Studies on the Synergistic Effect of Tandem Semi-Stable Complementary Domains on Sequence-Defined DNA Block Copolymers.

Tandem semi-stable complementary domains play an important role in life, while the role of these domains in the folding process of nucleic acid molecules has not been systematically studied. Here, we designed a clean model system by synthesizing sequence-defined DNA-OEG copolymers composed of ssDNA fragments with palindromic sequences and orthogonal oligo(tetraethylene glycol) (OEG) linkers. By altering the lengths of DNA units (6-12 nt) and OEG linkers (Xn = 0-4) separately, we systematically studied how stabilities of tandem complementary domains and connecting flexibilities affect the assembly topology. Combining experimental methods and coarse-grained molecular simulation analysis, distributions of multiple assembled conformations (mainly monomers, dimers, and clusters) were characterized. Both results indicated that tandem semi-stable complementary domains tend to form homogeneous closed circular dimers instead of larger clusters due to the synergistic enhancement effect, and the distributions of each conformation highly depend on flexibilities.

Understanding Interfacial Nanoparticle Organization through Simulation and Theory: A Review.

Understanding the behaviors of nanoparticles at interfaces is crucial not only for the design of novel nanostructured materials with superior properties but also for a better understanding of many biological systems where nanoscale objects such as drug molecules, viruses, and proteins can interact with various interfaces. Theoretical studies and tailored computer simulations offer unique approaches to investigating the evolution and formation of structures as well as to determining structure-property relationships regarding the interfacial nanostructures. In this feature article, we summarize our efforts to exploit computational approaches as well as theoretical modeling in understanding the organization of nanoscale objects at the interfaces of various systems. First, we present the latest research advances and state-of-the-art computational techniques for the simulation of nanoparticles at interfaces. Then we introduce the applications of multiscale modeling and simulation methods as well as theoretical analysis to explore the basic science and the fundamental principles in the interfacial nanoparticle organization, covering the interfaces of polymer, nanoscience, biomacromolecules, and biomembranes. Finally, we discuss future directions to signify the framework in tailoring the interfacial organization of nanoparticles based on the computational design. This feature article could promote further efforts toward fundamental research and the wide applications of theoretical approaches in designing interfacial assemblies for new types of functional nanomaterials and beyond.

De Novo Design of Entropy-Driven Polymers Resistant to Bacterial Attachment via Multicomponent Reactions.

Nonbactericidal polymers that prevent bacterial attachment are important for public health, environmental protection, and avoiding the generation of superbugs. Here, inspired by the physical bactericidal process of carbon nanotubes and graphene derivatives, we develop nonbactericidal polymers resistant to bacterial attachment by using multicomponent reactions (MCRs) to introduce molecular "needles" (rigid aliphatic chains) and molecular "razors" (multicomponent structures) into polymer side chains. Computer simulation reveals the occurrence of spontaneous entropy-driven interactions between the bacterial bilayers and the "needles" and "razors" in polymer structures and provides guidance for the optimization of this type of polymers for enhanced resistibility to bacterial attachment. The blending of the optimized polymer with commercially available polyurethane produces a film with remarkably superior stability of the resistance to bacterial adhesion after wear compared with that of commercial mobile phone shells made by the Sharklet technology. This proof-of-concept study explores entropy-driven polymers resistant to bacterial attachment via a combination of MCRs, computer simulation, and polymer chemistry, paving the way for the de novo design of nonbactericidal polymers to prevent bacterial contamination.

Solid State Characterization of Ciprofloxacin Liposome Nanocrystals.

Liposomes have been widely researched as drug delivery systems; however, the solid state form of drug inside the liposome, whether it is in solution or in a solid state, is often not studied. The solid state properties of the drug inside the liposomes are important, as they dictate the drug release behavior when the liposomes come into contact with physiological fluid. Recently, a new approach of making liposomal ciprofloxacin nanocrystals was proposed by the use of an additional freeze-thawing step in the liposomal preparation method. This paper aims to determine the solid state properties of ciprofloxacin inside the liposomes after this additional freeze-thawing cycle using cryo-TEM, small-angle X-ray scattering (SAXS), and cross-polarized light microscopy (CPLM). Ciprofloxacin precipitated in the ciprofloxacin hydrate crystal form with a unit cell dimension of 16.7 Å. The nanocrystals also showed a phase transition at 93 °C, which represents dehydration of the hydrate crystals to the anhydrate form of ciprofloxacin, verified by temperature-dependent SAXS measurements. Furthermore, the dependence of the solid state form of the nanocrystals on pH was investigated in situ, and it was shown that the liposomal ciprofloxacin nanocrystals retained their crystalline form at pH 6-10. Understanding the solid state attributes of nanocrystals inside liposomes provides improved understanding of drug dissolution and release as well as opening avenues to new applications where the nanosized crystals can provide a dissolution benefit.

Direct Comparison of Standard Transmission Electron Microscopy and Cryogenic-TEM in Imaging Nanocrystals Inside Liposomes.

The use of electron microscopy techniques in the understanding of shape and size of nanoparticles are commonly applied to drug nanotechnology, but the type of microscopy and suitability for the particles of interest can have a significant impact on the result. The size and shape of the nanoparticles are crucial in clinical applications; however, direct comparison of the results from standard transmission electron microscopy (TEM) and cryo-TEM have rarely been reported. As a useful case for comparison, liposomal drug nanocrystals are studied here. In this study, the effect of thawing temperature on the size and shape of the ciprofloxacin nanocrystals was determined. A quantitative standard TEM assay was developed to allow for high-throughput particle size analysis. These results were compared to size and shape information obtained using the cryo-TEM method. The results showed broad agreement between the two TEM methods and that ciprofloxacin nanocrystals formed shorter and thinner crystals inside the liposomes at higher thawing temperatures. The results provide confidence in the use of standard TEM to determine the size and shape distribution of solid nanoparticles (in this case, encapsulated inside liposomes) from aqueous media without fear of sample preparation altering the conclusions.

Redox- and pH-Sensitive Glycan (Polysialic Acid) Derivatives and F127 Mixed Micelles for Tumor-Targeted Drug Delivery.

With increasing application of PEGylated products, drawbacks are beginning to emerge such as the "PEG dilemma". Other promising materials may need to be found in the current situation. Endogenous polysialic acid (PSA), which is highly expressed on mammalian, bacterial, and malignant surface, may be a promising material in oncology. In this study, a dual-responsive amphiphilic PSA cholesterol derivative (PSA-CS-CH) was synthesized to explore the opportunity of PSA in targeted drug delivery systems. PSA-CS-CH, F127 mixed micelles (PF-M), and pure F127 micelles (F-M) were prepared for comparative antitumor experiments. The in vitro experiments showed that modification of PSA-CS-CH significantly increased cytotoxicity and cellular uptake. PF-M had excellent tumor microenvironment response release behavior on acidic media with high GSH levels. The in vivo fluorescence imaging and antitumor experiments showed that PF-M had excellent tumor targeting ability and great tumor suppression ability. In summary, biodegradable PSA may contribute to cancer therapy.

Cholesterol derivative-based liposomes for gemcitabine delivery: preparation, in vitro, and in vivo characterization.

As an anti-tumor drug, gemcitabine (Gem) is commonly used for the treatment of non-small cell lung cancer and pancreatic cancer. However, there are several clinical drawbacks to using Gem, including its extremely short plasma half-life and side effects. To prolong its half-life and reduce its side effects, we synthesized a derivative of Gem using cholesterol (Chol). This derivative, called gemcitabine-cholesterol (Gem-Chol), was entrapped into liposomes by a thin-film dispersion method. The particle size of the Gem-Chol liposomes was 112.57 ± 1.25 nm, the encapsulation efficiency was above 99%, and the drug loading efficiency was about 50%. In vitro studies revealed that the Gem-Chol liposomes showed delayed drug release and long-term stability at 4 °C for up to 2 months. In vivo studies also showed the superiority of the Gem-Chol liposomes, and compared with free Gem, the Gem-Chol liposomes had longer circulation time. Moreover, an anti-tumor study in H22 and S180 tumor models showed that liposomal entrapment of Gem-Chol improved the anti-tumor effect of Gem. This study reports a potential formulation of Gem for clinical application.

Analysis of a Case of Facial Nerve Injury Caused by Bee Sting in a Child.

Background: Bee sting injuries in children are accidental and occur in rural areas in summer and autumn. They have the characteristics of rapid onset, rapid change, many complications, complex treatment, and high disability rate. Patients experience various symptoms, such as vomiting, diarrhea, dyspnea, angioedema, multiple neuritis, myocardial infarction, acute renal failure, hypotension, and collapse. Systemic complications of the nervous system are rare. However, some cases of stroke, optic neuritis, and acute disseminated encephalomyelitis are related to bee stings. There are many cases of systemic multiple organ dysfunctions after bee sting injury, but there are few reports of facial nerve injury. The case presented here was caused by bee venom. This report is important because there are few instances of facial paralysis in the large number of notified bee sting cases. After active treatment, the facial paralysis of the child recovered gradually. Case Presentation: The patient was a 6-year-old boy. The bee stings by bee swarm induced pain in many parts of the body for 8 h. After the injury, he had skin itching, rash, swelling, and pain in the head and face. The boy had soy sauce-colored urine later and was transferred to the Affiliated Hospital of Zunyi Medical University from a lower-level hospital for treatment. On the seventh day after transfer, the child suddenly suffered from deviated mouth, which was considered a delayed facial nerve injury. After active treatment, he recovered from facial paralysis and was discharged from the hospital. Conclusion: This case report adds the clinical manifestation of facial paralysis after bee stings. They require close observation and being alert to possible clinical manifestations, as well as carrying out active intervention treatment.

Ultra-sensitive detection of multiplexed heavy metal ions by MOF-derived carbon film encapsulating BiCu alloy nanoparticles in potable electrochemical sensing system.

In this work, we report the development of a new type of highly active and stable Bi-based electrode material, i.e., BiCu metal-organic frames (MOF) derived carbon film (CF) encapsulating BiCu alloy nanoparticles (BiCu-ANPs) for electrochemical sensing. The integration of Bi with Cu to form BiCu-ANPs can improve their electrocatalytic activity as well as the acid resistance. Meanwhile, the carbon film that encapsulates BiCu-ANPs not only guarantees the BiCu-ANPs with high electrical conductivity and fast electrochemical kinetics but also effectively alleviates the volume change during the adsorption and desorption of heavy metal (HM) ions. Therefore, the as-obtained CF encapsulating BiCu-ANPs (BiCu-ANPs@CF) exhibits fully exposed active sites, facile charge transfer, high stability and conductivity, which gives rise to enhanced sensitivity and stability for the electrochemical detection of HM ions. When integrated into a potable electrochemical sensing system for simultaneous detection of Pb2+, Cd2+ and Zn2+, the BiCu-ANPs@CF modified electrode exhibits low detection limit (i.e., 0.081 ppb for Pb2+, 0.95 ppb for Cd2+, 35 ppb for Zn2+), wide detection range (i.e., 0.5-700 ppb for Pb2+, 5-900 ppb for Cd2+, 150-600 ppb for Zn2+) and good anti-interference. Finally, the system has been used for on-site detection of multiplexed HM ions in human biological liquids and environmental water with a good spiked recovery rate, which demanstrates its promise application in the future for on-site monitoring of human health and pollutants in water quality.

Identification and characterization of a monoclonal antibody recognizing the linear epitope RVADVI on VP1 protein of enterovirus 71.

Several large outbreaks of hand-foot-mouth disease (HFMD) have occurred in the Asian-Pacific region since 1997, with Enterovirus 71 (EV71) and/or Coxsackievirus A16 (CAV16) as the main causative agents. Despite the close genetic relationship between the two viruses, only EV71 is associated with severe clinical manifestations and deaths. Effective antiviral treatment and vaccines are not available. High-quality monoclonal antibodies (mAbs) are necessary to improve the accuracy of the diagnosis of EV71. In this study, a mAb (designated 1D9) was generated using EV71 C5 strain virus particles as immunogens. Examined by indirect immunofluorescence assay (IFA) and Western blotting, 1D9 detected successfully all 11 subgenotypes of EV71 and showed no cross-reactivity to the four selected subgenogroups of Coxsackieviruses CAV4, CAV6, CAV10, and CAV16. A linear motif, R(3) VADVI(8), which is located at the N-terminus of the EV71 VP1 protein, was identified as the minimal binding region of 1D9. Alignment and comparison of the 1D9-defined epitope sequence against the listed sequences in the NCBI EV71 database indicated that this epitope R(3) VADVI(8) was highly conserved among EV71 strains, while no significant similarity was observed when blasted against the Coxsackieviruses. This suggests that the mAb 1D9 may be useful for the development of a cost-effective and accurate method for surveillance and early differentiation of EV71 from CAV16 infection.

Development and structural characteristics of pseudoosteodentine in the Pacific cutlassfish, Trichiurus lepturus.

Depending on the mineralization pattern of dentine, teeth can be divided into three histological types (orthodont, osteodont, and pseudoosteodont type). However, the development and structural characteristics of pseudoosteodentine has not been systematically investigated yet. Here, the teeth of Trichiurus lepturus were selected for revealing a maturation process during pseudoosteodentine formation and describing ultrastructural details of pseudoosteodentine architecture. Micro-computed tomography, scanning electron microscopy, hematoxylin-eosin, Masson staining and immunohistochemistry using a dentine sialophosphoprotein (DSPP) antibody were used to analyze the microstructure and the development of the dentine. Compared with Muraenesox cinereus orthodentine, the ultrastructure of pseudoosteodentine, dentine development, the localization and migration of odontoblasts during odontogenesis in T. lepturus were observed in detail. In pseudoosteodentine, orthodentine and osteodentine all contain similar tubule-like structures and tubule openings. Labeled by DSPP immunohistochemistry for secretory odontoblasts, the organic matrix of pseudoosteodentine forms in two stages: secreting matrix by orthodentine odontoblasts with inverted nuclear polarity and the formation of osteodentine by stellate odontoblasts throughout the dental papilla. Our findings increase the understanding of the odontogenesis and structure of pseudoosteodentine and might provide a new model for the study of biomineralization and tooth development.

Entropic control of nanoparticle self-assembly through confinement.

Entropy can be the sole driving force for the construction and regulation of ordered structures of soft matter systems. Specifically, under confinement, the entropic penalty could induce enhanced entropic effects which potentially generate visually ordered structures. Therefore, spatial confinement or a crowding environment offers an important approach to control entropy effects in these systems. Here, we review how spatial confinement-mediated entropic effects accurately and even dynamically control the self-assembly of nanoscale objects into ordered structures, focusing on our efforts towards computer simulations and theoretical analysis. First, we introduce the basic principle of entropic ordering through confinement. We then introduce the applications of this concept to various systems containing nanoparticles, including polymer nanocomposites, biological macromolecular systems and macromolecular colloids. Finally, the future directions and challenges for tailoring nanoparticle organization through spatial confinement-mediated entropic effects are detailed. We expect that this review could stimulate further efforts in the fundamental research on the relationship between confinement and entropy and in the applications of this concept for designer nanomaterials.

Superstretchable, yet stiff, fatigue-resistant ligament-like elastomers.

Ligaments are flexible and stiff tissues around joints to support body movements, showing superior toughness and fatigue-resistance. Such a combination of mechanical properties is rarely seen in synthetic elastomers because stretchability, stiffness, toughness, and fatigue resistance are seemingly incompatible in materials design. Here we resolve this long-standing mismatch through a hierarchical crosslinking design. The obtained elastomer can endure 30,000% stretch and exhibit a Young's modulus of 18 MPa and toughness of 228 MJ m-3, outperforming all the reported synthetic elastomers. Furthermore, the fatigue threshold is as high as 2,682 J m-2, the same order of magnitude as the ligaments (~1,000 J m-2). We reveal that the dynamic double-crosslinking network composed of Li+-O interactions and PMMA nanoaggregates allows for a hierarchical energy dissipation, enabling the elastomers as artificial ligaments in soft robotics.

[Distribution and Sources of Microplastics in Farmland Soil Along the Fenhe River].

This study investigated the distribution and sources of microplastics smaller than 1 mm in farmland soil along the Fenhe River. Microplastics in soil samples were separated and extracted using the traditional density centrifugation method. The quantity and type of microplastics were examined with a stereomicroscope. The micro-morphology of plastic particles were observed with a scanning electron microscope-energy dispersive spectrometer. The chemical composition was determined using Fourier transform infrared spectroscopy. The results indicate that the average abundance of microplastics in farmland soil along the Fenhe River is 290.5 n ·kg-1. These microplastics occur as fibers, films, fragments, and foams. Fiber microplastics are the most abundant, accounting for 52.67% of the total, and are mostly composed of polyethylene. Films and fragments mainly consist of polypropylene whereas the foams consist of polystyrene. Soil samples from different parts of the Fenhe River can be ranked according to the microplastics content in the following order:downstream>midstream>upstream. The abundance of microplastics in soil from the downstream region of the Fenhe River was 500.0 n ·kg-1, twice that of from the upstream and midstream regions. The results of the random forest model indicate that the sources of microplastics in farmland soil along the Fenhe River are closely related to the amount of agricultural films, population, gross domestic product, and industrial production. Among these factors, the amount of agricultural films is a key factor that influences the occurrence of microplastics in farmland soil along the Fenhe River.

Enhanced Heterogeneous Diffusion of Nanoparticles in Semiflexible Networks.

The transport of nanoparticles in semiflexible networks, which form diverse principal structural components throughout living systems, is important in biology and biomedical applications. By combining large-scale molecular simulations as well as theoretical analysis, we demonstrate here that nanoparticles in polymer networks with semiflexible strands possess enhanced heterogeneous diffusion characterized by more evident hopping dynamics. Particularly, the hopping energy barrier approximates to linear dependence on confinement parameters in the regime of moderate rigidity, in contrast to the quadratic dependence of both its soft and hard counterparts. This nonmonotonic feature can be attributed to the competition between the conformation entropy and the bending energy regulated by the chain rigidity, captured by developing an analytical model of a hopping energy barrier. Moreover, these theoretical results agree reasonably well with previous experiments. The findings bear significance in unraveling the fundamental physics of substance transport confined in network-topological environments and would provide an explanation for the dynamics diversity of nanoparticles within various networks, biological or synthetic.

Exposure of liposomes containing nanocrystallised ciprofloxacin to digestive media induces solid-state transformation and altered in vitro drug release.

A recently reported approach to nanocrystallise encapsulated ciprofloxacin within liposomes has generated increased interest in the solid-state properties of drug nanocrystals within liposomal confinement. To explore the potential application of nanocrystallised drug liposomes in oral delivery, a liposomal ciprofloxacin formulation was used as a model system. An in vitro digestion model coupled to small angle X-ray scattering was used to analyse the solid-state properties of the drug nanocrystals during digestion of the liposomal ciprofloxacin nanocrystal formulations. Results showed a complete polymorphic transformation of the ciprofloxacin hydrate nanocrystals to a new salt form at a threshold sodium taurodeoxycholate to ciprofloxacin molar ratio of 0.6. The in vitro drug release from the nanocrystallised drug containing liposomes showed controlled drug release behaviour under non-digestive conditions, while a 3.5-fold increase in the drug release was seen when they were exposed to the simulated digestive environment. In conclusion, the solid state of the drug inside the liposomes is important in dictating the drug release behaviour from the liposomes. The identification of the solid state transformation during digestion in real time and the bile salt-induced polymorphic transformation of ciprofloxacin from nanocrystallised ciprofloxacin liposome are important to understand how the drug is released in vivo, as well as for future formulation design.

Distinct Interaction of Lytic Polysaccharide Monooxygenase with Cellulose Revealed by Computational and Biochemical Studies.

A distinct interaction pattern of lytic polysaccharide monooxygenases (LPMOs) with their insoluble substrate, cellulose, was revealed through the combination of computational and biochemical approaches. The results indicated that the enzymes can stably bind on the flat hydrophobic surface of cellulose via the interactions of the key residues located in the axis across the conserved distal tyrosine residue and copper ion with two adjacent cellulose chains. Further studies on the correlation of substrate binding and H2O2 accumulation suggested that LPMOs involved in the productive binding on the insoluble polysaccharides not only fail to accumulate H2O2 but also consume the H2O2 produced by the unbound molecules under the lab condition. This was further substantiated by quantum-mechanical calculations. These findings broadened our knowledge of the interaction between enzymes and insoluble substrates and deepened our understanding of the role that H2O2 plays in LPMO activity.

Controlling the size and shape of liposomal ciprofloxacin nanocrystals by varying the lipid bilayer composition and drug to lipid ratio.

Drug nanocrystals precipitated inside liposomes are of increasing interest in liposomal drug delivery. For liposomal nanocrystal formulations, the size and shape of the drug nanocrystals can influence the apparent drug release properties, providing opportunities for developing tailored liposomal drug release systems. Small angle X-ray scattering (SAXS) and quantitative transmission electron microscopy (TEM) can be used to analyse the size distributions of the nanoparticles. In this study, by changing the fluidity of the membrane through the use of different membrane phospholipids with varying cholesterol content, the impact of lipid phase, fluidity and permeability on the size distribution of ciprofloxacin nanocrystals were investigated using standard TEM and SAXS as orthogonal techniques. The results show that the phospholipid phase behaviour has a direct effect on the nanocrystal size distribution, where shorter and thinner nanocrystals were formed in liposomes made from hydrogenated soy phosphatidylcholine (HSPC) and 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) phospholipids with higher phase transition temperatures than 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) and 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) with lower transition temperatures. This is mainly due to the phase behaviour of the liposome during nanocrystal formation. The addition of cholesterol that reduces fluidity and permeability of the DOPC liposomes was also shown to restrict the growth of the ciprofloxacin nanocrystals. Moreover, increasing the drug loading of the liposomes made from HSPC and DPPC produced longer and wider nanocrystals. The findings open new opportunities to tailor nanocrystal size distributions, as well as the aspect ratio of the enclosing liposomes with potential to alter drug release and in vivo behaviour.

A lytic polysaccharide monooxygenase from Myceliophthora thermophila and its synergism with cellobiohydrolases in cellulose hydrolysis.

Lytic polysaccharide monooxygenases (LPMOs) have attracted vast attention because of their unique mechanism of oxidative degradation of carbohydrate polymers and the potential application in biorefineries. This study characterized a novel LPMO from Myceliophthora thermophila, denoted MtLPMO9L. The structure model of the enzyme indicated that it belongs to the C1-oxidizing LPMO, which has neither an extra helix in the L3 loop nor extra loop region in the L2 loop. This was confirmed subsequently by the enzymatic assays since MtLPMO9L only acts on cellulose and generates C1-oxidized cello-oligosaccharides. Moreover, synergetic experiments showed that MtLPMO9L significantly improves the efficiency of cellobiohydrolase (CBH) II. In contrast, the inhibitory rather than synergetic effect was observed when combining used MtLPMO9L and CBHI. Changing the incubation time and concentration ratio of MtLPMO9L and CBHI could attenuate the inhibitory effects. This discovery suggests a different synergy detail between MtLPMO9L and two CBHs, which implies that the composition of cellulase cocktails may need reconsideration.

Paeoniflorin Prevents Intestinal Barrier Disruption and Inhibits Lipopolysaccharide (LPS)-Induced Inflammation in Caco-2 Cell Monolayers.

Inflammatory bowel disease (IBD) in humans is closely related to bacterial infection and the disruption of the intestinal barrier. Paeoniflorin (PF), a bioactive compound from Paeonia lactiflora Pallas plants, exerts a potential effect of anti-inflammatory reported in various researches. However, the effect of PF on intestinal barrier function and its related mechanisms has not been identified. Here, we investigate the PF potential anti-inflammatory effect on lipopolysaccharide (LPS)-stimulated human Caco-2 cell monolayers and explore its underlying key molecular mechanism. In this context, PF significantly increased TEER value, decreased intestinal epithelium FITC-dextran flux permeability, and restored the expressions of occludin, ZO-1, and claudin5 in LPS-induced Caco-2 cell. In vitro, treatment of PF significantly inhibited LPS-induced expression of cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6), and matrix metalloproteinase-9 (MMP-9). In addition, we found that PF suppressed nuclear factor kappa B (NF-κB) signaling via activating the Nrf2/HO-1 signaling pathways in ILPS-stimulated Caco-2 cells. Our findings indicate that PF has an inhibitory effect on endothelial injury. Our findings suggested that PF has an anti-inflammatory effect in ILPS-stimulated Caco-2 cells, which might be a potential therapeutic agent against IBD and intestinal inflammation.