Peptidylarginine Deiminases Post-Translationally Deiminate Prohibitin and Modulate Extracellular Vesicle Release and MicroRNAs in Glioblastoma Multiforme.

Glioblastoma multiforme (GBM) is the most aggressive form of adult primary malignant brain tumour with poor prognosis. Extracellular vesicles (EVs) are a key-mediator through which GBM cells promote a pro-oncogenic microenvironment. Peptidylarginine deiminases (PADs), which catalyze the post-translational protein deimination of target proteins, are implicated in cancer, including via EV modulation. Pan-PAD inhibitor Cl-amidine affected EV release from GBM cells, and EV related microRNA cargo, with reduced pro-oncogenic microRNA21 and increased anti-oncogenic microRNA126, also in combinatory treatment with the chemotherapeutic agent temozolomide (TMZ). The GBM cell lines under study, LN18 and LN229, differed in PAD2, PAD3 and PAD4 isozyme expression. Various cytoskeletal, nuclear and mitochondrial proteins were identified to be deiminated in GBM, including prohibitin (PHB), a key protein in mitochondrial integrity and also involved in chemo-resistance. Post-translational deimination of PHB, and PHB protein levels, were reduced after 1 h treatment with pan-PAD inhibitor Cl-amidine in GBM cells. Histone H3 deimination was also reduced following Cl-amidine treatment. Multifaceted roles for PADs on EV-mediated pathways, as well as deimination of mitochondrial, nuclear and invadopodia related proteins, highlight PADs as novel targets for modulating GBM tumour communication.

Improving polymer nanofiber quality using a modified co-axial electrospinning process.

Based on a modified coaxial electrospinning process and suitable selection of solvent mixtures as sheath fluid, a new strategy is presented for systematically improving polymer nanofiber quality. A concentric spinneret with an indented inner capillary is designed for the modified coaxial electrospinning. With a solution of 12% w/v PVP K60 in ethanol as the core electrospinning fluid, six solvents are used as sheath fluids to investigate the impact of solvent properties on the resultant PVP nanofiber quality. The PVP nanofiber quality is closely related to solvent physical-chemical properties. High quality PVP nanofibers of average diameter 130 ±10 nm with homogeneous structures and smooth surfaces are created using a solvent mixture of acetone, ethanol and DMAc in the ratio of 3:1:1(v/v/v).

Nature of the bonding in metal-silane sigma-complexes.

The nature of metal silane sigma-bond interaction has been investigated in several key systems by a range of experimental and computational techniques. The structure of [Cp'Mn(CO)(2)(eta(2)-HSiHPh(2))] 1 has been determined by single crystal neutron diffraction, and the geometry at the Si atom is shown to approximate a trigonal bipyramid; salient bond distances and angles are Mn-H(1) 1.575(14), Si-H(1) 1.806(14), Si-H(2) 1.501(13) A, and H(1)-Si-H(2) 148.5(8) degrees. This complex is similar to [Cp'Mn(CO)(2)(eta(2)-HSiFPh(2))] 2, whose structure and bonding characteristics have recently been determined by charge density studies based on high-resolution X-ray and neutron diffraction data. The geometry at the Si atom in these sigma-bond complexes is compared with that in other systems containing hypercoordinate silicon. The Mn-H distances for 1 and 2 in solution have been estimated using NMR T(1) relaxation measurements, giving a value of 1.56(3) A in each case, in excellent agreement with the distances deduced from neutron diffraction. Density functional theory calculations have been employed to explore the bonding in the Mn-H-Si unit in 1 and 2 and in the related system [Cp'Mn(CO)(2)(eta(2)-HSiCl(3))] 3. These studies support the idea that the oxidative addition of a silane ligand to a transition metal center may be described as an asymmetric process in which the Mn-H bond is formed at an early stage, while both the establishment of the Mn-Si bond and also the activation of the eta(2)-coordinated Si-H moiety are controlled by the extent of Mn --> sigma*(X-Si-H) back-donation, which increases with increasing electron-withdrawing character of the X substituent trans to the metal-coordinated Si-H bond. This delocalized molecular orbital (MO) approach is complemented and supported by combined experimental and theoretical charge density studies: the source function S(r,Omega), which provides a measure of the relative importance of each atom's contribution to the density at a specific reference point r, clearly shows that all three atoms of the Mn(eta(2)-SiH) moiety contribute to a very similar extent to the density at the Mn-Si bond critical point, in pleasing agreement with the MO model. Hence, we advance a consistent and unifying concept which accounts for the degree of Si-H activation in these silane sigma-bond complexes.

Cannabidiol Affects Extracellular Vesicle Release, miR21 and miR126, and Reduces Prohibitin Protein in Glioblastoma Multiforme Cells.

Glioblastoma multiforme (GBM) is the most common and aggressive form of primary malignant brain tumor in adults, with poor prognosis. Extracellular vesicles (EVs) are key-mediators for cellular communication through transfer of proteins and genetic material. Cancers, such as GBM, use EV release for drug-efflux, pro-oncogenic signaling, invasion and immunosuppression; thus the modulation of EV release and cargo is of considerable clinical relevance. As EV-inhibitors have been shown to increase sensitivity of cancer cells to chemotherapy, and we recently showed that cannabidiol (CBD) is such an EV-modulator, we investigated whether CBD affects EV profile in GBM cells in the presence and absence of temozolomide (TMZ). Compared to controls, CBD-treated cells released EVs containing lower levels of pro-oncogenic miR21 and increased levels of anti-oncogenic miR126; these effects were greater than with TMZ alone. In addition, prohibitin (PHB), a multifunctional protein with mitochondrial protective properties and chemoresistant functions, was reduced in GBM cells following 1 h CBD treatment. This data suggests that CBD may, via modulation of EVs and PHB, act as an adjunct to enhance treatment efficacy in GBM, supporting evidence for efficacy of cannabinoids in GBM.

Elucidation of the bonding in Mn(eta2-SiH) complexes by charge density analysis and T1 NMR measurements: asymmetric oxidative addition and anomeric effects at silicon.

The bonding in Mn(eta2-SiH) complexes is interpreted in terms of an asymmetric oxidative addition whose extent is controlled by the substitution pattern at the hypercoordinate silicon centre, and especially by the ligand trans to the eta2-coordinating SiH moiety.

5-Fluorouracil loaded Eudragit fibers prepared by electrospinning.

A series of 5-fluorouracil (5-FU) loaded core/shell electrospun fibers is reported. The fibers have shells made of Eudragit S100 (ES-100), and drug-loaded cores comprising poly(vinylpyrrolidone), ethyl cellulose, ES-100, or drug alone. Monolithic 5-FU loaded ES-100 fibers were also prepared for comparison. Electron microscopy showed all the fibers to have smooth cylindrical shapes, and clear core-shell structures were visible for all samples except the monolithic fibers. 5-FU was present in the amorphous physical form in all the materials prepared. Dissolution studies showed that the ES-100 shell was not able to prevent drug release at pH 1.0, even though the polymer is completely insoluble at this pH: around 30-80% of the maximum drug release was reached after 2h immersion at pH 1.0. These observations are ascribed to the low molecular weight of 5-FU permitting it to diffuse through pores in the ES-100 coating, and the relatively high acid solubility of the drug providing a thermodynamic impetus for this to happen. In addition, the fibers were observed to be broken or merged following 2h at pH 1.0, giving additional escape routes for the 5-FU.

Structure-tunable Janus fibers fabricated using spinnerets with varying port angles.

The preparation of Janus fibers using a new side-by-side electrospinning process is reported. By manipulating the angle between the two ports of the spinneret emitting the working fluids, Janus nanofibers with tunable structures in terms of width, interfacial area and also volume of each side can be easily fabricated.

Mebeverine-loaded electrospun nanofibers: physicochemical characterization and dissolution studies.

Both fast dissolving and sustained release drug delivery systems (DDSs) comprising mebeverine hydrochloride (MB-HCl) embedded in either povidone (PVP) K60 or Eudragit(®) L 100-55 nanofibers have been prepared by electrospinning. The fibers are found to have cylindrical morphologies with smooth surfaces, except at high drug loadings that appear to induce surface roughness (PVP) or fragmentation (Eudragit). There is a general increase in fiber diameter with drug loading. Differential scanning calorimetry and X-ray diffraction demonstrate that the drug exists in an amorphous state in the fibers. Infrared spectroscopy data indicate that the drug has good compatibility with the polymer, whereas nuclear magnetic resonance spectroscopy and high-performance liquid chromatography analyses confirmed that the MB-HCl was not degraded during the spinning process. In vitro dissolution tests of the PVP fiber mats show them to dissolve within 10 s, an improved dissolution profile over the pure drug. The Eudragit fibers show pH-dependent drug release profiles, with only very limited release at pH 2.0 but sustained release over approximately 8 h at pH 6.8. The Eudragit nanofibers have the potential to be developed as oral DDSs for localized drug release in the intestinal tract, whereas the PVP materials may find the application as buccal delivery systems or suppositories.

Fast dissolving paracetamol/caffeine nanofibers prepared by electrospinning.

A series of polyvinylpyrrolidone fibers loaded with paracetamol (PCM) and caffeine (CAF) was fabricated by electrospinning and explored as potential oral fast-dissolving films. The fibers take the form of uniform cylinders with smooth surfaces, and contain the drugs in the amorphous form. Drug-polymer intermolecular interactions were evidenced by infrared spectroscopy and molecular modeling. The properties of the fiber mats were found to be highly appropriate for the preparation of oral fast dissolving films: their thickness is around 120-130 µm, and the pH after dissolution in deionized water lies in the range of 6.7-7.2. Except at the highest drug loading, the folding endurance of the fibers was found to be >20 times. A flavoring agent can easily be incorporated into the formulation. The fiber mats are all seen to disintegrate completely within 0.5s when added to simulated saliva solution. They release their drug cargo within around 150s in a dissolution test, and to undergo much more rapid dissolution than is seen for the pure drugs. The data reported herein clearly demonstrate that electrospun PCM/CAF fibers comprise excellent candidates for oral fast-dissolving films, which could be particularly useful for children and patients with swallowing difficulties.

Pulsatile drug release from electrospun poly(ethylene oxide)-sodium alginate blend nanofibres.

Novel and highly tuneable pulsatile drug delivery systems have been prepared through the electrospinning of a blend of poly(ethylene oxide) (PEO), sodium alginate (SA), and sodium ibuprofen (SI). The resultant fibres contain crystallites of SI embedded in a PEO-SA matrix, and rather than being obtained as flat mats on the collector plate form novel three dimensional structures extending upwards the needle. Fibres were prepared with a range of loadings of SI and SA. It was found that at pH 6.8 (reminiscent of the intestinal tract) the fibres dissolve very rapidly, freeing all the embedded drug within ca. 20 minutes. However, at pH 3 (representative of the stomach pH in the fed state or in older patients) an unusual two stage release mechanism is seen. This comprises a rapid burst release, followed by a period where no further drug is released for ca. 120-150 minutes, and then a final stage of release freeing the remainder of the drug into solution. The amount of release in the initial stage, and the length of time between the first and final drug release stages, can be controlled by adjusting the SI and SA contents of the fibres respectively. This results in highly tunable pulsatile release materials.

Sensitized terbium(III) macrocyclic-phthalimide complexes as luminescent pH switches.

Four new macrocyclic-phthalimide ligands were synthesised via the coupling of N-(3-bromopropyl)phthalimide either to cyclen (1,4,7,10-tetraazacyclododecane) itself or its carboxylate-functionalized analogues, and photophysical studies were carried out on their corresponding Tb(III) complexes in aqueous media as a function of pH. Luminescence intensities of Tb·L1a­Tb·L3a were in 'switched off' mode under acidic conditions (pH < 4), and were activated on progression to basic conditions as the phthalimido functions therein were hydrolysed to their corresponding phthalamates Tb·L1b­Tb·L3b. Emission of phthalamate-based macrocyclic Tb(III) complexes Tb·L1b­Tb·L3b was in 'switched on' mode between pH 4 and 11, exhibiting high quantum yields (Φ) and long lifetimes (τ) of the order of milliseconds at pH ~ 6. Tb(III) emissions were found to decline with increasing number of chromophores. The values of Φ and τ were 46% and 2.4 ms respectively for Tb·L1b at pH ~ 6 when activated. This is the best pH-dependent sensor based on a Tb(III) complex reported to date, benefiting from the macrocyclic architecture of the ligand.

Polyacrylonitrile nanofibers coated with silver nanoparticles using a modified coaxial electrospinning process.

BACKGROUND: The objective of this investigation was to develop a new class of antibacterial material in the form of nanofibers coated with silver nanoparticles (AgNPs) using a modified coaxial electrospinning approach. Through manipulation of the distribution on the surface of nanofibers, the antibacterial effect of Ag can be improved substantially. METHODS: Using polyacrylonitrile (PAN) as the filament-forming polymer matrix, an electrospinnable PAN solution was prepared as the core fluid. A silver nitrate (AgNO3) solution was exploited as sheath fluid to carry out the modified coaxial electrospinning process under varied sheath-to-core flow rate ratios. RESULTS: Scanning electron microscopy and transmission electron microscopy demonstrated that the sheath AgNO3 solution can take a role in reducing the nanofibers' diameters significantly, a sheath-to-core flow rate ratio of 0.1 and 0.2 resulting in PAN nanofibers with diameters of 380 ± 110 nm and 230 ± 70 nm respectively. AgNPs are well distributed on the surface of PAN nanofibers. The antibacterial experiments demonstrated that these nanofibers show strong antimicrobial activities against Bacillus subtilis Wb800, and Escherichia coli dh5α. CONCLUSION: Coaxial electrospinning with AgNO3 solution as sheath fluid not only facilitates the electrospinning process, providing nanofibers with reduced diameters, but also allows functionalization of the nanofibers through coating with functional ingredients, effectively ensuring that the active antibacterial component is on the surface of the material, which leads to enhanced activity. We report an example of the systematic design, preparation, and application of a novel type of antibacterial material coated with AgNPs via a modified coaxial electrospinning methodology.

A chemically unlocked binary molecular switch.

A highly luminescent and sensitive terbium complex of a ligand comprising of a phthalimide group appended to a DO3A moiety is an active pH sensor that is conditional on its previous pH.

Electrospun nanofibers in drug delivery: recent developments and perspectives.

In this review article, some key challenges in drug delivery are first introduced and methods that have been applied in attempts to solve them enumerated. Particularly intractable problems are highlighted: these include issues of solubility, targeting and drug degradation. The technique of electrospinning is subsequently introduced, and the influence of processing parameters on the fibers produced discussed. The potential of electrospun nanofibers in drug delivery is then explored, with examples given from the recent literature to illustrate how fibers can be used to overcome hurdles in drug solubility, degradation and targeting. Future perspectives and challenges are also considered.

Preparation of core-shell PAN nanofibers encapsulated α-tocopherol acetate and ascorbic acid 2-phosphate for photoprotection.

Magnesium l-ascorbic acid 2-phosphate (MAAP) and α-tocopherol acetate (α-TAc), as the stable vitamin C and vitamin E derivative, respectively, are often applied to skin care products for reducing UV damage. The encapsulation of MAAP (0.5%, g/mL) and α-TAc (5%, g/mL) together within the polyacrylonitrile (PAN) nanofibers was demonstrated using a coaxial electrospinning technique. The structure and morphology characterizations of the core-shell fibers MAAP/α-TAc-PAN were investigated by SEM, FTIR and XRD. As a negative control, the blend nanofibers MAAP/α-TAc/PAN were prepared from a normal electrospinning method. The results from SEM indicated that the morphology and diameter of the nanofibers were influenced by concentration of spinning solution, the polymer component of the shell, the carrying agent of the core and the fabricating methods, and the core-shell nanofibers obtained at the concentration of 8% had finer and uniform structure with the average diameters of 200 ± 15nm. From in vitro release studies it could be seen that both different fiber specimens showed a gradual increase in the amount of α-TAc or MAAP released from the nanofibers. Furthermore, α-TAc and MAAP released from the blend nanofibers showed the burst release at the maximum release of ∼15% and ∼40% during the first 6h, respectively, but their release amount from the core-shell nanofibers was only 10-12% during the initial part of the process. These results showed that core-shell nanofibers alleviated the initial burst release and gave better sustainability compared to that of the blend nanofibers. The present study would provide a basis for further optimization of processing conditions to obtain desired structured core-shell nanofibers and release kinetics for practical applications in dermal tissue.

Electrospun diclofenac sodium loaded Eudragit® L 100-55 nanofibers for colon-targeted drug delivery.

Eudragit® L 100-55 nanofibers loaded with diclofenac sodium (DS) were successfully prepared using an electrospinning process, and characterized for structural and pharmacodynamic properties. The influence of solvent and drug content on fiber formation and quality was also investigated. Fiber formation was successful using a solvent mixture 5:1 (v/v) ethanol:DMAc. XRD and DSC analysis of fibers confirm electron microscopic evidence that DS is evenly distributed in the nanofibers in an amorphous state. FTIR analysis indicates hydrogen bonding occurs between the drug and the polymer, which accounts for the molecular integration of the two components. In vitro dissolution tests verified that all the drug-loaded Eudragit® L 100-55 nanofibers had pH-dependent drug release profiles, with limited, less than 3%, release at pH 1.0, but a sustained and complete release at pH 6.8. This profile of properties indicates drug-loaded Eudragit® L 100-55 nanofibers have the potential to be developed as oral colon-targeted drug delivery systems.