Synthesis of Alkyl Fructosides

The anomeric alkylation of D-fructose is efficiently catalyzed by mild acid catalysts. Degradation of fructose is negligible under the conditions applied.     

The Relation between Calcium Sequestering Capacity and Oxidation Degree of Dicarboxy-Starch and Dicarboxy-Inulin

The objective of this study was to establish the relation between the calcium sequestering capacity (SC) of dicarboxy-starch and that of dicarboxy-inulin, and the oxidation degree of these products. An S-type curve is obtained when plotting the SC of oxidized starch versus the degree of oxidation. By contrast the sequestering capacity of dicarboxy-inulin increases linearly with the degree of oxidation. The difference in behaviour of these materials can be attributed to the fact that upon starch oxidation the favourable oxydiacetate structure is obtained only after substantial conversion, whereas in inulin this structure is obtained immediately and is present in each oxidized fructose unit.     

The role of prostaglandin E2 and nitric oxide in cell death in J774 murine macrophages

We investigated the role of prostaglandin E2 (PGE2) and its interactions with nitric oxide (NO) on cell death and NO-mediated cytotoxicity in the murine macrophage cell line J774. Stimulation of the J774 cells with lipopolysaccharide together with interferon-gamma resulted in a dose-dependent cytotoxicity and production of PGE2 and NO, measured as nitrite. Our results showed a linear correlation between PGE2 release and cytotoxicity. The cyclooxygenase (COX) inhibitor indomethacin completely inhibited PGE2 biosynthesis, without affecting NO production or cell death. This supports previous reports suggesting that overproduction of endogenous PGE2 is mainly the consequence of cell death and does not cause it. In contrast, the NO synthase inhibitor N(omega)-monomethyl-L-arginine (L-NMMA) gave a significant, though incomplete suppression of NO release and cell death. This points to the presence of other cytotoxic factors besides NO. To evaluate the toxic effect solely due to NO, macrophages were exposed to the NO donor S-nitroso-N-acetyl-D,L-penicillamine (SNAP). Incubation with SNAP also resulted in a concentration-dependent cell injury and PGE2 production. When exogenously added, PGE2 protected against SNAP-mediated cytotoxicity and simultaneously increased PGE2 release into the medium, without inducing COX-2. The cytoprotection and the stimulation of PGE2 release were both reversed by indomethacin. In conclusion, PGE2 biosynthesis may represent a mechanism by which inflammatory macrophages protect themselves against the cytotoxic effects of NO.     

Wear resistance glass-ceramics with a gahnite phase obtained in CaO-MgO-ZnO-Al2O3-B2O3-SiO2 system

The thermal conditions for obtaining the glass-ceramic material of Al_0.107B_0.374Mg_0.043Zn_0.282Ca_0.100Si_0.927O₃ with a crystalline phase in the form of gahnite (ZnAl₂O₄) were specified. The activation energy E_a and the Avrami parameter n for the crystallisation process were determined with the non-isothermal DTA procedure. The maximum temperatures of crystallisation of phases, depending on the rate of heating, ranged between 800-840 °C for willemite and 870-915 °C for gahnite. The homogeneous crystalline spinel phase was obtained by heat treatment above 1000 °C. Precipitation solely of a ghanite phase from glass-ceramic causes a relative increase in its fracture toughness and wear resistance compared to the two-phase materials, i.e., K_IC = 2.12-1.65 MPam^1/2 and w_s = 0.21 × 10⁻⁴ mm³/Nm to w_s = 1.43 × 10⁻⁴ mm³/Nm.     

Activity-based profiling of retaining β-glucosidases: a comparative study

Activity-based protein profiling (ABPP) is a versatile strategy to report on enzyme activity in vitro, in situ, and in vivo. The development and use of ABPP tools and techniques has met with considerable success in monitoring physiological processes involving esterases and proteases. Activity-based profiling of glycosidases, on the other hand, has proven more difficult, and to date no broad-spectrum glycosidase activity-based probes (ABPs) have been reported. In a comparative study, we investigated both 2-deoxy-2-fluoroglycosides and cyclitol epoxides for their utility as a starting point towards retaining β-glucosidase ABP. We also investigated the merits of direct labeling and two-step bio-orthogonal labeling in reporting on glucosidase activity under various conditions. Our results demonstrate that 1) in general cyclitol epoxides are the superior glucosidase ABPs, 2) that direct labeling is the more efficient approach but it hinges on the ability of the glucosidase to be accommodated in the active site of the reporter (BODIPY) entity, and 3) that two-step bio-orthogonal labeling can be achieved on isolated enzymes but translating this protocol to cell extracts requires more investigation.     

Synthesis and characterization of CdS quantum dots with carboxylic-functionalized poly (vinyl alcohol) for bioconjugation

In the present research it is reported the synthesis and characterization of CdS nanoparticles (NPs) prepared using carboxylic-functionalized poly (vinyl alcohol) (PVA) as the ligand via aqueous route at room temperature and ambient pressure. Different molar concentrations of carboxylic-PVA and PVA were investigated aiming at producing stable colloidal systems. Carboxylic-PVA was conjugated with BSA (bovine serum albumin) and used as capping ligand in the preparation of CdS nanocrystals. UV-visible spectroscopy, photoluminescence spectroscopy, and transmission electron microscopy were used to characterize the kinetics and the relative stability of polymer-capped CdS nanocrystals. The results have clearly indicated that the carboxylic-functionalized PVA was much more effective on nucleating and stabilizing colloidal CdS nanoparticles in aqueous suspensions compared to PVA. In addition, the CdS nanocrystals were obtained in the so-called “quantum-size confinement regime”, with the calculated average size below 4.0 nm and fluorescent activity. Thus, a novel simple route was successfully developed for synthesizing nanohybrids based on quantum dots and water-soluble chemically functionalized polymers with incorporated carboxylic moiety with the possibility of direct bioconjugation.     

Coordination‐Assisted Bioorthogonal Chemistry: Orthogonal Tetrazine Ligation with Vinylboronic Acid and a Strained Alkene

Bioorthogonal chemistry can be used for the selective modification of biomolecules without interfering with any other functionality that might be present. Recent developments in the field include orthogonal bioorthogonal reactions to modify multiple biomolecules simultaneously. During our research, we observed that the reaction rates for the bioorthogonal inverse‐electron‐demand Diels–Alder (iEDDA) reactions between nonstrained vinylboronic acids (VBAs) and dipyridyl‐s‐tetrazines were exceptionally higher than those between VBAs and tetrazines bearing a methyl or phenyl substituent. As VBAs are mild Lewis acids, we hypothesised that coordination of the pyridyl nitrogen atom to the boronic acid promoted tetrazine ligation. Herein, we explore the molecular basis and scope of VBA–tetrazine ligation in more detail and benefit from its unique reactivity in the simultaneous orthogonal tetrazine labelling of two proteins modified with VBA and norbornene, a widely used strained alkene. We further show that the two orthogonal iEDDA reactions can be performed in living cells by labelling the proteasome by using a nonselective probe equipped with a VBA and a subunit‐selective VBA bearing a norbornene moiety.     

Real‐time control of a semi‐industrial fed‐batch evaporative crystallizer using different direct optimization strategies

This article presents a model‐based control approach for optimal operation of a seeded fed‐batch evaporative crystallizer. Various direct optimization strategies, namely, single shooting, multiple shooting, and simultaneous strategies, are used to examine real‐time implementation of the control approach on a semi‐industrial crystallizer. The dynamic optimizer utilizes a nonlinear moment model for on‐line computation of the optimal operating policy. An extended Luenberger‐type observer is designed to enable closed‐loop implementation of the dynamic optimizer. In addition, the observer estimates the unmeasured process variable, namely, the solute concentration, which is essential for the intended control application. The model‐based control approach aims to maximize the batch productivity, as satisfying the product quality requirements. Optimal control of crystal growth rate is the key to fulfill this objective. This is due to the close relation of the crystal growth rate to product attributes and batch productivity. The experimental results suggest that real‐time application of the control approach leads to a substantial increase, i.e., up to 30%, in the batch productivity. The reproducibility of batch runs with respect to the product crystal size distribution is achieved by thorough seeding. The simulation and experimental results indicate that the direct optimization strategies perform similarly in terms of optimal process operation. However, the single shooting strategy is computationally more expensive. © 2010 American Institute of Chemical Engineers AIChE J, 57: 1557–1569, 2011     

Application of ultrasound for start‐up of evaporative batch crystallization of ammonium sulfate in a 75‐L crystallizer

A positive effect of ultrasound on crystallization has been shown for many applications especially on small scale. Predictable scale‐up of sonocrystallization is a challenge due to the inherent dependency of ultrasound on scale. The presented research discusses the experimental application of ultrasound to induce nucleation at low supersaturation for start‐up of evaporative batch‐wise crystallization of ammonium sulfate in a 75‐L draft tube (DT) crystallizer. A comparison is made with a conventional start‐up procedure using primary nucleation or seeding. Ultrasound is applied in two geometrically different vessels of 1.2‐L connected to a 75‐L DT crystallizer. Application of ultrasound for start‐up of a 75‐L DT crystallizer shows that an optimum amount of ground seeds is better capable to suppress nucleation. A challenge for future research is to improve the efficiency of ultrasound to produce a large number of nuclei for start‐up of batch crystallization at larger scale. © 2011 American Institute of Chemical Engineers AIChE J, 2011