One‐Step Formulation of Protein Microparticles with Tailored Properties: Hard Templating at Soft Conditions

Formulation of therapeutic proteins into particulate forms is a main strategy for site‐specific and prolonged protein delivery as well as for protection against degradation. Precise control over protein particle size, dispersity, purity, as well as mild preparation conditions and minimal processing steps are highly desirable. It is, however, hard to fit all these criteria with conventional preparation techniques. Here a one‐step hard‐templating synthesis of microparticles composed of functional, non‐denatured protein is reported. The method is based on filling porous CaCO₃ microtemplates with the protein near to its isoelectric point (pI) followed by pH‐ or EDTA‐mediated dissolution of the tempplates. In principle, a wide variety of proteins can be converted into microparticles using this approach. The main requirement is an overlap of the protein insolubility and a template solubility for a certain parameter (here pH or EDTA). Here the formulation of insulin particles is studied in detail and it is shown that particles consisting of high molecular weight protein (catalase) can also be prepared. In this context, the synthesis of CaCO₃ templates with controlled size, the mechanism of the protein microparticle formation and mechanical properties of the microparticles are discussed. For the first time, the fabrication of mesoporous monodispersed CaCO₃ microtemplates with identical porocity but tuned diameter from 3 to 20 μm is demonstrated. The protein particle diameter can be adjusted by choosing the appropriate template size that is critical for successful pulmonary delivery of insulin. As a first step towards insulin delivery, the in vitro release of insulin at physiological conditions is studied.     

Guidelines and recommendations for indoor use of fuel cells and hydrogen systems

Hydrogen energy applications often require that systems are used indoors (e.g., industrial trucks for materials handling in a warehouse facility, fuel cells located in a room, or hydrogen stored and distributed from a gas cabinet). It may also be necessary or desirable to locate some hydrogen system components/equipment inside indoor or outdoor enclosures for security or safety reasons, to isolate them from the end-user and the public, or from weather conditions.Using of hydrogen in confined environments requires detailed assessments of hazards and associated risks, including potential risk prevention and mitigation features. The release of hydrogen can potentially lead to the accumulation of hydrogen and the formation of a flammable hydrogen-air mixture, or can result in jet-fires. Within Hyindoor European Project, carried out for the EU Fuel Cells and Hydrogen Joint Undertaking safety design guidelines and engineering tools have been developed to prevent and mitigate hazardous consequences of hydrogen release in confined environments. Three main areas are considered: Hydrogen release conditions and accumulation, vented deflagrations, jet fires and including under-ventilated flame regimes (e.g., extinguishment or oscillating flames and steady burns). Potential RCS recommendations are also identified.     

Particulate inorganic carbon production within E. huxleyi blooms in subpolar and polar seas: a satellite time series study (1998–2013)

Making use of the OC CCI (Ocean Colour Climate Change Initiative) satellite data, and based on the developed both Emiliania huxleyi bloom contouring methodologies and binary masks, areas of coccolithophore growth were distinguished without supervision from locations of mass developments of other algae. On these grounds, and employing special processing algorithms, multi-year time series of variations in occurrence, surficial extent, and content of particulate inorganic carbon (PIC) within E. huxleyi blooming areas in the North, Norwegian, Greenland, Barents, and Bering Seas were obtained for the time period 1998–2013. Analysis of algorithmic processing of the OC CCI data permitted to reveal the spatio-temporal sequence of E. huxleyi blooming events in the above seas of the North Atlantic and Arctic Oceans. During the intra-annual cycle, E. huxleyi blooms advance from temperate to higher latitudes along the pathways of the Gulf Stream propagation to the north. Annually, the blooms arise in the vicinity of the Great Britain southern extremity, and further on move northward along the western coast of this Island Country, round it to eventually appear first in the North and Norwegian Seas (in early June), then in the Greenland Sea (in late June), and finally in the Barents Sea (in late July–early September). The regularities in dates of bloom outbreak and their duration are revealed. The bloom areas in the North Atlantic–Arctic water are the lowest in the Greenland Sea (10,000–30,000 km²) and by an order of magnitude higher in the Barents Sea. The same pattern holds for the total PIC content within E. huxleyi blooms: 400 t–14 kt in the Greenland Sea and about 0.35 Mt in the Barents Sea. In the Bering Sea, the temporal and spatial dynamics of E. huxleyi development proved to be highly irregular: before and after the 1997–2001 period of high intensity of this phenomenon, the blooms are sporadic, their extent and PIC production are either very low or insignificant. Regarding the range of E. huxleyi bloom areas in the Bering Sea during 1997–2001, it is rather similar to that of the Barents Sea. However, the PIC content in the Bering Sea, as compared to that in the Barents Sea, is higher by a factor of about two with maximum values reaching 0.4 Mt and, on one occasion (in 2001), even about 0.7 Mt.     

Photonic Microhand with Autonomous Action

Grabbing and holding objects at the microscale is a complex function, even for microscopic living animals. Inspired by the hominid‐type hand, a microscopic equivalent able to catch microelements is engineered. This microhand is light sensitive and can be either remotely controlled by optical illumination or can act autonomously and grab small particles on the basis of their optical properties. Since the energy is delivered optically, without the need for wires or batteries, the artificial hand can be shrunk down to the micrometer scale. Soft material is used, in particular, a custom‐made liquid‐crystal network that is patterned by a photolithographic technique. The elastic reshaping properties of this material allow finger movement, using environmental light as the only energy source. The hand can be either controlled externally (via the light field), or else the conditions in which it autonomously grabs a particle in its vicinity can be created. This microrobot has the unique feature that it can distinguish between particles of different colors and gray levels. The realization of this autonomous hand constitutes a crucial element in the development of microscopic creatures that can perform tasks without human intervention and self‐organized automation at the micrometer scale.     

Combination of Reaction and Separation in Heterogeneous Catalytic Hydrogenation of Ethylformate

Continuous hydrogenation reaction of ethyl benzoylformate was studied over a (–)‐cinchonidine (CD)‐modified Pt/Al₂O₃ catalyst. The catalyst showed a good stability, and high enantioselectivity was achieved in the fixed‐bed reactor. Chromatographic separation of (R)‐ and (S)‐ethyl mandelate originating from a post‐continuous hydrogenation reaction of ethyl benzoylformate over the (–)‐CD‐modified Pt/Al₂O₃ catalyst was investigated in the same reaction mixture. A commercial column filled with a chiral selector resin was chosen as a perspective preparative‐scale adsorbent. Since adsorption equilibrium isotherms were linear within the entire investigated range of concentrations, they were determined by pulse experiments for the isomers present in a post‐reaction mixture. Breakthrough curves were measured and described successfully by the dispersive plug‐flow model with linear driving force approximation.     

Peculiar electric properties of polarized layer in alkaline silicate glasses

Broadband dielectric spectroscopy (BDS) was applied to study polarization phenomena in alkaline silicate glasses, in particular, properties and structure of subsurface (anodic) polarized layers forming in poling with deposited film electrodes of different structures. A model of poled glasses which does not contradict experimental data is proposed. In accordance with the model, a poled glass is presented as two resistor-capacitor circuits in a series connection, one of which is the polarized layer and another is the rest of the sample. It is found that the electric properties of the layers essentially depend on the structure of the anodic electrode used in glass poling. It is also shown that the dielectric response of poled glass samples is mainly determined by the electric properties of the submicron polarized layers and this gives an opportunity to reveal specific properties of the layers rather than ones of the glass sample bulk. Revealed temperature dependence of DC conductivity of the polarized layers obeys Arrhenius's law, and determining activation energy does not depend on the electrode. Finally, it is noted that today above-mentioned information about polarized layers can be obtained only by BDS.     

Phase-locking of multicore fibre laser due to talbot self-reproduction

Abstract

The multicore fibre laser (MCFL), containing an array of single-mode microcores in a circle inside the pump core, is a promising compact laser source. The problem is to synchronize the radiation of microcores with different propagation constants at a given radiation frequency. The theory of phase-locking of an MCFL with an external mirror and an annular waveguide matched to the multicore fibre and having a length some fraction of the Talbot distance is developed. Collective mode selection appears to occur due to spatial filtering at fractional Talbot distances, while the radiation frequency self-adjusts within the spectral gain range to minimize losses. Parallel coupling between microcores is achieved in the limit of a small fill factor. The maximum number of microcores that can be coupled is found. Good agreement is achieved between the theory and previously published experimental results.     

DNA Comet Giemsa Staining for Conventional Bright-Field Microscopy

This study was undertaken to evaluate the compatibility of Giemsa staining protocol with the comet assay. We showed, for the first time, that DNA comets can be visualized and analyzed using Giemsa staining. We generated DNA damage dose response curves for human peripheral blood lymphocytes exposed to X-ray radiation using the comet assay with either SybrGreen I or Giemsa stain. The dose response curves were fitted by linear regressions (R² > 0.977). The SybrGreen I results showed only ~1.2-fold higher slope coefficient (method sensitivity) compared to the Giemsa results. The unexpectedly high sensitivity of Giemsa staining for the comet assay is due to the Romanowsky-Giemsa effect, the stain photo-stability and the higher resolution of bright-field imaging compared to fluorescence imaging. Our results demonstrate that Giemsa staining can effectively be used for measuring DNA damage by the comet assay. The low cost and availability of Giemsa stain makes this method affordable for any low budget research and will facilitate new applications of the comet assay in biology and medicine.