Starch-based polymers have been proposed for different tissue engineering applications due to their inherent properties. In this work, a polymeric blend of starch-poly-(?-caprolactone) (SPCL) was processed using supercritical fluid technology, namely, by supercritical assisted phase inversion. As SPCL is a biodegradable polymer, the matrices produced are susceptible of undergoing enzymatic degradation upon implantation in the human body. In vitro assessment of the enzymatic degradation of SPCL was carried out in different buffer solutions containing a-amylase and/or lipase. The effect of the presence ofthese enzymes was studied by monitoring different parameters in order to characterise both bulk and the surface of the scaffolds. As regards to bulk analysis, weight loss of the samples incubated for 1, 3, 7, 14 and 21 days was determined, further differential scanning calorimetry was carried out. The morphology of the scaffolds after these periods was analysed by micro-computed tomography (?-CT) and surface chemistry was characterised by infra-red spectroscopy and contact angle measurements. Results suggest that SPLC scaffolds undergo bulk degradation, which is typically characterised by hydrolysis of chemical bonds in the polymer chain at the centre of the matrix, resulting in a highly porous material. ? 2010 Elsevier Ltd. All rights reserved.

In the tissue engineering (TE) field, the concept of producing multifunctional scaffolds, capable not only of acting as templates for cell transplantation but also of delivering bioactive agents in a controlled manner, is an emerging strategy aimed to enhance tissue regeneration. In this work, a complex hybrid release system consisting in a three-dimensional (3D) structure based on poly(d,l-lactic acid) (PDLLA) impregnated with chitosan/chondroitin sulfate nanoparticles (NPs) was developed. The scaffolds were prepared by supercritical fluid foaming at 200 bar and 35 °C, and were then characterized by scanning electron microscopy (SEM) and micro-CT. SEM also allowed to assess the distribution of the NPs within the structure, showing that the particles could be found in different areas of the scaffold, indicating a homogeneous distribution within the 3D structure. Water uptake and weight loss measurements were also carried out and the results obtained demonstrated that weight loss was not significantly enhanced although the entrapment of the NPs in the 3D structure clearly enhances the swelling of the structure. Moreover, the hybrid porous biomaterial displayed adequate mechanical properties for cell adhesion and support. The possibility of using this scaffold as a multifunctional material was further evaluated by the incorporation of a model protein, bovine serum albumin (BSA), either directly into the PDLLA foam or in the NPs that were eventually included in the scaffold. The obtained results show that it is possible to achieve different release kinetics, suggesting that this system is a promising candidate for dual protein delivery system for TE applications. © 2010 Elsevier B.V.

Time-of-flight secondary ion mass spectrometry was used to study fourhumancalculi and to compare the results with those from twelve commercially available urinary calculi minerals including three organic compounds (L-cystine, uric acid and sodium urate). Phase identification of calcium phos- phate compounds was carried out by considering the relative ion abundances of [Ca2O]R and [CaPO2]R. Deprotonated [M–H] and protonated [MRH]R uric acid were detected and used for component recognition in pure uric acid and in the mixed samples of struvite, calcium oxalate and uric acid. Iodine related to the medical history of a patient was also detected.

A laboratory-scale reactor system is built and operated to measure the kinetic of formation for single and mixed carbon dioxide-tetrahydrofuran hydrates. The T-cycle method, which is used to collect the kinetic data, is briefly discussed. For single carbon dioxide hydrate, the induction time decreases with the increase of the initial carbon dioxide pressure up to 2.96. MPa. Beyond this pressure, the induction time is becoming relatively constant with the increase of initial carbon dioxide pressure indicating that the liquid phase is completely supersaturated with carbon dioxide. Experimental results show that the inclusion of tetrahydrofuran reduces the induction time required for hydrate formation. These observations indicate hydrate nucleation process and onset growth are more readily to occur in the presence of tetrahydrofuran. In contrast, the presence of sodium chloride prolongs the induction time due to clustering of water molecules with the ions and the salting-out effects. It is also shown that the degree of subcooling required for hydrate formation is affected by the presence of tetrahydrofuran and sodium chloride in the hydrate forming system. The presence of tetrahydrofuran in the hydrate system significantly reduces the amount of carbon dioxide uptake. The apparent rate constant, k, for those systems are reported. © 2010.

This work addresses the preparation of 3D porous scaffolds of blends of chitosan and poly(l-lactic acid), CHT and PLLA, using supercritical fluid technology. Supercritical assisted phase-inversion was used to prepare scaffolds for tissue engineering purposes. The physicochemical and biological properties of chitosan make it an excellent material for the preparation of drug delivery systems and for the development of new biomedical applications in many fields from skin to bone or cartilage regeneration. On the other hand, PLLA is a synthetic biodegradable polymer widely used for biomedical applications. Supercritical assisted phase-inversion experiments were carried out in samples with different polymer ratios and different polymer solution concentrations. The effect of CHT:PLLA ratio and polymer concentration and on the morphology and topography of the scaffolds was assessed by SEM and Micro-CT. Infra-red spectroscopic imaging analysis of the scaffolds allowed a better understanding on the distribution of the two polymers within the matrix. This work demonstrates that supercritical fluid technology constitutes a new processing technology, clean and environmentally friendly for the preparation of scaffolds for tissue engineering using these materials. © 2010 Elsevier B.V.

This work demonstrates experimentally the use of mass spectrometry (MS) for on-line, quantitative monitoring of organophilic pervaporation processes operated not only under variable conditions of tem- perature of condensation/downstream pressure but also when using complex multi-component feed streams. Due to its high sensitivity, mass spectrometry is particularly suitable for on-line monitoring of aromas in dilute streams under reduced pressure, as happens with natural aromas in permeate streams. Mass spectrometry is also suitable for on-line monitoring of solvent and co-solvent permeants (water and ethanol in the selected case study), enabling for on-line determination of selectivity of the solutes of interest. A new method of calibration is proposed and validated, which correlates the intensity of the characteristic mass peak mi/z for each compound of interest with its partial pressure in the perme- ate stream. Moreover, mass spectrometry proves to be a powerful tool for studying the fractionation of aromas, recovered by integrated pervaporation–condensation processes, allowing on-line monitoring of each solute vapour in the permeate stream. Additionally, this technique enables for accurate transient studies due to the possibility of acquiring one data point every 12–15s (potentially less, if desired) in real-time.

Electrospun structures were proposed as scaffolds owing to their morphological and structural similarities with the extracellular matrix found in many native tissues. These fibrous structures were also proposed as drug release systems by exploiting the direct dependence of the release rate of a drug on the surface area. An osteogenic differentiation factor, dexamethasone (DEX), was incorporated into electrospun polycaprolactone (PCL) nanofibers at different concentrations (5, 10, 15 and 20 wt.{%} polymer), in a single-step process. The DEX incorporated into the polymeric carrier is in amorphous state, as det ermined by DSC, and does not influence the typical nanofibers morphology. In vitro drug release studies demonstrated that the dexamethasone release was sustained over a period of 15 days. The bioactivity of the released dexamethasone was assessed by cultivating human bone marrow mesenchymal stem cells (hBMSCs) on 15 wt.{%} DEX-loaded PCL NFMs, under dexamethasone-absent osteogenic differentiation medium formulation. An increased concentration of alkaline phosphatase and deposition of a mineralized matrix was observed. Phenotypic and genotypic expression of osteoblastic-specific markers corroborates the osteogenic activity of the loaded growth/differentiation factor. Overall data suggests that the electrospun biodegradable nanofibers can be used as carriers for the sustained release of growth/differentiation factors relevant for bone tissue engineering strategies. © 2010 Elsevier Ltd.

The aim of this study was to evaluate the possibility of preparing chitosan porous matrixes using supercritical fluid technology. Supercritical immersion precipitation technique was used to prepare scaffolds of a natural biocompatible polymer, chitosan, for tissue engineering purposes. The physicochemical and biological properties of chitosan make it an excellent material for the preparation of drug delivery systems and for the development of new biomedical applications in many fields from skin to bone or cartilage. Immersion precipitation experiments were carried out at different operational conditions in order to optimize the processing method. The effect of different organic solvents on the morphology of the scaffolds was assessed. Additionally, different parameters that influence the process were tested and the effect of the processing variables such as polymer concentration, temperature and pressure in the chitosan scaffold morphology, porosity and interconnectivity was evaluated by micro computed tomography. The preparation of a highly porous and interconnected structure of a natural material, chitosan, using a clean and environmentally friendly technology constitutes a new processing technology for the preparation of scaffolds for tissue engineering using these materials. © (2010) Trans Tech Publications.

In this work, a starch-based polymer, namely a blend of starch-poly(epsilon-caprolactone) was processed by supercritical assisted phase inversion process. This processing technique has been proposed for the development of 3D structures with potential applications in tissue engineering applications, as scaffolds. The use of carbon dioxide as non-solvent in the phase inversion process leads to the formation of a porous and interconnected structure, dry and free of any residual solvent. Different processing conditions such as pressure (from 80 up to 150 bar) and temperature (45 and 55 degrees C) were studied and the effect on the morphological features of the scaffolds was evaluated by scanning electron microscopy and micro-computed tomography. The mechanical properties of the SPCL scaffolds prepared were also studied. Additionally, in this work, the in vitro biological performance of the scaffolds was studied. Cell adhesion and morphology, viability and proliferation was assessed and the results suggest that the materials prepared are allow cell attachment and promote cell proliferation having thus potential to be used in some for biomedical applications.

Harold Widom proved in 1966 that the spectrum of a Toeplitz operator \(T(a)\) acting on the Hardy space \(H^p(\mathbb{T})\) over the unit circle \(\mathbb{T}\) is a connected subset of the complex plane for every bounded measurable symbol \(a\) and \(1 < p < \infty\). In 1972, Ronald Douglas established the connectedness of the essential spectrum of \(T(a)\) on \(H^2(\mathbb{T})\). We show that, as was suspected, these results remain valid in the setting of Hardy spaces \(H^p(\Gamma,w)\), \( 1 < p < \infty \), with general Muckenhoupt weights \(w\) over arbitrary Carleson curves \(\Gamma\).

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This was a cooperation with Electric Power Processing laboratory of TU Delft. During this stay organized and financed by TU Delft, S.Valtchev supported by teaching some students, PhD students and post-doc associates (in workshops?seminars).

The paramagnetic effect due to the presence of a metal center with unpaired electrons is no longer considered a hindrance in protein NMR spectroscopy. In the present work, the paramagnetic effect due to the presence of a metal center with impaired electrons was used to map the interface of an electron transfer complex. Desulfovibrio gigas cytochrome c(3) was chosen as target to study the effect of the paramagnetic probe, Fe-rubredoxin, which produced specific line broadening in the heme IV methyl resonances M2(1) and M18(1). The rubredoxin binding surface in the complex with cytochrome c(3) was identified in a heteronuclear 2D NMR titration. The identified heme methyls on cytochrome c(3) are involved in the binding interface of the complex, a result that is in agreement with the predicted complexes obtained by restrained molecular docking, which shows a cluster of possible solutions near heme IV. The use of a paramagnetic probe in (1)HNMR titration and the mapping of the complex interface, in combination with a molecular simulation algorithm proved to be a valuable strategy to study electron transfer complexes involving non-heme iron proteins and cytochromes. (C) 2009 Elsevier Inc. All rights reserved.

The isolation and characterization of a new metalloprotein containing Cu and Fe atoms is reported. The as-isolated Cu-Fe protein shows an UV-visible spectrum with absorption bands at 320 nm, 409 nm and 615 nm. Molecular mass of the native protein along with denaturating electrophoresis and mass spectrometry data show that this protein is a multimer consisting of 14 +/- 1 subunits of 15254.3 +/- 7.6 Da. Mossbauer spectroscopy data of the as-isolated Cu-Fe protein is consistent with the presence of [2Fe-2S](2+) centers. Data interpretation of the dithionite reduced protein suggest that the metallic cluster could be constituted by two ferromagnetically coupled [2Fe-2S](+) spin delocalized pairs. The biochemical properties of the Cu-Fe protein are similar to the recently reported molybdenum resistance associated protein from Desulfovibrio, D. alaskensis. Further-more, a BLAST search from the DNA deduced amino acid sequence shows that the Cu-Fe protein has homology with proteins annotated as zinc resistance associated proteins from Desulfovibrio, D. alaskensis, D. vulgaris Hildenborough, D. piger ATCC 29098. These facts suggest a possible role of the Cu-Fe protein in metal tolerance. (C) 2009 Published by Elsevier Inc.

“Theory and Practice” of TA, which is referred to in the title of this journal “TATuP”, is usually addressed as a question of TA research. But science is more than research: the field of teaching requires just as much attention, both practically and theoretically. Therefore, a mere collection of individual teaching experiences and best practice examples does not provide a strong enough basis to discuss questions of TA teaching, these must also be embedded in a theoretical context and discussed in their relation to research. In this special issue, we aim to contribute to a combination of theoretical and practical approaches to the relation of TA and “Bildung”.

The blue or Type 1 (T1) copper site of Paracoccuspantotrophus pseudoazurin exhibits significant absorption intensity in both the 450 and 600 nm regions. These are sigma and pi S(Cys) to Cu(2+) charge transfer (CT) transitions. The temperature dependent absorption, EPR, and resonance Raman (rR) vibrations enhanced by these bands indicate that a single species is present at all temperatures. This contrasts the temperature dependent behavior of the T1 center in nitrite reductase [S. Ghosh, X. Xie, A. Dey, Y. Sun, C. Scholes, E. Solomon, Proc. Natl. Acad. Sci. 106 (2009) 4969-4974] which has a thioether ligand that is unconstrained by the protein. The lack of temperature dependence in the T1 site in pseudoazurin indicates the presence of a protein constraint similar to the blue Cu site in plastocyanin where the thioether ligand is constrained at 2.8 A. However, plastocyanin exhibits only pi CT. This spectral difference between pseudoazurin and plastocyanin reflects a coupled distortion of the site where the axial thioether in pseudoazurin is also constrained, but at a shorter Cu-S(Met) bond length. This leads to an increase in the Cu(2+)-S(Cys) bond length, and the site undergoes a partial tetragonal distortion in pseudoazurin. Thus, its ground state wavefunction has both sigma and pi character in the Cu(2+)-S(Cys) bond.