Synthetic Modular Antibody Construction by Using the SpyTag/SpyCatcher Protein-Ligase System

Efforts to engineer recombinant antibodies for specific diagnostic and therapy applications are time consuming and expensive, as each new recombinant antibody needs to be optimized for expression, stability, bio-distribution, and pharmacokinetics. We have developed a new way to construct recombinant antibody-like “devices” by using a bottom-up approach to build them from well-behaved discrete recombinant antibody domains or “parts”. Studies on antibody structure and function have identified antibody constant and variable domains with specific functions that can be expressed in isolation. We used the SpyTag/SpyCatcher protein ligase to join these parts together, thereby creating devices with desired properties based on summed properties of parts and in configurations that cannot be obtained by using genetic engineering. This strategy will create optimized recombinant antibody devices at reduced costs and with shortened development times.     

The effect of the nature of vanadium species on benzene total oxidation

The nature of the vanadium species present on V₂O₅/Al₂O₃ catalysts was investigated by using solid state ⁵¹V NMR, diffuse reflectance spectroscopy (DRS), X-ray diffraction (XRD) and temperature programmed reduction (TPR). ⁵¹V NMR and DRS analyses indicated the presence of V⁵⁺ in tetrahedral symmetry at low vanadium loading. A surface polymeric vanadium species and/or the bulk crystalline V₂O₅ were mainly observed at high vanadium loading as also detected by XRD. The positions of the absorption edges determined through the UV–VIS spectra allowed distinguishing between various tetrahedral symmetries. After TPR, the average oxidation state of vanadium depended on the vanadium content. The nature of vanadium species was related to the catalyst behavior on the benzene oxidation reaction. The catalysts containing high vanadium content were more active suggesting that a high amount of V⁴⁺ is responsible for the higher activity.     

Modulation of human dermal microvascular endothelial cell and human gingival fibroblast behavior by micropatterned silica coating surfaces for zirconia dental implant applications

Dental ceramic implants have shown superior esthetic behavior and the absence of induced allergic disorders when compared to titanium implants. Zirconia may become a potential candidate to be used as an alternative to titanium dental implants if surface modifications are introduced. In this work, bioactive micropatterned silica coatings were produced on zirconia substrates, using a combined methodology of sol–gel processing and soft lithography. The aim of the work was to compare the in vitro behavior of human gingival fibroblasts (HGFs) and human dermal microvascular endothelial cells (HDMECs) on three types of silica-coated zirconia surfaces: flat and micropatterned (with pillars and with parallel grooves). Our results showed that cells had a higher metabolic activity (HGF, HDMEC) and increased gene expression levels of fibroblast-specific protein-1 (FSP-1) and collagen type I (COL I) on surfaces with pillars. Nevertheless, parallel grooved surfaces were able to guide cell growth. Even capillary tube-like networks of HDMEC were oriented according to the surface geometry. Zirconia and silica with different topographies have shown to be blood compatible and silica coating reduced bacteria adhesion. All together, the results indicated that microstructured bioactive coating seems to be an efficient strategy to improve soft tissue integration on zirconia implants, protecting implants from peri-implant inflammation and improving long-term implant stabilization. This new approach of micropatterned silica coating on zirconia substrates can generate promising novel dental implants, with surfaces that provide physical cues to guide cells and enhance their behavior.     

In Vitro Simultaneous Transfer of Lipids to HDL in Coronary Artery Disease and in Statin Treatment

The exchange of lipids with cells and other lipoproteins is a crucial process in HDL metabolism and for HDL antiatherogenic function. Here, we tested a practical method to quantify the simultaneous transfer to HDL of phospholipids, free-cholesterol, esterified cholesterol and triacylglycerols and to verify the lipid transfer in patients with coronary artery disease (CAD) or undergoing statin treatment. Twenty-eight control subjects without CAD, 27 with CAD and 25 CAD patients under simvastatin treatment were studied. Plasma samples were incubated with a donor nanoemulsion prepared by ultrasonication of the constituent lipids and labeled with radioactive lipids; % lipids transferred to HDL were quantified in the HDL-containing supernatant after chemical precipitation of non-HDL fractions and the nanoemulsion. The assay was precise and reproducible. Increase of temperature (4–37 °C), of incubation period (5 min to 2 h), of HDL-cholesterol concentration (33–244 mg/dL) and of mass of nanoemulsion lipids (0.075–0.3 mg/μL) resulted in increased lipid transfer from the nanoemulsion to HDL. In contrast, increasing pH (6.5–8.5) and albumin concentration (3.5–7.0 g/dL) did not affect lipid transfer. There was no difference between CAD and control non-CAD with regard to the lipid transfer, but statin treatment reduced the transfer to HDL of all four lipids. The test herein described is a valid and practical tool for exploring an important aspect of HDL metabolism.     

Enhancement of pasta antioxidant activity with oregano and carrot leaf

Five different pasta formulations were prepared using different concentrations of carrot leaf meal (CLM) and oregano leaf meal (OLM). One control formulation (wheat flour and semolina, 70:30 w/w) and four other formulations were prepared using the following amounts of leaf meals: 5% CLM and 5% OLM (I), 10% CLM (II), 10% OLM (III), and 10% CLM, 10% OLM (IV). The highest content of alpha-linolenic acid was found in formulations II and IV, which also had a reduced n-6/n-3 ratio, 3.6 (IV), when compared to the control, which was 19.8. All the samples enhanced with leaf meals showed antioxidant activity by the DPPH radical method; however, it was higher in formulation IV, which was the same observed for phenolic compounds. Pasta added with leaf meals lost more solubles, had shorter cooking time and a lower weight increase than the control pasta. All formulations had significant sensory acceptance.     

Influence of dispersant size on rheology of non-aqueous ceramic particle suspensions

The surface of alumina, which is hydrophilic in general, was made hydrophobic either by adsorption of polymer (phys-adsorption) or by an alkylation reaction with alcohol (chem-adsorption) to enable dispersion in dodecane. Hypermer A70 (8.2 nm) was used as the polymer and 1-octanol (1.2 nm), 1-decanol (1.5 nm) and 1-hexadecanol (2.5 nm) were used as the alcohol (values in brackets are the approximate thickness of the steric barrier). Rheological measurements of ceramic suspensions indicate that it is possible to achieve a high solid loading (50 vol.%) with relatively low viscosity (0.25 Pa s at 100 s^?1, the typical shear rate for pumping of liquids in pipes) as long as the stabilising molecule is large enough. The observed rheological behaviour fitted the Quemada viscosity model quite well when excluded volume effects were taken into account. Addition of 2.8 wt.% of Hypermer A70 with respect to weight of alumina was enough to stabilise the particles.     

The effect of chlorine and oxygen concentrations on the removal of mercury at an FGD-batch reactor

A series of laboratory scale experiments were conducted in an FGD-batch reactor. A synthetic flue gas was produced and directed through a CaCO₃ suspension contained in a glass reactor vessel. The suspension temperature was set at 54 °C through a water bath. In order to observe the distribution of mercury species in the system, solid, liquid and gaseous samples were taken and analysed. For gaseous mercury determination, continuous measurements were carried out, up and downstream the reactor. Furthermore, the concentration of chlorine in the scrubber solution of the system was varied from 0 to 62 g/l under different oxidative conditions.In a first approach, a concentration drop of elemental mercury coming out of the system was observed. The latter occurs only when high concentrations of Cl⁻ are present, combined with a high O₂ availability in the scrubber. It was also observed that mercury species distribution in the different phases varies, depending on the available chemical form of chlorine and oxygen concentration.     

Flavouring of imitation cheese with enzyme-modified cheeses (EMCs): Sensory impact and measurement of aroma active short chain fatty acids (SCFAs)

Enzyme-modified cheeses (EMCs) are used to impart flavour to imitation cheese products. Cheeses (pH 6 or 5.5) were formulated with 5% w/w EMC, having low, medium or high levels of lipolysis and were examined by a sensory panel. Free fatty acid analyses were performed using SPME/GC. The flavour profile of the flavoured cheeses was affected by EMC composition and pH of the cheese base. Cheeses at a pH of 6.0, flavoured with low lipolysis EMCs, were described as ‘bland’. Lowering the pH of the cheese matrix to 5.5 appeared to increase the flavour intensity of the cheese flavoured with low lipolysis EMC and panellists ranked this cheese the highest, describing its flavour as ‘well-balanced and ‘cheesy’. This study shows that the flavours of imitation cheeses are influenced by the level of lipolysis of the EMCs used to flavour them and also by the pH of the cheese base.     

Simultaneous determination of flavor enhancers in stock cube samples by using spectrophotometric data and multivariate calibration

Spectrophotometric data followed by a suitable treatment of chemometric analysis were used for the simultaneous determination of monosodium glutamate (MSG), guanosine 5′-monophosphate (GMP) and inosine 5′-monophosphate (IMP) in stock cube samples, without any previous extraction step. By this way, the overlapping of the absorption spectra was resolved using a PLS-1 model. The concentration for experimental calibration matrix were varied between 5.03–34.2 μg mL⁻¹ for IMP and GMP, and 448–1399 μg mL⁻¹ for MSG. The relative errors of prediction (REPCV %) were 1.8, 2.8 and 3.1 for IMP, GMP and MSG, respectively.     

An assessment of advanced mobile services acceptance: Contributions from TAM and diffusion theory models

Today, in addition to traditional mobile services, there are new ones already being used, thanks to the advances in 3G-related technologies. Our work contributed to the emerging body of research by integrating TAM and Diffusion Theory. Based on a sample of 542 Dutch consumers, we found that traditional antecedents of behavioral intention, ease of use and perceived usefulness, can be linked to diffusion-related variables, such as social influence and perceived benefits (flexibility and status).