, and start-up conditions. The result can provide a significant theoretical basis for future research and for the monitoring of large track damage. Full article

As dengue expands globally and many vaccines are under trials, there is a growing recognition of the need for assessing T cell immunity in addition to assessing the functions of neutralizing antibodies during these endeavors. While several dengue-specific experimentally validated T cell epitopes are known, less is understood about which of these epitopes are conserved among circulating dengue viruses and also shared by potential vaccine candidates. As India emerges as the epicenter of the dengue disease burden and vaccine trials commence in this region, we have here aligned known dengue specific T cell epitopes, reported from other parts of the world with published polyprotein sequences of 107 dengue virus isolates available from India. Of the 1305 CD4 and 584 CD8 epitopes, we found that 24% and 41%, respectively, were conserved universally, whereas 27% and 13% were absent in any viral isolates. With these data, we catalogued epitopes conserved in circulating dengue viruses from India and matched them with each of the six vaccine candidates under consideration (TV003, TDEN, DPIV, CYD-TDV, DENVax and TVDV). Similar analyses with viruses from Thailand, Brazil and Mexico revealed regional overlaps and variations in these patterns. Thus, our study provides detailed and nuanced insights into regional variation that should be considered for itemization of T cell responses during dengue natural infection and vaccine design, testing and evaluation. Full article

Oceanic energy sources, notably offshore wind and wave power, present a significant opportunity to generate green hydrogen through water electrolysis. This approach allows for offshore hydrogen production, which can be efficiently transported through existing pipelines and stored in various forms, offering a versatile solution to tackle the intermittency of renewable energy sources and potentially revolutionize the entire electrical grid infrastructure. This research focusses on assessing the technical and economic feasibility of this method in six strategic coastal regions in Morocco: Laayoune, Agadir, Essaouira, Eljadida, Casablanca and Larache. Our proposed system integrates offshore wind turbines, oscillating water column wave energy converters, and PEM electrolyzers, to meet energy demands while aligning with global sustainability objectives. Significant electricity production estimates are observed across these regions, ranging from 14 MW to 20 MW. Additionally, encouraging annual estimates of hydrogen production, varying between 20 and 40 tonnes for specific locations, showcase the potential of this approach. The system’s performance demonstrates promising efficiency rates, ranging from 13% to 18%, while maintaining competitive production costs. These findings underscore the ability of oceanic energy-driven green hydrogen to diversify Morocco’s energy portfolio, bolster water resilience, and foster sustainable development. Ultimately, this research lays the groundwork for comprehensive energy policies and substantial infrastructure investments, positioning Morocco on a trajectory towards a decarbonized future powered by innovative and clean technologies. Full article

The biological degradation of plant residues in the soil or on the soil surface is an integral part of the natural life cycle of annual plants and does not have adverse effects on the environment. Crop straw is characterized by a complex structure and exhibits stability and resistance to rapid microbial decomposition. In this study, we conducted a microcosm experiment to investigate the dynamic succession of the soil microbial community and the functional characteristics associated with lignocellulose-degrading pathways. Additionally, we aimed to identify lignocellulose-degrading microorganisms from the straw of three crop species prevalent in Northeast China: soybean (Glycine max Merr.), rice (Oryza sativa L.), and maize (Zea mays L.). Our findings revealed that both the type of straw and the degradation time influenced the bacterial and fungal community structure and composition. Metagenome sequencing results demonstrated that during degradation, different straw types assembled carbohydrate-active enzymes (CAZymes) and KEGG pathways in distinct manners, contributing to lignocellulose and hemicellulose degradation. Furthermore, isolation of lignocellulose-degrading microbes yielded 59 bacterial and 14 fungal strains contributing to straw degradation, with fungi generally exhibiting superior lignocellulose-degrading enzyme production compared to bacteria. Experiments were conducted to assess the potential synergistic effects of synthetic microbial communities (SynComs) comprising both fungi and bacteria. These SynComs resulted in a straw weight loss of 42% at 15 days post-inoculation, representing a 22% increase compared to conditions without any SynComs. In summary, our study provides novel ecological insights into crop straw degradation by microbes. Full article

Coronaviruses represent a significant class of viruses that affect both animals and humans. Their replication cycle is strongly associated with the endoplasmic reticulum (ER), which, upon virus invasion, triggers ER stress responses. The activation of the unfolded protein response (UPR) within infected cells is performed from three transmembrane receptors, IRE1, PERK, and ATF6, and results in a reduction in protein production, a boost in the ER’s ability to fold proteins properly, and the initiation of ER-associated degradation (ERAD) to remove misfolded or unfolded proteins. However, in cases of prolonged and severe ER stress, the UPR can also instigate apoptotic cell death and inflammation. Herein, we discuss the ER-triggered host responses after coronavirus infection, as well as the pharmaceutical targeting of the UPR as a potential antiviral strategy. Full article

This article explores the scientific and practical challenges associated with developing simulation modeling methods for remanufacturing within a circular economy framework. It aims to define this concept and identify the key hurdles that need to be overcome for its successful implementation. According to the six principles of sustainable development, the key pillar is remanufacturing. Remanufacturing helps the environment in several different ways, including: saving energy, the conservation of raw materials, the conservation of space, landfills, the reduction of air pollution, and greater fuel efficiency. This process closes the loop in the supply chain, exemplifying the principles of a circular economy. The research methods used are primarily the analysis and criticism of literature, document examination—especially in relation to existing simulation programs and analysis—the logical construction method, and the heuristic method, used to define concept of simulation modelling. In response to scientific and practical challenges, the concept of a new modeling method was defined and presented. This concept uses the legacy of Lean and the author’s original ideas regarding the structuring of the remanufacturing factory and processes. The main contribution of this study is integration, embedding this concept into the simulation software. A comparison with existing solutions and the advantages of the new concept are also included in the article. Full article

Background: Calprotectin (CP) is a calcium- and zinc-binding protein that plays a key role in innate immunity and in the recruitment of inflammatory cells. CP can be detected both in serum and in fecal samples. Serum CP (sCP) is more specific for autoimmune diseases, while fecal CP (fCP) has been well investigated for gastrointestinal diseases. Few studies have shown the clinical effectiveness of sCP as an acute-phase biomarker for gastrointestinal diseases. Aim: The aim of this narrative review is to discuss the role of sCP as a useful alternative biomarker of the acute-phase activity of gastrointestinal diseases and as a possible tool for screening and monitoring these diseases. Material and Methods: We searched original articles, abstracts, reviews, case reports, and clinical trials on PubMed^®, Up-to-Date^®, and Medscape^® in the last ten years. Conclusion: We found that sCP could represent a useful biomarker in the evaluation of the inflammatory stage in patients with immune-mediated gastrointestinal diseases, but more studies are needed to promote its routine use in clinical practice as a diagnostic and prognostic biomarker as a replacement for fCP. Full article

Systemic sclerosis (SSc) is a rare autoimmune connective tissue disorder characterized by massive fibrosis, vascular damage, and immune imbalance. Advances in rheumatology and immunology over the past two decades have led to a redefinition of systemic sclerosis, shifting from its initial perception as primarily a “hyperfibrotic” state towards a recognition of systemic sclerosis as an immune-mediated disease. Consequently, the search for genetic markers has transitioned from focusing on fibrotic mechanisms to exploring immune regulatory pathways. Immunogenetics, an emerging field at the intersection of immunology, molecular biology, and genetics has provided valuable insights into inherited factors that influence immunity. Data from genetic studies conducted thus far indicate that alterations in genetic messages can significantly impact disease risk and progression. While certain genetic variations may confer protective effects, others may exacerbate disease susceptibility. This paper presents a comprehensive review of the most relevant genetic changes that influence both the risk and course of systemic sclerosis. Special emphasis is placed on factors regulating the immune response, recognizing their pivotal role in the pathogenesis of the disease. Full article

Zirconium carbide (ZrC) ceramics have a high melting point, low neutron absorption cross section, and excellent resistance to the impact of fission products and are considered to be one of the best candidate materials for fourth-generation nuclear energy systems. ZrC ceramics with a high relative density of 99.1% were successfully prepared via pressureless sintering using a small amount of MoSi₂ as an additive. The influence of the MoSi₂ content on the densification behavior, microstructure, mechanical properties, and thermal properties of ZrC ceramics was systematically investigated. The results show that the densification of ZrC was significantly enhanced by the introduction of MoSi₂ due to the formation of a liquid phase during sintering. In addition, the ZrC grains were refined due to the pinning effect of the generated silicon carbide. The flexural strength and Vickers hardness of ZrC ceramics with 2.5 vol% MoSi₂ sintered at 1850 °C were 408 ± 12 MPa and 17.1 GPa, respectively, which were approximately 30% and 10% higher compared to the samples without the addition of MoSi₂. The improved mechanical properties were mainly attributed to the high relative density (99.1%) and refined microstructure. Full article

4-nitrophenol (4-NP) is a frequently encountered toxic phenolic organic pollutant in water. It is important to develop a simple method to treat 4-NP. Small and monodispersed gold nanoparticles often have good catalytic performance of 4-NP. Hemoglobin (Hb) is a kind of common and important protein in organisms. Herein, highly biocompatible bovine hemoglobin-stabilized gold nanoparticles (Au_n-Hb NPs) were synthesized using hemoglobin as a biological template. Then, the size, zeta potential, and composition of Au_n-Hb NPs were investigated by transmission electron microscopy, dynamic light scattering, and X-ray photoelectron spectroscopy. The Au_n-Hb NPs with small gold nanoparticles of about 1.4–2.4 nm had good catalytic capabilities in reducing 4-NP to form 4-aminophenol. Au₂₀-Hb NPs demonstrated superior catalytic efficiency in the reduction of 4-NP when compared to other nanoparticles. Moreover, as-synthesized Au₂₀-Hb NPs exhibited excellent biocompatibility through the MTT experiment. The method of preparation of gold nanoparticles offers one way to prepare metal nanoparticles for good potential catalytic applications of gold nanoparticles. Full article

This work investigated material extrusion additive manufacturing (MatEx AM) of specialized fluoroelastomer (FKM) compounds for applications in rubber seals and gaskets. The influence of a commercially available perfluoropolyether (PFPE) plasticizer on the printability of a control FKM rubber compound was studied using a custom-designed ram material extruder, Additive Ram Material Extruder (ARME), for printing fully compounded thermoset elastomers. The plasticizer’s effectiveness was assessed based on its ability to address challenges such as high compound viscosity and post-print shrinkage, as well as its impact on interlayer adhesion. The addition of the PFPE plasticizer significantly reduced the FKM compound’s viscosity (by 70%) and post-print shrinkage (by 65%). While the addition of the plasticizer decreased the tensile strength of the control compound, specimens printed with the plasticized FKM retained 34% of the tensile strength of compression-molded samples, compared to only 23% for the unplasticized compound. Finally, the feasibility of seals and gaskets manufacturing using both conventional and unconventional additive manufacturing (AM) approaches was explored. A hybrid method combining AM and soft tooling for compression molding emerged as the optimal method for seal and gasket fabrication. Full article

Elastin-like polypeptides (ELPs) are popular biomaterials due to their reversible, temperature-dependent phase separation and their tunability, which is achievable by evolving procedures in recombinant technology. In particular, reverse direction ligation by plasmid reconstruction (PRe-RDL) is the predominant cloning technique used to generate ELPs of varying lengths. Pre-RDL provides precise control over the number of (VPGXG)_n repeat units in an ELP due to the selection of type IIS restriction enzyme (REs) sites in the reconstructed pET expression plasmid, which is a low-to-medium copy number plasmid. While Pre-RDL can be used to seamlessly repeat essentially any gene sequence and overcome limitations of previous cloning practices, we modified the Pre-RDL technique, where a high copy number plasmid (pBluescript II SK(+)—using a new library of type IIS REs) was used instead of a pET plasmid. The modified technique successfully produced a diblock ELP gene of 240 pentapeptide repeats from 30 pentapeptide “monomers” composed of alanine, tyrosine, and leucine X residues. This study found that the large, GC-rich ELP gene compromised plasmid yields in pBluescript II SK(+) and favored higher plasmid yields in the pET19b expression plasmid. Additionally, the BL21 E. coli strain expression consistently provided a higher transformation efficiency and higher plasmid yield than the high cloning efficiency strain TOP10 E. coli. We hypothesize that the plasmid/high GC gene ratio may play a significant role in these observations, and not the total plasmid size or the total plasmid GC content. While expression of the final gene resulted in a diblock ELP with a phase separation temperature of 34.5 °C, future work will need to investigate RDL techniques in additional plasmids to understand the primary driving factors for improving yields of plasmids with large ELP-encoding genes. Full article