Multivariate Silicification-Assisted Single Enzyme Structure Augmentation for Improved Enzymatic Activity-Stability Trade-Off.

The ability to finely tune/balance the structure and rigidity of enzymes to realize both high enzymatic activity and long-term stability is highly desired but highly challenging. In this work, we propose a new concept of silica-enzyme, referred to as "silicazyme", where solid inorganic silica was controlled hybridization with fragile enzyme under moderate condition at single-enzyme level, realizing simultaneous structure augmentation, long-term stability, and high enzymatic activity preservation. A multivariate silicification approach was utilized and occurred around individual enzymes to allow conformal coating. To realize a high activity-stability trade-off the structure flexibility/rigidity of silicazyme was optimized by a component-adjustment-ternary (CAT) plot method. Moreover, the multivariate organosilica frameworks bring great advantages including surface microenvironment adjustability, reversible modification capability, and functional extensibility through the rich chemistry of silica. Overall silicazymes represent a new class of enzymes that promise to broaden their utilization in catalysis, separations, and nanomedicine.

Carboxymethyl cellulose-based multifunctional film integrated with polyphenol-rich extract and carbon dots from coffee husk waste for active food packaging applications.

Sustainable carboxymethyl cellulose (CMC)-based active composite films were developed through the addition of polyphenol-rich extract from coffee husk (CHE) and carbon dots (CDs) prepared using the biowaste residue of CHE extraction. The influences of various CDs contents on the physicochemical and functional characteristics of composite films have been researched. The 6% (w/w) CHE and 3% (w/w) CDs were uniformly dispersed within the CMC matrix to produce a homogenous film with enhanced mechanical properties. The CMC/CHE/CDs3% film exhibited outstanding UV-light blocking, improved water and gas barriers, potent antioxidant activity with above 95% DPPH and ABTS scavenging rates, and effective antibacterial capabilities against L. monocytogenes and E. coli. The food packaging experiment demonstrated that this active composite film slowed the rotting of fresh-cut apples and extended their shelf-life to 7 days at 4 °C storage. Therefore, the obtained multifunctional film showed promise as an environmentally friendly food packaging material.

Development and Validation of a Pathomics Model Using Machine Learning to Predict CXCL8 Expression and Prognosis in Head and Neck Cancer.

OBJECTIVES: The necessity to develop a method for prognostication and to identify novel biomarkers for personalized medicine in patients with head and neck squamous cell carcinoma (HNSCC) cannot be overstated. Recently, pathomics, which relies on quantitative analysis of medical imaging, has come to the forefront. CXCL8, an essential inflammatory cytokine, has been shown to correlate with overall survival (OS). This study examined the relationship between CXCL8 mRNA expression and pathomics features and aimed to explore the biological underpinnings of CXCL8. METHODS: Clinical information and transcripts per million mRNA sequencing data were obtained from The Cancer Genome Atlas (TCGA)-HNSCC dataset. We identified correlations between CXCL8 mRNA expression and patient survival rates using a Kaplan-Meier survival curve. A retrospective analysis of 313 samples diagnosed with HNSCC in the TCGA database was conducted. Pathomics features were extracted from hematoxylin and eosin-stained images, and then the minimum redundancy maximum relevance, with recursive feature elimination (mRMR-RFE) method was applied, followed by screening with the logistic regression algorithm. RESULTS: Kaplan-Meier curves indicated that high expression of CXCL8 was significantly associated with decreased OS. The logistic regression pathomics model incorporated 16 radiomics features identified by the mRMR-RFE method in the training set and demonstrated strong performance in the testing set. Calibration plots showed that the probability of high gene expression predicted by the pathomics model was in good agreement with actual observations, suggesting the model's high clinical applicability. CONCLUSION: The pathomics model of CXCL8 mRNA expression serves as an effective tool for predicting prognosis in patients with HNSCC and can aid in clinical decision-making. Elevated levels of CXCL8 expression may lead to reduced DNA damage and are associated with a pro-inflammatory tumor microenvironment, offering a potential therapeutic target.

Synthetic Biohybrids of Red Blood Cells and Cascaded-Enzymes@ Metal-Organic Frameworks for Hyperuricemia Treatment.

Hyperuricemia, caused by an imbalance between the rates of production and excretion of uric acid (UA), may greatly increase the mortality rates in patients with cardiovascular and cerebrovascular diseases. Herein, for fast-acting and long-lasting hyperuricemia treatment, armored red blood cell (RBC) biohybrids, integrated RBCs with proximal, cascaded-enzymes of urate oxidase (UOX) and catalase (CAT) encapsulated within ZIF-8 framework-based nanoparticles, have been fabricated based on a super-assembly approach. Each component is crucial for hyperuricemia treatment: 1) RBCs significantly increase the circulation time of nanoparticles; 2) ZIF-8 nanoparticles-based superstructure greatly enhances RBCs resistance against external stressors while preserving native RBC properties (such as oxygen carrying capability); 3) the ZIF-8 scaffold protects the encapsulated enzymes from enzymatic degradation; 4) no physical barrier exists for urate diffusion, and thus allow fast degradation of UA in blood and neutralizes the toxic by-product H2 O2 . In vivo results demonstrate that the biohybrids can effectively normalize the UA level of an acute hyperuricemia mouse model within 2 h and possess a longer elimination half-life (49.7 ± 4.9 h). They anticipate that their simple and general method that combines functional nanomaterials with living cell carriers will be a starting point for the development of innovative drug delivery systems.

Crosslinked fish scale gelatin/alginate dialdehyde functional films incorporated with carbon dots derived from pomelo peel waste for active food packaging.

A multifunctional and environmentally friendly composite film was developed by incorporating pomelo peel-derived carbon dots (PCDs) into a fish scale gelatin (FSG)/alginate dialdehyde (ADA) biopolymer matrix. ADA was used to reinforce the physicomechanical properties of the FSG film via Schiff base crosslinking. PCDs with strong antioxidant and antimicrobial activities were synthesized via a hydrothermal method. The effect of various PCDs content on the surface morphological, physicochemical, and functional characteristics of the composite films was investigated. The results showed that the introduction of PCDs into the FSG/ADA matrix effectively reinforced the mechanical performance, enhanced the water vapor and water resistance, increased UV-light blocking, conferred fluorescence properties, and improved the thermal properties of the composite films. Under 3 wt% PCDs content, the FSG/ADA/PCDs-3 % composite film not only presented significant antioxidant capacity with a radical scavenging rate of 91.71 % for DPPH and approximately 100 % for ABTS, but also exhibited excellent antimicrobial ability against bacteria and fungi. Results of a preservation experiment showed that the prepared FSG/ADA/PCDs-3 % film preserved the physiological qualities of strawberries post-harvest and extended their shelf-life to 7 days at room temperature. Overall, the fabricated FSG/ADA/PCDs composite films are promising for use in eco-friendly active food packaging.

Comparison of the Effects of Laparoscopic Surgery and Traditional Open Surgery on Stone Clearance, Laboratory Indexes and Life Quality in Patients with Renal Calculi.

Objective: A case-control study was conducted to determine the effectiveness of laparoscopic surgery and traditional open surgery on stone clearance, laboratory indexes, and life quality in patients with renal calculi. Methods: During March 2017 to March 2022, 272 patients with complex renal calculi (CRC) cured in our hospital were assigned into control group (n = 136) and research group (n = 136) arbitrarily. The former accepted traditional open surgery, while the latter accepted laparoscopic surgery. The operation time, intraoperative blood loss, hospital stay, and time of getting out of bed were compared. The degree of postoperative incision pain was assessed by visual analogue scale (VAS). The life quality was assessed by the Comprehensive Assessment Questionnaire-74 (GQOL-74). The indexes of renal function and urine metabolism were measured. Then, the postoperative stone clearance rate and complications were calculated. Results: Operation time, blood loss intraoperatively, time out of bed, and hospitalization were all remarkably reduced in the research group, and the difference was statistically significant (P < 0.05). The complete stone clearance rates in study and control cohorts were 75.73% and 63.24%, respectively. The VAS scores were lessened after the operation. Compared with the two groups, the VAS scores of the research group were remarkably lower at 1 to 2 weeks and 1 and 3 months after the operation, and the difference was statistically significant (P < 0.05). One week after operation, the levels of ß 2-microglobulin (ß 2-MG), N-acetyl-ß-glucosaminidase (NAG), and renal injury molecule-1 (kidney injury molecule-1, Kim-1) in the research group were remarkably lower. The levels of urinary ß 2-MG, NAG, and KIM-1 in the research group were remarkably lower, and the difference was statistically significant (P < 0.05). One week after operation, the levels of urinary oxalic acid, uric acid, and urinary calcium lessened averagely. The levels of urinary oxalic acid, uric acid, and urinary calcium in the research group were lower, and the difference was statistically significant (P < 0.05). The quality-of-life scores were compared. One week after the operation, the scores of physical function, psychological function, social function, and material function were all augmented, and the difference was statistically significant (P < 0.05). The incidence of complications was 9.56% and 2.21%, respectively. The incidence of complications in the research group was lower, and the difference was statistically significant (P < 0.05). Conclusion: Laparoscopic surgery is successful when treating CRC, which is superior to invasive surgery in postoperative complications, stone clearance rate, improvement of postoperative renal function, and life quality. It is one of the ideal treatment methods for CRC. However, the role of open surgery when treating CRC cannot be ignored. This needs to be further confirmed by large samples of randomized controlled trials.

Efficacy of Flexible Ureteroscopic Lithotripsy and Percutaneous Nephrolithotomy in the Treatment of Complex Upper Urinary Tract Nephrolithiasis.

Objective: A case-control study was conducted to compare the efficacy and prognostic factors of flexible ureteroscopic lithotripsy (FURL) and percutaneous nephrolithotomy (PCNL) when treating complex upper urinary tract renal calculi based on a retrospective cohort study. Methods: The study period was from October 2019 to December 2021. A retrospective study was carried out on 100 patients with complicated upper urinary tract nephrolithiasis who underwent surgery in the Urology Department of our hospital. They were assigned into two groups: FURL and PCNL groups. The intraoperative blood loss, operation time, hematuria duration, hospital stay, primary stone removal rate, incidence of intraoperative and postoperative complications, VAS score, level of inflammatory factors, and micturition function were compared. According to the postoperative prognosis, the patients were reassigned into two groups: good prognosis group (n = 38) and poor prognosis group (n = 106). The factors related to poor prognosis after FURL and PCNL were screened, and multivariate loglistic regression analysis was adopted to determine the risk factors. Results: The primary stone clearance rate in the PCNL group was significantly higher than that in the FURL group, and there was no significant difference in the incidence of intraoperative and postoperative complications between the two groups (P > 0.05). The intraoperative blood loss and the duration of hematuria in the PCNL group were significantly shorter than those in the FURL group, and the operation time and postoperative hospital stay in the FURL group were longer than those in the FURL group. The postoperative VAS score in the study group was significantly lower than that in the control group (P < 0.05). The levels of CRP, IL-1, TNF-α, and NF-κB in both groups decreased after operation, and the level of inflammatory factors in the PCNL group was significantly lower than that in the FURL group (P < 0.05). The indexes of IPSS and Q max in the PCNL group were significantly lower than those in the control group 3 months after operation. The index of micturition function in the PCNL group was significantly lower than that in the FURL group. Preoperative use of immunosuppressant, preoperative stone fever, positive preoperative urine culture, preoperative urinary leukocyte count ≥ 544 × L, intraoperative urinary opacity, and pus fur were significantly correlated with poor prognosis of ureteral patients (P < 0.05). Preoperative stone fever, high preoperative urinary leukocyte count, intraoperative urinary turbidity, and suppurative fur were independent risk factors for postoperative SIRS in patients with ureteral calculi. Conclusion: PCNL is effective when treating complex upper urinary tract renal calculi. Compared with FURL, PCNL can remarkably reduce intraoperative blood loss and hematuria duration, can enhance micturition function, and will not remarkably increase the incidence of intraoperative and postoperative complications, high safety. High white blood cell count in urine before operation, fever due to stone before operation, turbid urine, and purulent fur during operation are independent risk factors for postoperative adverse outcome in patients with complex upper urinary tract renal calculi. Patients should be fully treated before surgery.

Residues R1075, D1090, R1095, and C1130 Are Critical in ADAMTS13 TSP8-Spacer Interaction Predicted by Molecular Dynamics Simulation.

ADAMTS13 (A Disintegrin and Metalloprotease with Thrombospondin type 1 repeats, member 13) cleaves von Willebrand Factor (VWF) multimers to limit the prothrombotic function of VWF. The deficiency of ADAMTS13 causes a lethal thrombotic microvascular disease, thrombotic thrombocytopenic purpura (TTP). ADAMTS13 circulates in a "closed" conformation with the distal domain associating the Spacer domain to avoid off-target proteolysis or recognition by auto-antibodies. However, the interactions of the distal TSP8 domain and the Spacer domain remain elusive. Here, we constructed the TSP8-Spacer complex by a combination of homology modelling and flexible docking. Molecular dynamics simulation was applied to map the binding sites on the TSP8 or Spacer domain. The results predicted that R1075, D1090, R1095, and C1130 on the TSP8 domain were key residues that interacted with the Spacer domain. R1075 and R1095 bound exosite-4 tightly, D1090 formed multiple hydrogen bonds and salt bridges with exosite-3, and C1130 interacted with both exosite-3 and exosite-4. Specific mutations of exosite-3 (R568K/F592Y/R660K/Y661F/Y665F) or the four key residues (R1075A/D1090A/R1095A/C1130A) impaired the binding of the TSP8 domain to the Spacer domain. These results shed new light on the understanding of the auto-inhibition of ADAMTS13.

Characterization of the interactions of ADAMTS13 CUB1 domain to WT- and GOF-Spacer domain by molecular dynamics simulation.

Metalloprotease ADAMTS13 specifically cleaves VWF (von Willebrand Factor) to prevent excessive platelet aggregation and thrombus formation at the sites of vascular injury. To avoid non-specific cleavage, ADAMTS13 has the auto-inhibition effect in which the Spacer domain in N-terminal interacts with the CUB1 domain in C-terminal, resulting in decreased proteolytic activity. Previous studies reported that exosite-3 in the Spacer domain was a key binding site in the Spacer-CUB1 interaction. When exosite-3 was mutated (R660K/F592Y/R568K/Y661F/Y665F, GOF), the auto-inhibition of ADAMTS13 was disrupted and the enzymatic activity was markedly increased. However, the characteristics of the Spacer-CUB1 interaction is not fully understood. Here, we constructed the model of Spacer-CUB1 complex by homologous modeling and molecular docking to characterize the Spacer-CUB1 binding and predict key amino acid residues via molecular dynamics simulation. Our data showed that G607-S610 was a non-reported potential binding site in the Spacer domain; GOF mutation attenuated the formation of hydrogen bond between exosite-3 and the CUB1 domain; Residues E1231, R1251, L1258, D1259 and T1261 in the CUB1 domain might play an important role in the Spacer-CUB1 interaction. Our study advances the understanding of the structural basis of the auto-inhibition of ADAMTS13 and provides information about the key residues in the binding interface.

Chiral Phosphoric Acid-Catalyzed Remote Control of Axial Chirality at Boron-Carbon Bond.

The previously elusive catalytic enantioselective construction of axially chiral B-aryl-1,2-azaborines with a C-B stereogenic axis has been realized through a chiral phosphoric acid-catalyzed desymmetrization strategy reported herein. The electrophilic aromatic substitution reaction of 3,5-disubsituted phenols with diazodicarboxamides could afford these axially chiral structures in good efficiency with excellent enantiocontrol. The efficient long-range stereochemical control is achieved by multiple well-defined H-bonding interactions between chiral phosphoric acid and both substrates. Meanwhile, the reaction duration could be markedly shortened with weakly acidic N-H in 1,2-azaborine acting as H-bond donor. The scalability of the reaction and facile cleavage of the N-N bond in the product further demonstrated the practicality of this method.

Insights Into Immunothrombosis: The Interplay Among Neutrophil Extracellular Trap, von Willebrand Factor, and ADAMTS13.

Both neutrophil extracellular traps (NETs) and von Willebrand factor (VWF) are essential for thrombosis and inflammation. During these processes, a complex series of events, including endothelial activation, NET formation, VWF secretion, and blood cell adhesion, aggregation and activation, occurs in an ordered manner in the vasculature. The adhesive activity of VWF multimers is regulated by a specific metalloprotease ADAMTS13 (a disintegrin and metalloproteinase with thrombospondin type 1 motifs, member 13). Increasing evidence indicates that the interaction between NETs and VWF contributes to arterial and venous thrombosis as well as inflammation. Furthermore, contents released from activated neutrophils or NETs induce the reduction of ADAMTS13 activity, which may occur in both thrombotic microangiopathies (TMAs) and acute ischemic stroke (AIS). Recently, NET is considered as a driver of endothelial damage and immunothrombosis in COVID-19. In addition, the levels of VWF and ADAMTS13 can predict the mortality of COVID-19. In this review, we summarize the biological characteristics and interactions of NETs, VWF, and ADAMTS13, and discuss their roles in TMAs, AIS, and COVID-19. Targeting the NET-VWF axis may be a novel therapeutic strategy for inflammation-associated TMAs, AIS, and COVID-19.

Asymmetric N-aminoalkylation of 3-substituted indoles by N-protected N,O-acetals: an access to chiral propargyl aminals.

A direct enantioselective N1 aminoalkylation of 3-substituted indoles is efficiently catalyzed by a phosphoric acid catalyst under mild conditions to afford diverse enantioenriched propargyl aminals. The strategy could be applied to the modification of tryptophan containing oligopeptides. Additionally, structurally diverse and multifunctional transformations of the propargyl aminal products reveal the potential synthetic utility of this protocol.

Cancer Cell Membrane-Decorated Zeolitic-Imidazolate Frameworks Codelivering Cisplatin and Oleanolic Acid Induce Apoptosis and Reversed Multidrug Resistance on Bladder Carcinoma Cells.

Combination therapy is emerging as a preferable approach in cancer therapy with minimized side effects and elevated performance. Nevertheless, the poor targeting and drug loading of currently available drug delivery systems (DDSs) are the main difficulties to realize preferable combination therapy of cancer. As a result, a cancer cell membrane-decorated zeolitic-imidazolate framework hybrid nanoparticle (HP) was successfully constructed in our study to codeliver cisplatin (DDP) and oleanolic acid (OLA). Our results showed positive results of the platform (HP/DDP/OLA) for the treatment of bladder cancer (SW780). In detail, HP/DDP/OLA could enhance apoptosis while reverse multidrug resistance in SW780 cells than free drugs alone or monodelivery systems, which might be a suitable DDS for codelivery of different drugs with great promise.

Organocatalytic Enantioselective Synthesis of Tetrasubstituted α-Amino Allenoates by Dearomative γ-Addition of 2,3-Disubstituted Indoles to ß,γ-Alkynyl-α-imino Esters.

The first asymmetric synthesis of tetrasubstituted α-amino allenoates by a chiral phosphoric acid catalyzed dearomative γ-addition reaction of 2,3-disubstituted indoles to ß,γ-alkynyl-α-imino esters is reported. This method provides access to a series of highly functionalized tetrasubstituted allenes featuring quaternary stereocenters in high yields, and with excellent regio-, diastereo-, and enantioselectivities under mild conditions without by-product formation. Representative large-scale reactions and diverse transformations of the products into various scaffolds with potential biological activities render are also disclosed. The mechanism of the reaction was elucidated by control reactions and DFT calculations.

Characterization of adhesion properties of the cardiomyocyte integrins and extracellular matrix proteins using atomic force microscopy.

Integrins are transmembrane adhesion receptors that play important roles in the cardiovascular system by interacting with the extracellular matrix (ECM). However, direct quantitative measurements of the adhesion properties of the integrins on cardiomyocyte (CM) and their ECM ligands are lacking. In this study, we used atomic force microscopy (AFM) to quantify the adhesion force (peak force and mean force) and binding probability between CM integrins and three main heart tissue ECM proteins, ie, collagen (CN), fibronectin (FN), and laminin (LN). Functionalizing the AFM probes with ECM proteins, we found that the peak force (mean force) was 61.69 ± 5.5 pN (76.54 ± 4.0 pN), 39.26 ± 4.4 pN (59.84 ± 3.6 pN), and 108.31 ± 4.2 pN (129.63 ± 6.0 pN), respectively, for the bond of CN-integrin, FN-integrin, and LN-integrin. The binding specificity between CM integrins and ECM proteins was verified by using monoclonal antibodies, where α10 - and α11 -integrin bind to CN, α3 - and α5 -integrin bind to FN, and α3 - and α7 -integrin bind to LN. Furthermore, adhesion properties of CM integrins under physiologically high concentrations of extracellular Ca2+ and Mg2+ were tested. Additional Ca2+ reduced the adhesion mean force to 68.81 ± 4.0 pN, 49.84 ± 3.3 pN, and 119.21 ± 5.8 pN and binding probability to 0.31, 0.34, 0.40 for CN, FN, and LN, respectively, whereas Mg2+ caused very minor changes to adhesion properties of CM integrins. Thus, adhesion properties between adult murine CM integrins and its main ECM proteins were characterized, paving the way for an improved understanding of CM mechanobiology.

Access to α,γ-Diamino Diacid Derivatives via Organocatalytic Asymmetric 1,4-Addition of Azlactones and Dehydroalanines.

A convenient and functional-group-tolerant organocatalytic asymmetric 1,4-addition of azlactones and dehydroalanine is disclosed. The reaction is used for the first synthesis of chiral α,γ-diamino diacid derivatives with nonadjacent stereogenic centers in moderate to high yields, with excellent diastereo- and enantioselectivities, under the catalysis of a chiral thiourea catalyst. In addition, the reaction could be conducted in gram-scale, and the products of the reaction could be readily converted to various α,γ-diamino diacid derivatives, α,γ-diamino dialcohols, and modified peptides with nonproteinogenic amino acid residues.

Aggregation-Induced Emission Luminogen-Based Direct Visualization of Concentration Gradient Inside an Evaporating Binary Sessile Droplet.

In this study, the concentration gradient inside evaporating binary sessile droplets of 30, 50, and 60 vol % tetrahydrofuran (THF)/water mixtures was investigated. The 5 µL THF/water droplets were evaporated on a transparent hydrophobic substrate. This is the first demonstration of local concentration mapping within an evaporating binary droplet utilizing the aggregation-induced emission material. During the first two evaporation stages of the binary droplet, the local concentration can be directly visualized by the change of fluorescence emission intensity. Time-resolved average and local concentrations can be estimated by using the pre-established function of fluorescence intensity versus water volume fraction.

Endomorphin-1 analogs containing α-methyl-ß-amino acids exhibit potent analgesic activity after peripheral administration.

This study describes the design and synthesis of endomorphin-1 analogs containing C-terminal aromatic α-methyl-ß-amino acids and an N-terminal native tyrosine or 2,6-dimethyl-tyrosine. We show that, in comparison with the parent peptide, these analogs exhibit improved bioactivity and blood-brain barrier penetration after intravenous administration, and have a lower tendency to induce constipation and sedation than morphine.

Chiral Phosphoric Acid Catalyzed Asymmetric Oxidative Dearomatization of Naphthols with Quinones.

A highly enantioselective oxidative dearomatization of naphthols with quinones catalyzed by a chiral spirocyclic phosphoric acid is described. The strategy provides concise access to enantioenriched cyclohexadienones with a quinone moiety. Remarkably, the obtained products could be easily transformed to a potentially useful dihydronaphtho[2,1-b]benzofuran scaffold.

Catalyst-controlled switch of regioselectivity in the asymmetric allylic alkylation of oxazolones with MBHCs.

A catalyst-controlled switch of regioselectivity in asymmetric allylic alkylation of oxazolones with MBHCs was described. The SN2'-SN2' reaction catalysed by a quinine-derived base produced γ-selective secondary allylic oxazolone derivatives, whereas the addition-elimination reaction catalysed by an amino acid-derived bifunctional urea catalyst provided ß-selective primary adducts.