Glycopolymer with Sulfated Fucose and 6'-Sialyllactose as a Dual-Targeted Inhibitor on Resistant Influenza A Virus Strains.

The frequent mutations of influenza A virus (IAV) have led to an urgent need for the development of innovative antiviral drugs. Glycopolymers offer significant advantages in biomedical applications owing to their biocompatibility and structural diversity. However, the primary challenge lies in the design and synthesis of well-defined glycopolymers to precisely control their biological functionalities. In this study, functional glycopolymers with sulfated fucose and 6'-sialyllactose were successfully synthesized through ring-opening metathesis polymerization and a postmodification strategy. The optimized heteropolymer exhibited simultaneous targeting of hemagglutinin and neuraminidase on the surface of IAV, as evidenced by MU-NANA assay and hemagglutination inhibition data. Antiviral experiments demonstrated that the glycopolymer displayed broad and efficient inhibitory activity against wild-type and mutant strains of H1N1 and H3N2 subtypes in vitro, thereby establishing its potential as a dual-targeted inhibitor for combating IAV resistance.

Development of novel antivrial agents that induce the degradation of the main protease of human-infecting coronaviruses.

The family of human-infecting coronaviruses (HCoVs) poses a serious threat to global health and includes several highly pathogenic strains that cause severe respiratory illnesses. It is essential that we develop effective broad-spectrum anti-HCoV agents to prepare for future outbreaks. In this study, we used PROteolysis TArgeting Chimera (PROTAC) technology focused on degradation of the HCoV main protease (Mpro), a conserved enzyme essential for viral replication and pathogenicity. By adapting the Mpro inhibitor GC376, we produced two novel PROTACs, P2 and P3, which showed relatively broad-spectrum activity against the human-infecting CoVs HCoV-229E, HCoV-OC43, and SARS-CoV-2. The concentrations of these PROTACs that reduced virus replication by 50 % ranged from 0.71 to 4.6 µM, and neither showed cytotoxicity at 100 µM. Furthermore, mechanistic binding studies demonstrated that P2 and P3 effectively targeted HCoV-229E, HCoV-OC43, and SARS-CoV-2 by degrading Mpro within cells in vitro. This study highlights the potential of PROTAC technology in the development of broad-spectrum anti-HCoVs agents, presenting a novel approach for dealing with future viral outbreaks, particularly those stemming from CoVs.

Total synthesis and cytotoxicity evaluation of the spirostanol saponin gitonin.

The spirostanol saponin gitonin was efficiently synthesized in 12 steps (longest linear sequence) in 18.5% overall yield from the commercially available isopropyl ß-D-1-thiogalactopyranoside (IPTG) and tigogenin. A cascade two-step glycosylation and Schmidt's inverse procedure significantly facilitated the synthesis of gitonin and its derivatives. The cytotoxic activities of gitonin and its structural analogues were evaluated against A549, HepG2, and MCF-7, and most of them exhibited moderate to excellent inhibitory activity. Our study demonstrates that the removal of the ß-D-galactopyranosyl residue (attached at C-2 of the glucose unit) from gitonin would not decrease the inhibition activities; however, further cleavage of sugar units could seriously reduce the activities. A bioassay on these cancer cell lines also suggested that the presence of 2α-hydroxy on the aglycone weakened the cytotoxicity of the designed saponin.

Solution combustion synthesis of MgO-stabilized CaO sorbents using polyethylene glycol as fuel and dispersant.

The main limit for the calcium looping process is the sharp decrease of the capture capacity of the CO2 sorbents during multiple cycles. In this research, a solution combustion method was employed to synthesize MgO-stabilized CaO sorbents. Polyethylene glycol (PEG) was used as the fuel and dispersant, with the purpose to enhance the uniformity of the Ca and Mg distributions in the sorbent. The results show that highly reactive MgO-stabilized CaO sorbents can be obtained through a solution combustion method using PEG as the fuel and dispersant. The existence of MgO can effectively restrain the sintering of the sorbent, resulting in a more porous and stable micro-structure of the sorbent. The CO2 capture capacity of the MgO-stabilized CaO sorbent synthesized under the optimum conditions is 0.40 g(CO2)/g(sorbent) after 20 cycles, which is 75.3% higher than CaCO3.

Research on Monocrystalline Silicon Micro-Nano Structures Irradiated by Femtosecond Laser.

Femtosecond (fs) laser processing has received great attention for preparing novel micro-nano structures and functional materials. However, the induction mechanism of the micro-nano structures induced by fs lasers still needs to be explored. In this work, the laser-induced periodic surface structure (LIPSS) of monocrystalline silicon (Si) under fs laser irradiation is investigated. Three different layers named amorphous silicon (a-Si) layer, transition layer, and unaffected Si layer are observed after laser irradiation. The a-Si layer on the surface is generated by the resolidification of melting materials. The unaffected Si layer is not affected by laser irradiation and maintains the initial atomic structure. The transition layer consisting of a-Si and unaffected Si layers was observed under the irradiated subsurface. The phase transition mechanism of Si irradiated by fs laser is "amorphous transition", with the absence of other crystal structures. A numerical model is established to describe the fs laser-Si interaction to characterize the electronic (lattice) dynamics of the LIPSS formation. The obtained results contribute to the understanding of fs laser processing of Si at the atomic scale as well as broaden the application prospects of fs laser for treating other semiconductor materials.

Poloxamer-Based Scaffolds for Tissue Engineering Applications: A Review.

Poloxamer is a triblock copolymer with amphiphilicity and reversible thermal responsiveness and has wide application prospects in biomedical applications owing to its multifunctional properties. Poloxamer hydrogels play a crucial role in the field of tissue engineering and have been regarded as injectable scaffolds for loading cells or growth factors (GFs) in the last few years. Hydrogel micelles can maintain the integrity and stability of cells and GFs and form an appropriate vascular network at the application site, thus creating an appropriate microenvironment for cell growth, nerve growth, or bone integration. The injectability and low toxicity of poloxamer hydrogels make them a noninvasive method. In addition, they can also be good candidates for bio-inks, the raw material for three-dimensional (3D) printing. However, the potential of poloxamer hydrogels has not been fully explored owing to the complex biological challenges. In this review, the latest progress and cutting-edge research of poloxamer-based scaffolds in different fields of application such as the bone, vascular, cartilage, skin, nervous system, and organs in tissue engineering and 3D printing are reviewed, and the important roles of poloxamers in tissue engineering scaffolds are discussed in depth.

Fluoride Coatings on Magnesium Alloy Implants.

After several years of research and development, it has been reported that magnesium alloys can be used as degradable metals in some medical device applications. Over the years, fluoride coatings have received increasing research attention for improving the corrosion resistance of magnesium. In this paper, different methods for preparing fluoride coatings and the characteristics of these coatings are reported for the first time. The influence of the preparation conditions of fluoride coatings, including the magnesium substrate, voltage, and electrolyte, on the coatings is discussed. Various properties of magnesium fluoride coatings are also summarized, with an emphasis on corrosion resistance, mechanical properties, and biocompatibility. We screened experiments and papers that planned the application of magnesium fluoride coatings in living organisms. We have selected the literature with the aim of enhancing the performance of in vivo implants for reading and further detailed classification. The authors searched PubMed, SCOPUS, Web of Science, and other databases for 688 relevant papers published between 2005 and 2021, citing 105 of them. The selected time range is the last 16 years. Furthermore, this paper systematically discusses future prospects and challenges related to the application of magnesium fluoride coatings to medical products.

Zanamivir-Cholesterol Conjugate: A Long-Acting Neuraminidase Inhibitor with Potent Efficacy against Drug-Resistant Influenza Viruses.

Antiviral therapy of influenza virus infections depends heavily on two viral neuraminidase (NA) inhibitors, oseltamivir (OSV) and zanamivir (ZNV). The efficacy of OSV is challenged by the development of viral resistance, while the clinical use of ZNV is limited by its poor pharmacokinetic profile and requirement for twice-daily intranasal administration. We have developed a novel NA inhibitor by conjugating ZNV to cholesterol. The ZNV-cholesterol conjugate showed markedly improved antiviral efficacy and plasma half-life compared with ZNV. Single-dose administration of the conjugate protected the mice from lethal challenges with wild-type or mutant H1N1 influenza viruses bearing an OSV-resistant H275Y-substitution. Mechanistic studies showed that the conjugate targeted the cell membrane and entered the host cells, thereby inhibiting the NA function and the assembly of progeny virions. The ZNV-cholesterol conjugate represents a potential new treatment for influenza infections with sustained effect. Cholesterol conjugation may be an effective strategy for improving the pharmacokinetics and efficacy of other small-molecule therapeutics.

m6 A transferase KIAA1429-stabilized LINC00958 accelerates gastric cancer aerobic glycolysis through targeting GLUT1.

Emerging evidence has demonstrated that N6 -methyladenosine (m6 A) and long noncoding RNAs (lncRNAs) are both crucial regulators in gastric cancer (GC) tumorigenesis. However, the interaction of m6 A and lncRNAs in GC progression are still unclear. Here, our team discovered that lncRNA LINC00958 expression up-regulated in GC tissue and cells. Clinically, high-expression of LINC00958 was clinically correlated to lower survival of GC patients. Functionally, in vitro assays demonstrated that LINC00958 promoted the GC cells' aerobic glycolysis. Mechanistically, methylated RNA immunoprecipitation sequencing (MeRIP-Seq) found that there were m6 A-modificated sites in LINC00958, and moreover m6 A methyltransferase KIAA1429 catalyzed the m6 A modification on LINC00958 loci. Moreover, LINC00958 interacted with GLUT1 mRNA via the m6 A-dependent manner to enhance GLUT1 mRNA transcript stability, thereby positively regulating the aerobic glycolysis of GC. In conclusion, our findings reveal the function and mechanism of KIAA1429-induced LINC00958 in GC, delineating novel understanding of m6 A-lncRNA in cancer biology.

Prolonged release and shelf-life of anticoagulant sulfated polysaccharides encapsulated with ZIF-8.

Natural active polysaccharides are attracting increased attention from pharmaceutical industries for their valuable biological activities. However, the application of polysaccharides has been restricted due to their relatively large molecular weight, complex structure, and instability. Metal-organic frameworks (MOFs) have emerged to help deliver cargo to specific locations, achieving the objectives of eliminating the potential damage to the body, protecting the drugs, and improving therapeutic effectiveness. Here, a pH-responsive zeolitic imidazolate framework (ZIF-8) was synthesized to encapsulated three sulfated polysaccharides (heparin, fucan sulfate, fucosylated chondroitin sulfate) and a non-sulfated polysaccharide, hyaluronic acid. The resulting polysaccharides@ZIF-8 biocomposites showed differences in terms of morphology, particle size, encapsulation, and release efficiency. These biocomposites retained antithrombotic activity and the framework ZIF-8 effectively protected these polysaccharides from degradation and prolonged shelf-life of the anticoagulants from the unfavorable environment.

Structure-based design of 5'-substituted 1,2,3-triazolylated oseltamivir derivatives as potent influenza neuraminidase inhibitors.

Resistant viruses containing mutant neuraminidases (NAs) with diminished drug affinity continue to emerge, and new anti-influenza agents are urgently required. Several potent inhibitors targeting the hydrophobic 150-cavity of viral NAs have been developed by modifying the antiviral drugs, oseltamivir carboxylate (OSC) and zanamivir, with hydrophobic groups. Here, we describe a different strategy for exploring novel and efficient NA inhibitors by targeting the charged amino acid residues around the entrance to the 150-cavity. We synthesized a C5-substituted OSC derivative (1e) with a 4'-phenyl-1,2,3-triazolyl group capable of entering the 150-cavity, and solved the crystal structure of 1e in complex with influenza A virus N5 NA. Using the resulting structural information, we next designed and synthesized two series of OSC derivatives carrying various polar substituents at the triazolyl group of 1e and 2e, with 2e being a 5'-phenyl-1,2,3-triazole regioisomer of 1e. The NA inhibition assays demonstrated that the 2 series (2e-n) generally had superior activity compared with the 1 series (1e-n). Compound 2j, bearing a 3-phenylamino group on the triazole ring, was the most potent inhibitor of all tested NAs including an N2 NA containing the E119V OSC-resistant mutation. Moreover, 2j potently inhibited viral replication in vitro, and molecular docking studies revealed that its phenylamino group can form an additional strong hydrogen bond with residue D151 near the entrance of the 150-cavity. The design method described in this study provides useful insights into the development of novel NA inhibitors. Compound 2j warrants further structural optimization to obtain a candidate for clinical use.

Synthesis of Sialic Acids, Their Derivatives, and Analogs by Using a Whole-Cell Catalyst.

Sialic acids (Sias) are important constituents of cell surface glycans. Ready access to Sias in large quantities would facilitate the development of carbohydrate-based vaccines and small-molecule drugs. We now present a facile method for synthesizing various natural forms and non-natural derivatives or analogs of Sias by using a whole-cell catalyst, which is constructed by adding a plasmid containing necessary enzyme genes into a metabolically engineered strain of Escherichia coli. The flexible substrate tolerance of incorporated enzymes (N-acetylglucosamine 2-epimerase and N-acetylneuraminic acid aldolase) allows the cellular catalyst to convert a wide range of simple and inexpensive sugars into various Sia-related compounds through an easily scalable fermentation process. Further, syntheses using this whole-cell biotransformation in combination with three conventional enzymatic reactions provide a series of complex Sia-containing glycans (sialyloligosaccharides) and their derivatives bearing different substituents. The processes described herein should permit the large-scale and economical production of both Sias and sialyloligosaccharides, and may complement existing chemical and enzymatic strategies.

Detection and differentiation of influenza viruses with glycan-functionalized gold nanoparticles.

Accurate diagnosis of influenza viruses is difficult and generally requires a complex process because of viral diversity and rapid mutability. In this study, we report a simple and rapid strategy for the detection and differentiation of influenza viruses using glycan-functionalized gold nanoparticles (gGNPs). This method is based on the aggregation of gGNP probes on the viral surface, which is mediated by the specific binding of the virus to the glycans. Using a set of gGNPs bearing different glycan structures, fourteen influenza virus strains, including the major subtypes currently circulating in human and avian populations, were readily differentiated from each other and from a human respiratory syncytial virus in a single-step colorimetric procedure. The results presented here demonstrate the potential of this gGNP-based system in the development of convenient and portable sensors for the clinical diagnosis and surveillance of influenza viruses.

Expression and diagnostic value of CCT3 and IQGAP3 in hepatocellular carcinoma.

BACKGROUND: To evaluate plasma chaperonin containing TCP1 complex subunit 3 (CCT3) and IQ-motif-containing GTPase-activating protein-3 (IQGAP3) as biomarker for hepatocellular carcinoma (HCC) screening and diagnosis. METHODS: Blood samples were collected from 126 HCC patients with HCC, 88 patients with cirrhosis and 50 healthy volunteers to detect plasma α-fetoprotein (AFP), CCT3 and IQGAP3 levels. Plasma AFP, CCT3 and IQGAP3 protein levels were measured by enzyme linked immunosorbent assay (ELISA). RESULTS: In the plasma of HCC patients, both CCT3 and IQGAP3 were significantly higher than in patients with cirrhosis and in healthy controls (P < 0.01). CCT3 and IQGAP3 protein level correlated well with HCC etiology, tumor size, TNM stage, and child-pugh classification. CCT3 protein had higher sensitivity in the diagnosis of HCC when compared with AFP (87.3 vs 69.8 %). In addition, CCT3 and IQGAP3 protein were able to complement AFP in detecting AFP-negative HCC patients with sensitivity and specificity of 92.1 and 70.5 % and 81.6 and 71.6 %, respectively. In the small HCC group, CCT3 and IQGAP3 protein had sensitivity of 76.6 and 74.5 %, respectively. The combination of AFP + CCT3 + IQGAP3 (0.954) had significantly superior discriminative ability than AFP alone (0.815; P < 0.01). CONCLUSIONS: CCT3 and IQGAP3 are novel complementary biomarkers for HCC screening and diagnosis, especially for AFP-negative and small HCC patients.

Structure-Based Tetravalent Zanamivir with Potent Inhibitory Activity against Drug-Resistant Influenza Viruses.

Zanamivir and oseltamivir are principal influenza antiviral drugs that target viral neuraminidase (NA), but resistant viruses containing mutant NAs with diminished drug affinity are increasingly emerging. Using the structural knowledge of both drug-binding sites and their spatial arrangement on the homotetrameric NA, we have developed a tetravalent zanamivir (TZ) molecule that exhibited marked increases in NA binding affinity, inhibition of NA enzyme activity, and in vitro plus in vivo antiviral efficacy over zanamivir. TZ functioned against both human seasonal H3N2 and avian H7N9 viruses, including drug-resistant mutants. Crystal structure of a resistant N9 NA in complex with TZ explained the function, which showed that four zanamivir residues simultaneously bound to all four monomers of NA. The design method of TZ described in this study may be useful to develop drugs or ligands that target proteins with multiple binding sites. The potent anti-influenza activity of TZ makes it attractive for further development.

Enfuvirtide-PEG conjugate: A potent HIV fusion inhibitor with improved pharmacokinetic properties.

Enfuvirtide (ENF) is a clinically used peptide drug for the treatment of HIV infections, but its poor pharmacokinetic profile (T1/2 = 1.5 h in rats) and low aqueous solubility make the therapy expensive and inconvenience. In this study, we present a simple and practical strategy to address these problems by conjugating ENF with polyethylene glycol (PEG). Site-specific attachment of a 2 kDa PEG at the N-terminus of ENF resulted in an ENF-PEG (EP) conjugate with high solubility (≥3 mg/mL) and long half-life in rats (T1/2 = 16.1 h). This conjugate showed similar antiviral activity to ENF against various primary HIV-1 isolates (EC50 = 6-91 nM). Mechanistic studies suggested the sources of the antiviral potency. The conjugate bound to a functional domain of the HIV gp41 protein in a helical conformation with high affinity (Kd = 307 nM), thereby inhibiting the gp41-mediated fusion of viral and host-cell membranes. As PEG conjugation has advanced many bioactive proteins and peptides into clinical applications, the EP conjugate described here represents a potential new treatment for HIV infections that may address the unmet medical needs associated with the current ENF therapy.

Biosynthesis of the carbamoylated D-gulosamine moiety of streptothricins: involvement of a guanidino-N-glycosyltransferase and an N-acetyl-D-gulosamine deacetylase.

Streptothricins (STNs) are atypical aminoglycosides containing a rare carbamoylated D-gulosamine (D-GulN) moiety, and the antimicrobial activity of STNs has been exploited for crop protection. Herein, the biosynthetic pathway of the carbamoylated D-GulN moiety was delineated. An N-acetyl-D-galactosamine is first attached to the streptolidine lactam by the glycosyltransferse StnG and then epimerized to N-acetyl-D-gulosamine by the putative epimerase StnJ. After carbamoylation by the carbamoyltransferase StnQ, N-acetyl-D-GulN is deacetylated by StnI to furnish the carbamoylated D-GulN moiety. In vitro studies characterized two novel enzymes: StnG is an unprecedented GT-A fold N-glycosyltransferase that glycosylates the imine nitrogen atom of guanidine, and StnI is the first reported N-acetyl-D-GulN deacetylase.

Analysis of influenza virus receptor specificity using glycan-functionalized gold nanoparticles.

Recent cases of human infection with avian influenza H5N1 and H7N9 viruses underscore an urgent need for techniques that can rapidly assess their potential threat to the humans. Determination of the receptor-binding property of influenza virus is crucial to direct viral control and prevention measures. Current methods to perform this analysis are dependent on immunoanalytical strategies that use unstable biological components and complex procedures. We have developed a facile colorimetric assay to determine the interaction of the viral hemagglutinin (HA) protein with host glycan receptors using glycan-functionalized gold nanoparticles (gGNPs). This method is based on the color and absorbance changes of gold probes when the solution is simply mixed with HAs or intact viruses. The resulting sensitivity and selectivity has enabled HA/virus binding to various glycan structures to be differentiated visually and rapidly. Using this system, we have screened, in parallel, the receptor specificity of eight representative human and avian viral HAs and three whole viruses including an emerging H7N9 strain. Our results reveal the detailed receptor-binding profiles of H7N9 virus and its HA and show that they effectively bind to human-type receptors. This gGNP-based assay represents a strategy that would be helpful for developing simple and sensitive systems to probe glycan-mediated biological processes.

Inhibition of influenza virus infection with chitosan-sialyloligosaccharides ionic complex.

With the recent emergence of drug-resistant influenza viruses, effective means of preventing and treating these contagious pathogens have become imperative. The binding receptors of influenza virus are sialyloligosaccharides (SOS), which are present on the surfaces of host cells, and are therefore attractive targets for antiviral development. We report the preparation and identification of a novel influenza virus entry inhibitor, designated chitosan-SOS complex (CS complex). The CS complex was formed through noncovalent adsorption between cationic chitosan and anionic SOS, the latter derived from bovine colostrum. The preparation was accomplished in gram quantities from chitosan and bovine colostrum oligosaccharides by a one-step dialysis process. The inhibitory activity of the complex against influenza virus infection was determined by cytotoxicity inhibition assay (IC50=42 µM). This simple preparation, combined with efficient anti-infective activity and the rich natural availability of chitosan and SOS, highlights the potential of the CS complex as a safe, practical agent for influenza prevention and control.

Synthesis and assessment of globotriose-chitosan conjugate, a novel inhibitor of shiga toxins produced by Escherichia coli.

Shiga toxin (Stx)-producing Escherichia coli (STEC) causes diarrhea and colitis in humans that can develop into a life-threatening hemolytic uremic syndrome (HUS). Developing efficient means of controlling STEC diseases, for which no drugs or vaccines are currently available, remains a high priority. We report here the construction and development of chitosan conjugates bearing the Stx ligand trisaccharide globotriose to demonstrate their potential as STEC disease treatment agents. The synthesis was accomplished by grafting a globotriose derivative containing an aldehyde-functionalized aglycone to chitosan amino groups. The obtained globotriose-chitosan conjugate bound with high affinity to Stx and efficiently neutralized its toxicity on Vero cells. Moreover, Stx levels in the gut of infected mice receiving oral doses of the conjugate were greatly diminished, enabling the mice to resist a fatal STEC challenge. The conjugate appears to function as a Stx adsorbent in the gut, preventing toxin entry into the bloodstream and consequent development of HUS. As such, the conjugate could act as a novel agent against STEC disease.