Predicting Natural Evolution in the RBD Region of the Spike Glycoprotein of SARS-CoV-2 by Machine Learning.

Machine learning (ML) is a key focus in predicting protein mutations and aiding directed evolution. Research on potential virus variants is crucial for vaccine development. In this study, the machine learning software PyPEF was employed to conduct mutation analysis within the receptor-binding domain (RBD) of the Spike glycoprotein of SARS-CoV-2. Over 48,960,000 variants were predicted. Eight prospective variants that could surface in the future underwent modeling and molecular dynamics simulations. The study forecasts that the latest variant, ISOY2P5O1, may potentially emerge around 17 November 2023, with an approximate window of uncertainty of ±22 days. The ISOY8P5O2 variant displayed an increased binding capacity in the dry assay, with a total predicted binding energy of -110.306 kcal/mol. This represents an 8.25% enhancement in total binding energy compared to the original SARS-CoV-2 strain discovered in Wuhan (-101.892 kcal/mol). Reverse research confirmed the structural significance of mutation sites using ML models, particularly in the context of protein folding. The study validated regression methods (SVR, RF, and PLS) with different data structures. This study investigates the effectiveness of the "ML-Guided Design Correctly Predicts Combinatorial Effects Strategy" compared to the "ML-Guided Design Correctly Predicts Natural Evolution Prediction Strategy". To enhance machine learning, we created a timestamping algorithm and two auxiliary programs using advanced techniques to rapidly process extensive data, surpassing batch sequencing capabilities. This study not only advances machine learning in guiding protein evolution but also holds potential for forecasting future viruses and vaccine development.

Multistage Porous Carbon Derived from Enzyme-Treated Waste Walnut Green Husk and Polyethylene Glycol for Phase Change Energy Storage.

The thermal storage performance, cost, and stability of phase-change materials (PCMs) are critical factors influencing their application in the field of thermal energy storage. Porous carbon, with its excellent support, thermal conductivity, and energy storage properties, is considered one of the most promising support matrix materials. However, the simple and efficient synthesis of high-performance and highly active bio-based materials under mild conditions still faces challenges. In our work, a novel method for preparing new functional composite phase-change materials based on enzyme treatment technology and using waste walnut green husk biomass and polyethylene glycol as raw materials was developed. The enzymatic treatment method exposes the internal structure of the walnut green husk, followed by the adjustment of the calcination temperature to increase the adsorption sites of the biochar, thereby stabilizing polyethylene glycol (PEG). The porous properties of walnut green husk biochar effectively regulate the phase-change behavior of polyethylene glycol. In the biochar carbonized at 600 °C, the PEG loading reached 72.09%, and the absorption heat of the solid-solid phase-change material (SSPCM) reached 194.76 J g-1. This work not only enriches the application of biomass in heat storage but also demonstrates the broad prospects of SSPCMs in solar thermal utilization.

Engineered ß-glycosidase from Hyperthermophilic Sulfolobus solfataricus with Improved Rd-hydrolyzing Activity for Ginsenoside Compound K Production.

Hyperthermophilic Sulfolobus solfataricus ß-glycosidase (SS-ßGly), with higher stability and activity than mesophilic enzymes, has potential for industrial ginsenosides biotransformation. However, its relatively low ginsenoside Rd-hydrolyzing activity limits the production of pharmaceutically active minor ginsenoside compound K (CK). In this study, first, we used molecular docking to predict the key enzyme residues that may hypothetically interact with ginsenoside Rd. Then, based on sequence alignment and alanine scanning mutagenesis approach, key variant sites were identified that might improve the enzyme catalytic efficiency. The enzyme catalytic efficiency (kcat/Km) and substrate affinity (Km) of the N264D variant enzyme for ginsenoside Rd increased by 60% and decreased by 17.9% compared with WT enzyme, respectively, which may be due to a decrease in the binding free energy (∆G) between the variant enzyme and substrate Rd. In addition, Markov state models (MSM) analysis during the whole 1000-ns MD simulations indicated that altering N264 to D made the variant enzyme achieve a more stable SS-ßGly conformational state than the wild-type (WT) enzyme and corresponding Rd complex. Under identical conditions, the relative activities and the CK conversion rates of the N264D enzyme were 1.7 and 1.9 folds higher than those of the WT enzyme. This study identified an excellent hyperthermophilic ß-glycosidase candidate for industrial biotransformation of ginsenosides.

Exploring effective ways to increase reliable positive samples for machine learning-based urban waterlogging susceptibility assessments.

Machine learning (ML)-based urban waterlogging susceptibility studies suffer from class imbalance, as fewer positive samples are generally available than potential negative samples. Few studies have considered optimizing the results by improving the quality of training samples. To address this issue, we explored effective approaches to reliably increase the numbers of positive samples for such studies. The Synthetic Minority Over-Sampling Technique (SMOTE) and Optimized Seed Spread Algorithm (OSSA), representative of oversampling (synthesizing new samples based on the feature space) and physical (simulating potential inundated area based on the mechanisms of water flow) approaches, respectively, were employed to increase the number of positive samples. Waterlogging in Shenzhen was selected as a case study using eight selected spatial variables. An elaborate experiment was conducted to compare the quality of added samples based on the classifiers' performance and accuracy of waterlogging susceptibility maps (WSMs). The results indicated that (1) the performance of classifiers generated with SMOTE was worse than the original samples, while the use of OSSA improved the trained classifiers, and (2) the accuracy of WSMs was not improved with SMOTE but increased markedly with OSSA. These results may be driven by the diversity of information and features of the added samples. This study indicates the use of SMOTE fails to synthesize reliable samples when applied to waterlogging analysis in Shenzhen, whereas an effective solution for generating reliable positive samples is to use OSSA that simulates the potential submerged regions based on the mechanisms of disaster occurrence and spread.

Synthetic strategies to access silacycles.

In comparison with all-carbon parent compounds, the incorporation of Si-element into carboskeletons generally endows the corresponding sila-analogues with unique biological activity and physical-chemical properties. Silacycles have recently shown promising application potential in biological chemistry, pharmaceuticals industry, and material chemistry. Therefore, the development of efficient methodology to assemble versatile silacycles has aroused increasing concerns in the past decades. In this review, recent advances in the synthesis of silacycle-system are briefly summarized, including transition metal-catalytic and photocatalytic strategies by employing arylsilanes, alkylsilane, vinylsilane, hydrosilanes, and alkynylsilanes, etc. as starting materials. Moreover, a clear presentation and understanding of the mechanistic aspects and features of these developed reaction methodologies have been high-lighted.

Remote C-H Olefination of Heterocyclic Biaryls Enabled by Reversibly Bound Templates.

Remote C-H functionalization of heterocyclic biaryls will be of great importance in synthesis and medicinal chemistry. Through adjusting the geometric relationship of the directing atom and target C-H bonds, two new catalytic templates have been developed to enable the functionalization of the more hindered ortho-C-H bonds of heterobiaryls bearing directing heteroatom at the meta- or para-positions, affording unprecedented site-selectivity. The use of template chaperone also overcomes product inhibition and renders the directing templates catalytic. The utility of this protocol was demonstrated by olefination of heterocyclic biaryls with various substituents, overriding conventional steric and electronic effects. These ortho-C-H olefinated heterobiaryls are sterically hindered and can often be challenging to prepare through aryl-aryl coupling reactions.

ZNF219, a novel transcriptional repressor, inhibits transcription of the prototype foamy virus by interacting with the viral LTR promoter.

Prototype foamy virus (PFV) is an ancient retrovirus that infects humans with persistent latent infections and non-pathogenic consequences. Lifelong latent PFV infections can be caused by restrictive factors in the host. However, the molecular mechanisms underlying host cell regulation during PFV infection are not fully understood. The aim of the study was to investigate whether a zinc finger protein (ZFP), ZNF219, as a transcription factor, can regulate the transcriptional activity of the viral promoter. Here, using transcriptome sequencing, we found that ZNF219, is downregulated in PFV infected cells and that ZNF219 suppresses viral replication by targeting the viral 5'LTR promoter region to repress its transcription. We also found that PFV infection induced abnormal expression of miRNAs targeting the ZNF219-3'UTR to downregulate ZNF219 expression. These findings indicated that ZNF219 may be a potent antiviral factor for suppressing PFV infection, and may shed light on the mechanism of virus-host interactions.

Analyses of the Cellular Interactions between the Ossification of Collagen-Based Barrier Membranes and the Underlying Bone Defects.

Barrier membranes are an essential tool in guided bone Regeneration (GBR), which have been widely presumed to have a bioactive effect that is beyond their occluding and space maintenance functionalities. A standardized calvaria implantation model was applied for 2, 8, and 16 weeks on Wistar rats to test the interactions between the barrier membrane and the underlying bone defects which were filled with bovine bone substitute materials (BSM). In an effort to understand the barrier membrane's bioactivity, deeper histochemical analyses, as well as the immunohistochemical detection of macrophage subtypes (M1/M2) and vascular endothelial cells, were conducted and combined with histomorphometric and statistical approaches. The native collagen-based membrane was found to have ossified due to its potentially osteoconductive and osteogenic properties, forming a "bony shield" overlying the bone defects. Histomorphometrical evaluation revealed the resorption of the membranes and their substitution with bone matrix. The numbers of both M1- and M2-macrophages were significantly higher within the membrane compartments compared to the underlying bone defects. Thereby, M2-macrophages significantly dominated the tissue reaction within the membrane compartments. Statistically, a correlation between M2-macropahges and bone regeneration was only found at 2 weeks post implantationem, while the pro-inflammatory limb of the immune response correlated with the two processes at 8 weeks. Altogether, this study elaborates on the increasingly described correlations between barrier membranes and the underlying bone regeneration, which sheds a light on the understanding of the immunomodulatory features of biomaterials.

Suitability of R. pulmo Jellyfish-Collagen-Coated Well Plates for Cytocompatibility Analyses of Biomaterials.

Cytocompatibility analyses of new implant materials or biomaterials are not only prescribed by the Medical Device Regulation (MDR), as defined in the DIN ISO Norm 10993-5 and -12, but are also increasingly replacing animal testing. In this context, jellyfish collagen has already been established as an alternative to mammalian collagen in different cell culture conditions, but a lack of knowledge exists about its applicability for cytocompatibility analyses of biomaterials. Thus, the present study was conducted to compare well plates coated with collagen type 0 derived from Rhizostoma pulmo with plates coated with bovine and porcine collagen. The coated well plates were analysed in vitro for their cytocompatibility, according to EN ISO 10993-5/-12, using both L929 fibroblasts and MC3T3 pre-osteoblasts. Thereby, the coated well plates were compared, using established materials as positive controls and a cytotoxic material, RM-A, as a negative control. L929 cells exhibited a significantly higher viability (#### p < 0.0001), proliferation (## p < 0.01), and a lower cytotoxicity (## p < 0.01 and # p < 0.05)) in the Jellagen® group compared to the bovine and porcine collagen groups. MC3T3 cells showed similar viability and acceptable proliferation and cytotoxicity in all collagen groups. The results of the present study revealed that the coating of well plates with collagen Type 0 derived from R. pulmo leads to comparable results to the case of well plates coated with mammalian collagens. Therefore, it is fully suitable for the in vitro analyses of the cytocompatibility of biomaterials or medical devices.

High-Throughput Screening and Directed Evolution of Methionine Adenosyltransferase from Escherichia coli.

S-adenosyl-L-methionine (SAM) is the active form of methionine, which participates in various metabolic reactions and plays a vital role. It is mainly used as a precursor by three key metabolic pathways: trans-methylation, trans-sulfuration, and trans-aminopropylation. Methionine adenosyltransferase (MAT) is the only enzyme to produce SAM from methionine and ATP. However, there is no efficient and accurate method for high-throughput detection of SAM, which is the major obstacles of directed evolution campaigns for MAT. Herein, we established a colorimetric method for directed evolution of MAT based on detecting SAM by using glycine oxidase and glycine/sarcosine N-methyltransferase enzyme. Screening of MAT libraries revealed variant I303V/Q22R with 2.13-fold improved activity towards SAM in comparison to the wild type. Molecular dynamic simulation indicates that the loops more flexible and more conducive to SAM release.

Barrier Membranes for Guided Bone Regeneration (GBR): A Focus on Recent Advances in Collagen Membranes.

Guided bone regeneration (GBR) has become a clinically standard modality for the treatment of localized jawbone defects. Barrier membranes play an important role in this process by preventing soft tissue invasion outgoing from the mucosa and creating an underlying space to support bone growth. Different membrane types provide different biological mechanisms due to their different origins, preparation methods and structures. Among them, collagen membranes have attracted great interest due to their excellent biological properties and desired bone regeneration results to non-absorbable membranes even without a second surgery for removal. This work provides a comparative summary of common barrier membranes used in GBR, focusing on recent advances in collagen membranes and their biological mechanisms. In conclusion, the review article highlights the biological and regenerative properties of currently available barrier membranes with a particular focus on bioresorbable collagen-based materials. In addition, the advantages and disadvantages of these biomaterials are highlighted, and possible improvements for future material developments are summarized.

In Vivo Analysis of the Regeneration Capacity and Immune Response to Xenogeneic and Synthetic Bone Substitute Materials.

Although various studies have investigated differences in the tissue reaction pattern to synthetic and xenogeneic bone substitute materials (BSMs), a lack of knowledge exists regarding the classification of both materials based on the DIN ISO 10993-6 scoring system, as well as the histomorphometrical measurement of macrophage subtypes within their implantation beds. Thus, the present study was conducted to analyze in vivo responses to both xenogeneic and synthetic bone substitute granules. A standardized calvaria implantation model in Wistar rats, in combination with established scoring, histological, histopathological, and histomorphometrical methods, was conducted to analyze the influence of both biomaterials on bone regeneration and the immune response. The results showed that the application of the synthetic BSM maxresorb® induced a higher pro-inflammatory tissue response, while the xenogeneic BSM cerabone® induced a higher anti-inflammatory reaction. Additionally, comparable bone regeneration amounts were found in both study groups. Histopathological scoring revealed that the synthetic BSM exhibited non-irritant scores at all timepoints using the xenogeneic BSM as control. Overall, the results demonstrated the biocompatibility of synthetic BSM maxresorb® and support the conclusion that this material class is a suitable alternative to natural BSM, such as the analyzed xenogeneic material cerabone®, for a broad range of indications.

Effect of Text Messaging Parents of School-Aged Children on Outdoor Time to Control Myopia: A Randomized Clinical Trial.

Importance: Myopia in school-aged children is a public health issue worldwide; consequently, effective interventions to prevent onset and progression are required. Objective: To investigate whether SMS text messages to parents increase light exposure and time outdoors in school-aged children and provide effective myopia control. Design, Setting, and Participants: This randomized clinical trial was conducted in China from May 2017 to May 2018, with participants observed for 3 years. Of 528 965 primary school-aged children from Anyang, 3113 were randomly selected. Of these, 268 grade 2 schoolchildren were selected and randomly assigned to SMS and control groups. Data were analyzed from June to December 2021. Interventions: Parents of children in the SMS group were sent text messages twice daily for 1 year to take their children outdoors. All children wore portable light meters to record light exposure on 3 randomly selected days (2 weekdays and 1 weekend day) before and after the intervention. Main Outcomes and Measures: The co-primary outcomes were change in axial length (axial elongation) and change in spherical equivalent refraction (myopic shift) from baseline as measured at the end of the intervention and 3 years later. A secondary outcome was myopia prevalence. Results: Of 268 grade 2 schoolchildren, 121 (45.1%) were girls, and the mean (SD) age was 8.4 (0.3) years. Compared with the control group, the SMS intervention group demonstrated greater light exposure and higher time outdoors during weekends, and the intervention had significant effect on axial elongation (coefficient, 0.09; 95% CI, 0.02-0.17; P = .01). Axial elongation was lower in the SMS group than in the control group during the intervention (0.27 mm [95% CI, 0.24-0.30] vs 0.31 mm [95% CI, 0.29-0.34]; P = .03) and at year 2 (0.39 mm [95% CI, 0.35-0.42] vs 0.46 mm [95% CI, 0.42-0.50]; P = .009) and year 3 (0.30 mm [95% CI, 0.27-0.33] vs 0.35 mm [95% CI, 0.33-0.37]; P = .005) after the intervention. Myopic shift was lower in the SMS group than in the control group at year 2 (-0.69 diopters [D] [95% CI, -0.78 to -0.60] vs -0.82 D [95% CI, -0.91 to -0.73]; P = .04) and year 3 (-0.47 D [95% CI, -0.54 to -0.39] vs -0.60 D [95% CI, -0.67 to -0.53]; P = .01) after the intervention, as was myopia prevalence (year 2: 38.3% [51 of 133] vs 51.1% [68 of 133]; year 3: 46.6% [62 of 133] vs 65.4% [87 of 133]). Conclusions and Relevance: In this randomized clinical trial, SMS text messages to parents resulted in lower axial elongation and myopia progression in schoolchildren over 3 years, possibly through increased outdoor time and light exposure, showing promise for reducing myopia prevalence. Trial Registration: Chinese Clinical Trial Registry Identifier: ChiCTR-IOC-17010525.

Feasible Cluster Model Method for Simulating the Redox Potentials of Laccase CueO and Its Variant.

Laccases are regarded as versatile green biocatalysts, and recent scientific research has focused on improving their redox potential for broader industrial and environmental applications. The density functional theory (DFT) quantum mechanics approach, sufficiently rigorous and efficient for the calculation of electronic structures, is conducted to better comprehend the connection between the redox potential and the atomic structural feature of laccases. According to the crystal structure of wild type laccase CueO and its variant, a truncated miniature cluster model method was established in this research. On the basic of thermodynamic cycle, the overall Gibbs free energy variations before and after the one-electron reduction were calculated. It turned out that the trends of redox potentials to increase after variant predicted by the theoretical calculations correlated well with those obtained by experiments, thereby validating the feasibility of this cluster model method for simulating the redox potentials of laccases.

Directed Self-Assembly of Heterologously Expressed Hagfish EsTKα and EsTKγ for Functional Hydrogel.

Hagfish slime proteins have long been considered useful due to their potential applications in novel green, environmental, and functional bionic materials. The two main component proteins in the slime thread of hagfish, (opt)EsTKα and (opt)EsTKγ, were used as raw materials. However, the methods available to assemble these two proteins are time- and labor-intensive. The conditions affecting protein self-assembly, such as the pH of the assembly buffer, protein concentration, and the protein addition ratio, were the subject of the present research. Through a series of tests, the self-assembly results of a variety of assembly conditions were explored. Finally, a simplified protein self-assembly method was identified that allows for simple, direct assembly of the two proteins directly. This method does not require protein purification. Under the optimal assembly conditions obtained by exploration, a new gel material was synthesized from the hagfish protein through self-assembly of the (opt)EsTKα and (opt)EsTKγ. This assembly method has the benefits of being a simple, time-saving, and efficient. The self-assembled protein gel products were verified by SDS polyacrylamide gel electrophoresis (SDS-PAGE) and contained (opt)EsTKα and (opt)EsTKγ proteins. Scanning electron microscopy (SEM) was used to investigate the self-assembled protein gel after freeze-drying, and it was observed that the self-assembled protein formed a dense, three-dimensional porous network structure, meaning that it had good water retention. Evaluation of the gel with atomic force microscopy (AFM) indicated that the surface of the protein fiber skeleton show the network-like structure and relatively smooth. Characterization by circular dichroism (CD) and Fourier transform infrared spectroscopy (FT-IR) demonstrated that the two proteins were successfully assembled, and that the assembled protein had a secondary structure dominated by α-helices. The rheological properties of the self-assembled products were tested to confirm that they were indeed hydrogel property.

Functionalization of Se-Te Nanorods with Au Nanoparticles for Enhanced Anti-Bacterial and Anti-Cancer Activities.

The use of medical devices for therapeutic and diagnostic purpose is globally increasing; however, bacterial colonization on therapeutic devices can occur, causing severe infections in the human body. It has become an issue for public health. It is necessary to develop a nanomaterial based on photothermal treatment to kill toxic bacterial strains. Appropriately, high photothermal conversion and low-cost powerful photothermal agents have been investigated. Recently, gold nanocomposites have attracted great interest in biological applications. Here, we prepared rod-shaped Se-Te@Au nanocomposites of about 200 nm with uniform shape and surface-coated with gold nanoparticles for the first time showing high anti-bacterial and anti-cancer activities. Se-Te@Au showed proper structural consistency and natural resistance to bacterial and cancer cells. The strong absorption and high photothermal conversion efficacy made it a good photothermal agent material for the photothermal treatment of bacterial and cancer cells. The Se-Te@Au rod showed excellent anti-bacterial efficacy against Gram-negative Escherichia coli and Gram-positive Staphylococcus aureus, with highest recorded inhibition zones of 25 ± 2 mm and 22 ± 2 mm, respectively. More than 99% of both types of strains were killed after 5 min with a near-infrared (NIR) laser at the very low concentration of 48 µg/mL. The Se-Te@Au rod's explosion in HeLa cells was extensively repressed and demonstrated high toxicity at 100 µg/mL for 5 min when subjected to an NIR laser. As a result of its high photothermal characteristics, the exceptional anti-bacterial and anti-cancer effects of the Se-Te@Au rod are considerably better than those of other methods previously published in articles. This study could open a new framework for sterilization applications on the industrial level.

Process analysis of microplastic degradation using activated PMS and Fenton reagents.

The environmental degradation and physical aging of microplastics (MP) caused by oxidative stress have not been thoroughly elucidated. In this study, we used different oxidative agents (Fe2+-activated peroxymonosulfate and Fenton reagents) that can form free radicals to study the degradation mechanisms of nylon 6 (PA6) and polystyrene (PS) MPs. After 4 cycles of treatment, mass losses of 25.6% and 22.1% were obtained with PA6 and PS MPs, respectively. Scanning electron microscopy (SEM), Fourier-transform infrared (FTIR) spectroscopy, Raman spectroscopy, and X-ray photoelectron spectroscopy (XPS) were employed to confirm the MP oxidation, and polymer chain scissions. FTIR data indicated the existence of oxygen-containing chemical groups resulting from MPs oxidation, and chain scissions, such as CO, C-O, and O-CO. Raman spectroscopy suggested the presence of exposed aromatic groups, and weakening of the relative intensity of C-H related to the oxidation, and chain scission of the MPs. SEM imaging revealed visible morphological changes on the surface of MPs as a result of degradation. XPS indicated that the O/C ratio could be used as an indicator for the degree of MPs oxidation. By analyzing the degradation products with surface-enhanced Raman scattering (SERS) and gas chromatography-mass spectrometry, low-molecular-weight alkanes, alcohols, aldehydes, carboxylic acids derived from the derivatization of alcohols, were detected. These findings confirmed the advantage of using multiple analytic methods in tandem to evaluate the degradation of environmental MPs.

The performance of an integrated model including retinal information in predicting childhood hypertension.

BACKGROUND: The objective of this study was to examine the association of an integrated model (composed of retinal arteriolar caliber, height, and sex) with blood pressure (BP) among a group of Chinese children, and assess the predictive value of the integrated model for childhood hypertension. METHODS: This study included 1460 candidates aged 12.634 ± 0.420 years. Height, weight, waist circumference, and BP were obtained and ophthalmological measurements were taken. The computer-imaging program (IVAN, University of Wisconsin, Madison, WI) was used to measure calibers of retinal vessels. Receiver-operating characteristic curve (ROC) analyses were performed to assess the accuracy of the integrated model as a diagnostic test of elevated BP in children. RESULTS: The accuracy of the integrated model (assessed by area under the curve) for identifying elevated BP was 0.777 (95% confidence interval: 0.742-0.812). The optimal threshold of the integrated model for defining hypertension was 0.153, and the calculation formula for the specific predictive risk was: Logit (p/1 - p) = -5.666 - 0.261 × retinal arteriolar caliber + 0.945 × sex + 0.438 × height. In identifying elevated BP, the sensitivity and specificity were 0.711 and 0.736, respectively. CONCLUSIONS: The model containing eye message is a comprehensive and relatively effective index to identify elevated BP in 12-year-old children, which can offer assistance to further understand childhood microcirculation disease. IMPACT: We firstly incorporated retinal vascular diameter, sex, and height into one integrated model to identify hypertension in 12-year-old children. The current discrimination of hypertension in children is difficult. There have been some studies to simplify the diagnosis of children's hypertension, but they were limited to anthropometric measurements. We proposed a composed model containing microcirculation information to predict childhood hypertension. Based on the knowledge that microcirculation is not only a means to study the manifestations but also early pathogenic correlates of hypertension, the combined model containing microcirculation message as a method may provide new insights into the diagnosis of childhood hypertension.

Modulating the Coupling Efficiency of P450 BM3 by Controlling Water Diffusion through Access Tunnel Engineering.

Cytochromes P450 have gained much interest for their broad substrate scope in the catalysis of oxidation reactions for pharmaceuticals, plastics, and hormones. However, achieving high coupling efficiency by the engineering of P450s is still a big challenge. The presence of extra water around the active site is deemed to be related to uncoupling. In this study, the access tunnels of P450 BM3 from Bacillus megaterium are engineered to control water access from bulk solvent to the active site. Nine residues located in tunnels are investigated by site-saturation mutagenesis to reduce water diffusion, thereby improving the coupling efficiency. The recombined variant N319L/T411V/T436A shows improved coupling efficiency (from 31.2 % to 52.6 %). Tunnel polarity analysis and molecular dynamics simulation further indicate that reduced water molecules around the active site lead to higher coupling efficiency. Overall, this study provides valuable insight on improving coupling efficiency by controlling water diffusion through tunnel engineering.

Application of 6S and PDCA cycle lean management in nursing management of orthopedic trauma surgery / 中华创伤杂志

Objective:To explore the application effect of 6S and PDCA cycle lean management in nursing management of orthopaedic trauma surgery.Methods:A retrospective cohort study was conducted to analyze the clinical data of 80 patients with orthopedic trauma admitted to University-Town Hospital of Chongqing Medical University from January 2020 to November 2021, including 51 males and 29 females; aged 23-68 years [(44.5±9.2)years]. According to the order of operation date, 40 patients from January 2021 to November 2021 received routine management plus 6S and PDCA cycle lean management (lean management group), and 40 patients from January 2020 to December 2020 received routine surgical nursing management (routine management group). The nursing quality score in operating room (instrument and equipment management, surgical instrument preparation, surgical coordination, disinfection and isolation), qualified rate of infection monitoring in operating room (air in operating room, hand hygiene of medical staffs and sterile articles in operating room), surgeons′ satisfaction with operating nurses (circulating nurses and scrub nurses), self- rating anxiety scale (SAS) of patients before operation, incidence of complications at postoperative 12 days and patients′ satisfaction with nurses in operating room at postoperative 2-3 days.Results:The four dimensions of the nursing quality score in operating room in lean management group were (27.8±1.5)points, (18.1±1.1)points, (26.6±1.5)points and (18.1±0.8)points, significantly higher than (23.7±2.3)points, (14.6±1.1)points, (22.4±1.8)points and (13.7±1.1)points in routine management group (all P<0.01). The three dimensions of qualified rate of infection monitoring in operating room in lean management group were all 100%, showing no significant differences in routine management group (75%, 100%, 100%) (all P>0.05). Surgeons′ satisfaction with circulating nurses and scrub nurses in lean management group was 90% and 85%, higher than 73% and 65% in routine management group (all P<0.05). Preoperative SAS in lean management group was (32.5±8.2)points, significantly lower than (54.6±10.7)points in routine management group ( P<0.01). The incidence of postoperative complications in lean management group was 0, significantly lower than 15%(6/40) in routine management group ( P<0.05). Patients′ satisfaction with nurses in operating room in lean management group was 95% (38/40), significantly higher than 78%(31/40) in routine management group ( P<0.05). Conclusion:6S and PDCA cycle lean management can effectively improve the nursing quality of orthopedic trauma surgery, reduce patients′ anxiety and postoperative complications, and improve surgeons′ and patients′ satisfaction, which is worthy of further clinical application.