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.     

Modellbasierte Entscheidungsunterstützung in Produktions- und Dienstleistungsnetzwerken

In this paper, we sketch some of the challenges that should be addressed in future research efforts for model-based decision support in manufacturing and service networks. This includes integration issues, taking into account the autonomy of the decision-making entities in face of information asymmetry, the modeling of preferences of the decision-makers, efficiently determining robust solutions, i.e. solutions that are insensitive with respect to changes in the problem data, and a reduction of the time needed for model building and usage. The problem solution cycle includes problem analysis, the design of appropriate algorithms and their performance assessment. We are interested in a prototypical integration of the proposed methods within application systems, which can be followed up with field tests of the extended application systems. We argue that the described research agenda requires the interdisciplinary collaboration of business and information systems engineering researchers with colleagues from management science, computer science, and operations research. In addition, we present some exemplifying, illustrative examples of relevant research results.     

Sulfite Action in Glycolytic Inhibition: In Vivo Real‐Time Observation by Hyperpolarized 13C NMR Spectroscopy

Detecting the molecular targets of xenobiotic substances in vivo poses a considerable analytical challenge. Here, we describe the use of an NMR‐based tracer methodology for the instantaneous in vivo observation of sulfur(IV) action on cellular metabolism. Specifically, we find that glycolytic flux is directed towards sulfite adducts of dihydroxyacetone phosphate and pyruvate as off‐pathway intermediates that obstruct glycolytic flux. In particular, the pyruvate–sulfite association hinders the formation of downstream metabolites. The apparent in vivo association constant of pyruvate and sulfite agrees with the apparent inhibition constant of CO₂ formation, thus supporting the importance of pyruvate interception in disturbing central metabolism and inhibiting NAD regeneration.     

Surface texture of the milled surface of aluminum-ceramic composite

Journal of Mechanical Science and Technology - The work concerns the assessment of the parameters of the geometric structure of the surface of an aluminum-ceramic composite after milling in various...     

Effect of AMC on fixed‐rate traffic with hard delay constraints in mobile broadband systems

The adaptive modulation and coding (AMC) technique, which has been adopted by advanced mobile telecommunication systems, supports a flexible response to the random radio behaviour. As a result, the attained transmission rate over a wireless link is time varying. Hence, resource demands are not deterministic but fluctuating even for calls with constant bit rate service requirements. Consequently, constant bit rate calls are susceptible to a forced call termination because of insufficient resources not only in a target cell during inter‐cell handoffs but also in a serving cell during radio link deterioration. Furthermore, call blocking and dropping probabilities depend on radio propagation conditions among other factors and therefore they are dissimilar throughout a service area. The latter leads to unfairness problems. We analytically measure the impact of AMC on fixed‐rate service with hard delay constraints such as voice for different signal, mobility and traffic conditions. We consider a reference case (call requests are admitted into the system provided there are enough free resources) and two classes of admission control approaches: traditional (only inter‐cell handoffs are prioritised) and modified (all ongoing calls are prioritised). The reported results reveal conditions for which AMC affects voice call performance and can serve as guidelines on admission control design. Copyright © 2013 John Wiley & Sons, Ltd.     

Characterization of CVD Bonded Tyranno® Fibers Oxidized at High Temperatures

The oxidation of highly porous ceramic matrix composites (PCMCs) based on different Tyranno^® fibers has been analyzed by means of thermogravimetry and electron microscopy. Both uncoated fibers and PCMC materials exhibit parabolic kinetics between 900°C and 1250°C, these being faster for Ti‐doped than for Zr‐doped Tyranno fibers. Oxide layers in Ti‐doped fibers are porous and partially crystalline, whereas in Zr‐doped materials a significant fraction of relatively coarse β‐SiC grains is still found embedded in the amorphous silica matrix. On the other hand, the CVD‐SiC coatings exhibit higher oxidation rates from the outer surface than from the inner one, a phenomenon that has been associated not only with the more difficult access of oxygen to the inner face but also with the highly <111>  textured structure of these coatings, for which very different oxidation rates have been published for the inward and outward directions. Cracking phenomena observed above 1100°C for long dwelling times do not lead to an acceleration of the oxidation process, which could be due to the simultaneous crystallization of the amorphous silica layers.     

Physical and Functional Properties of Blackberry Freeze- and Spray-Dried Powders

The aim of the present work was to develop two products from blackberry juice by freeze and spray drying with potential use as food colorants or healthy ingredients. A characterization of the physical and functional properties of the powdered juices was done. Maltodextrin or a mixture of trehalose and maltodextrin were assessed as carrier matrices. Freeze-dried, maltodextrin-containing powders presented the best retention of bioactive compounds and antiradical activity; however, they showed a narrow relative humidity range for storage in the glassy state. Spray-dried powders showed better physical properties, bearing higher glass transition temperature and lower molecular mobility than freeze-dried formulations.     

Design of Porous Alginate Hydrogels by Sacrificial CaCO3 Templates: Pore Formation Mechanism

Fabrication of porous alginate hydrogels with a well‐controlled architecture useful for tissue engineering is still a challenge. Here, CaCO₃‐based templating is utilized to design stable alginate gels with controlled pore dimensions in the range of 5–50 μm. The mechanism of pore formation is studied considering two factors affecting the pore size: i) osmotic pressure generated during the dissolution of sacrificial CaCO₃ templates and ii) alginate gel network density. Osmotic pressure can achieve an upper limit of 100 MPa but does not affect the gel porosity. Additional osmotic pressure (range of kPa) induced by dextrans pre‐encapsulated into CaCO₃ vaterite is also insufficient for pore enlargement. Pore stability depends merely on the gel network density and on the number of crosslinking calcium ions provided locally per unit time; pores are collapsed when template dissolution is too slow or if there is insufficient alginate concentration (below 2%). Young's modulus indicates the soft nature of the prepared hydrogels (tens of kPa) applicable as soft porous scaffolds with a tuned internal structure.     

Using ELP Repeats as a Scaffold for De Novo Construction of Gadolinium-Binding Domains within Multifunctional Recombinant Proteins for Targeted Delivery of Gadolinium to Tumour Cells

Three artificial proteins that bind the gadolinium ion (Gd³⁺) with tumour-specific ligands were de novo engineered and tested as candidate drugs for binary radiotherapy (BRT) and contrast agents for magnetic resonance imaging (MRI). Gd³⁺-binding modules were derived from calmodulin. They were joined with elastin-like polypeptide (ELP) repeats from human elastin to form the four-centre Gd³⁺-binding domain (4MBS-domain) that further was combined with F3 peptide (a ligand of nucleolin, a tumour marker) to form the F3-W4 block. The F3-W4 block was taken alone (E2-13W4 protein), as two repeats (E1-W8) and as three repeats (E1-W12). Each protein was supplemented with three copies of the RGD motif (a ligand of integrin αvβ3) and green fluorescent protein (GFP). In contrast to Magnevist (a Gd-containing contrast agent), the proteins exhibited three to four times higher accumulation in U87MG glioma and A375 melanoma cell lines than in normal fibroblasts. The proteins remained for >24 h in tumours induced by Ca755 adenocarcinoma in C57BL/6 mice. They exhibited stability towards blood proteases and only accumulated in the liver and kidney. The technological advantages of using the engineered proteins as a basis for developing efficient and non-toxic agents for early diagnosis of tumours by MRI as well as part of BRT were demonstrated.     

Effect of Bacterial Infection on Ghrelin Receptor Regulation in Periodontal Cells and Tissues

The effect of bacterial infection on the expression of growth hormone secretagogue receptor (GHS-R) was investigated in periodontal cells and tissues, and the actions of ghrelin were evaluated. GHS-R was assessed in periodontal tissues of rats with and without periodontitis. Human gingival fibroblasts (HGFs) were exposed to Fusobacterium nucleatum in the presence and absence of ghrelin. GHS-R expression was determined by real-time PCR and immunocytochemistry. Furthermore, wound healing, cell viability, proliferation, and migration were evaluated. GHS-R expression was significantly higher at periodontitis sites as compared to healthy sites in rat tissues. F. nucleatum significantly increased the GHS-R expression and protein level in HGFs. Moreover, ghrelin significantly abrogated the stimulatory effects of F. nucleatum on CCL2 and IL-6 expressions in HGFs and did not affect cell viability and proliferation significantly. Ghrelin stimulated while F. nucleatum decreased wound closure, probably due to reduced cell migration. Our results show original evidence that bacterial infection upregulates GHS-R in rat periodontal tissues and HGFs. Moreover, our study shows that ghrelin inhibited the proinflammatory actions of F. nucleatum on HGFs without interfering with cell viability and proliferation, suggesting that ghrelin and its receptor may act as a protective molecule during bacterial infection on periodontal cells.