Cell-Derived Vesicles with Increased Stability and On-Demand Functionality by Equipping Their Membrane with a Cross-Linkable Copolymer

Cell-derived vesicles retain the cytoplasm and much of the native cell membrane composition. Therefore, they are attractive for investigations of membrane biophysics, drug delivery systems, and complex molecular factories. However, their fragility and aggregation limit their applications. Here, the mechanical properties and stability of giant plasma membrane vesicles (GPMVs) are enhanced by decorating them with a specifically designed diblock copolymer, cholesteryl-poly[2-aminoethyl methacrylate- b -poly(ethylene glycol) methyl ether acrylate]. When cross-linked, this polymer brush enhances the stability of the GPMVs. Furthermore, the pH-responsiveness of the copolymer layer allows for a controlled cargo loading/release, which may enable various bioapplications. Importantly, the cross-linked-copolymer GPMVs are not cytotoxic and preserve in vitro membrane integrity and functionality. This effective strategy to equip the cell-derived vesicles with stimuli-responsive cross-linkable copolymers is expected to open a new route to the stabilization of natural membrane systems and overcome barriers to biomedical applications

Mucosal and Systemic Immune Responses to Influenza H7N9 Antigen HA1–2 Co-Delivered Intranasally with Flagellin or Polyethyleneimine in Mice and Chickens

Consecutive cases of human infection with H7N9 influenza viruses since 2013 in China have prompted efforts to develop an effective treatment. Subunit vaccines introduced by intranasal administration can block an infection at its primary site; flagellin (fliC) and polyethyleneimine (PEI) have been shown to be potent adjuvants. We previously generated the hemagglutinin (HA)1–2-fliC fusion protein consisting of the globular head domain (HA1–2; amino acids 62–284) of HA fused with Salmonella typhimurium fliC. In the present study, we investigated its effectiveness of both flagellin and PEI as mucosal adjuvants for the H7N9 influenza subunit vaccine. Mice immunized intranasally with HA1–2-fliC and HA1–2-PEI showed higher HA1–2-specific immunoglobulin (Ig)G and IgA titers in serum, nasal wash, and bronchial alveolar lavage fluid. Moreover, splenocyte activation and proliferation and the number of HA1–2-specific interferon (IFN)-γ- and interleukin (IL)-4-producing splenocytes were markedly increased in the fliC and PEI groups; in the latter, there were more cells secreting IL-4 than IFN-γ, suggesting that fliC induced T helper type (Th)1 and Th2 immune responses, and PEI induced Th2-biased responses, consistent with the serum antibody isotype pattern (IgG1/IgG2a ratio). Furthermore, virus challenge was performed in a chicken model. The results showed that chickens receiving fliC and PEI adjuvant vaccine exhibited robust immune responses leading to a significant reduction in viral loads of throat and cloaca compared to chickens receiving only HA1–2. These findings provide a basis for the development of H7N9 influenza HA1–2 mucosal subunit vaccines

Association of lipoprotein(a) and major adverse cardiovascular events in patients with percutaneous coronary intervention.

Introduction(#br)The aim of the current study was to evaluate the association between lipoprotein(a) [Lp(a)] and major adverse cardiovascular events (MACEs) in patients with percutaneous coronary intervention (PCI) treatment.(#br)Material and methods(#br)This was a retrospective study. The demographics, prior medical histories, comorbidities and laboratory parameters were collected from the electronic health record. All participants were followed up for 1 year after the indexed PCI. Studied end points were a composite of MACEs including all-cause mortality, non-fatal myocardial infarction (MI), non-fatal ischemic stroke, transient ischemic attack and stent restenosis.(#br)Results(#br)During 1-year follow-up, 87 MACEs occurred. Compared to patients who did not have MACEs, patients who had MACEs were older, more likely to have higher body mass index, diabetes mellitus and left main lesion, and also had higher baseline low density lipoprotein cholesterol (LDL-C) and Lp(a) levels. All patients in both groups were prescribed aspirin and clopidogrel at discharge. Nearly 97.4% and 95.4% of patients in both groups were treated with statins and a higher proportion of patients in the MACE group were treated with ezetimibe (11.5% vs. 3.5%, p < 0.05). In multivariate regression analysis, diabetes mellitus, LDL-C, Lp(a) and glomerular filtration rate were independent risk factors for MACEs; statin use appeared to be a protective factor for MACEs. Patients with increased Lp(a) level had significantly higher incidence of MACEs than the normal Lp(a) level group ( p = 0.001).(#br)Conclusions(#br)Baseline serum Lp(a) can be used to predict MACEs in patients after PCI treatment, which was independent of LDL-C

Network Modeling Identifies Molecular Functions Targeted by miR-204 to Suppress Head and Neck Tumor Metastasis

Due to the large number of putative microRNA gene targets predicted by sequence-alignment databases and the relative low accuracy of such predictions which are conducted independently of biological context by design, systematic experimental identification and validation of every functional microRNA target is currently challenging. Consequently, biological studies have yet to identify, on a genome scale, key regulatory networks perturbed by altered microRNA functions in the context of cancer. In this report, we demonstrate for the first time how phenotypic knowledge of inheritable cancer traits and of risk factor loci can be utilized jointly with gene expression analysis to efficiently prioritize deregulated microRNAs for biological characterization. Using this approach we characterize miR-204 as a tumor suppressor microRNA and uncover previously unknown connections between microRNA regulation, network topology, and expression dynamics. Specifically, we validate 18 gene targets of miR-204 that show elevated mRNA expression and are enriched in biological processes associated with tumor progression in squamous cell carcinoma of the head and neck (HNSCC). We further demonstrate the enrichment of bottleneckness, a key molecular network topology, among miR-204 gene targets. Restoration of miR-204 function in HNSCC cell lines inhibits the expression of its functionally related gene targets, leads to the reduced adhesion, migration and invasion in vitro and attenuates experimental lung metastasis in vivo. As importantly, our investigation also provides experimental evidence linking the function of microRNAs that are located in the cancer-associated genomic regions (CAGRs) to the observed predisposition to human cancers. Specifically, we show miR-204 may serve as a tumor suppressor gene at the 9q21.1–22.3 CAGR locus, a well established risk factor locus in head and neck cancers for which tumor suppressor genes have not been identified. This new strategy that integrates expression profiling, genetics and novel computational biology approaches provides for improved efficiency in characterization and modeling of microRNA functions in cancer as compared to the state of art and is applicable to the investigation of microRNA functions in other biological processes and diseases.</p

Iterative solution to singular nth-order nonlocal boundary value problems

By using the cone theory and the Banach contraction mapping principle, we study the existence and uniqueness of an iterative solution to the singular nth-order nonlocal boundary value problems

Interhemispheric transport of metallic ions within ionospheric sporadic E layers by the lower thermospheric meridional circulation

Long-lived metallic ions in the Earth’s atmosphere/ionosphere have been investigated for many decades. Although the seasonal variation in ionospheric ‘sporadic E’ layers was first observed in the 1960s, the mechanism driving the variation remains a long-standing mystery. Here we report a study of ionospheric irregularities using scintillation data from COSMIC satellites and identify a large-scale horizontal transport of long-lived metallic ions, combined with the simulations of the Whole Atmosphere Community Climate Model with the chemistry of metals and ground-based observations from two meridional chains of stations from 1975–2016. We find that the lower thermospheric meridional circulation influences the meridional transport and seasonal variations of metallic ions within sporadic E layers. The winter-to-summer, meridional velocity of ions is estimated to vary between -1.08 and 7.45 m/s at altitudes of 107–118 km between 10�–60�N latitude. Our results not only provide strong support for the lower thermospheric meridional circulation predicted by a whole atmosphere chemistry-climate model, but also emphasise the influences of this winter-to-summer circulation on the large-scale interhemispheric transport of composition in the thermosphere/ionosphere

Two Neuronal Nicotinic Acetylcholine Receptors, α4β4 and α7, Show Differential Agonist Binding Modes

Nicotinic acetylcholine receptors (nAChRs) are pentameric, neurotransmitter-gated ion channels responsible for rapid excitatory neurotransmission in the central and peripheral nervous systems, resulting in skeletal muscle tone and various cognitive effects in the brain. These complex proteins are activated by the endogenous neurotransmitter ACh as well as by nicotine and structurally related agonists. Activation and modulation of nAChRs has been implicated in the pathology of multiple neurological disorders, and as such, these proteins are established therapeutic targets. Here we use unnatural amino acid mutagenesis to examine the ligand binding mechanisms of two homologous neuronal nAChRs: the α4β4 and α7 receptors. Despite sequence identity among the residues that form the core of the agonist-binding site, we find that the α4β4 and α7 nAChRs employ different agonist-receptor binding interactions in this region. The α4β4 receptor utilizes a strong cation-π interaction to a conserved tryptophan (TrpB) of the receptor for both ACh and nicotine, and nicotine participates in a strong hydrogen bond with a backbone carbonyl contributed by TrpB. Interestingly, we find that the α7 receptor also employs a cation-π interaction for ligand recognition, but the site has moved to a different aromatic amino acid of the agonist-binding site depending on the agonist. ACh participates in a cation-π interaction with TyrA, whereas epibatidine participates in a cation-π interaction with TyrC2