The use of physical and virtual infrastructures for the validation of algal cryopreservation methods in international culture collections

Two cryopreservation methods, colligative cryoprotection coupled with controlled cooling and vitrification-based, encapsulation-dehydration were validated by five members of the EU research infrastructure consortium, COBRA, and two independent external validators. The test strain Chlorella vulgaris SAG 211-11b was successfully cryopreserved using two-step cooling employing passive (Mr Frosty) and Controlled Rate Freezers (CRF) attaining the desired recovery target within 15% of the median viability level (94%). Significant differences (p < 0.05) between cooling regimes were observed where Mr Frosty was more variable (Inter-Quartile Range being 21.5%, versus 13.0% for CRF samples). Viability assessment using fluorescein diacetate gave significantly (P < 0.0001) higher survival than growth in agar with median values being 96% and 89%, respectively. On employing encapsulation-dehydration, greater variability between some validators was observed, with six labs observing recovery in 100% of the beads (84-95% of cells surviving) and one lab observing survival in 80% of the treated beads. Bead disruption followed by algal growth in agar was considered the most reliable and accurate method of assessing cell survival for encapsulation-dehydration.     

Synthesis of benzannulated N-heterocycles by a palladium-catalyzed C-C/C-N coupling of bromoalkylamines

A palladium-catalyzed domino intermolecular alkylation/intramolecular amination of functionalized aryl iodides represents a new strategy for the synthesis of benzannulated N-heterocycles, affording functionalized indolines and tetrahydroquinolines from simple precursors.     

Analysis of first stage ignition delay times of dimethyl ether in a laminar flow reactor

The combustion chemistry of the first stage ignition and chemistry/flow interactions are studied for dimethyl ether (DME) with a mathematical analysis of two systems: a plug flow reactor study is used to reduce the reaction chemistry systematically. A skeletal reaction mechanism for the low temperature chemistry of DME until the onset of ignition is derived on the basis of the detailed DME mechanism of the Lawrence Livermore National Laboratory – see Curran, Fischer and Dryer, Int. J. Chem. Kinetics, Vol. 32 (2000). It is shown that reasonably good results for ignition delay times can be reached using a simple system of three ordinary differential equations and that the resulting analytical solution depends only on two reaction rates and the initial fuel concentration. The stepwise reduction of the system based on assumptions yields an understanding on why these reactions are so important. Furthermore, the validation of the assumptions yields insight into the influence of the fuel and the oxygen concentration on the temperature during the induction phase. To investigate the influence of chemistry/flow interactions, a 2D model with a laminar Hagen–Poiseuille flow and 2D-polynomial profiles for the radial species concentration is considered. For the 2D model, it is found that only the diffusion coefficients and the reactor radius need to be taken into consideration additionally to describe the system sufficiently. Also, the coupling of flow and chemistry is clarified in the mathematical analysis. The insight obtained from the comparison of the 2D model and the plug flow model is used to establish an average velocity for the conversion of ignition locations to ignition delay times in a laminar flow reactor. Finally, the 2D analytical solution is compared against new experimental data, obtained in such a laminar flow reactor for an undiluted DME/air mixture with an equivalence ratio of φ = 0.835 and a temperature range of 555 to 585 K at atmospheric pressure.     

Myosin-cross-reactive antigen (MCRA) protein from <Emphasis Type="Italic">Bifidobacterium breve</Emphasis> is a FAD-dependent fatty acid hydratase which has a function in stress protection

Background  

The aim of this study was to determine the catalytic activity and physiological role of myosin-cross-reactive antigen (MCRA) from Bifidobacterium breve NCIMB 702258. MCRA from B. breve NCIMB 702258 was cloned, sequenced and expressed in heterologous hosts (Lactococcus and Corynebacterium) and the recombinant proteins assessed for enzymatic activity against fatty acid substrates.     

p-tert-Butyl thiacalix[4]arenes functionalized at the lower rim by amide, hydroxyl and ester groups as anion receptors

New p-tert-butyl thiacalix[4]arenes differently substituted at the lower rim with amide, hydroxyl and ester groups were synthesized. Binding properties of the compounds toward some tetrabutylammonium salts n-Bu(4)NX (X = F(-), Cl(-), Br(-), I(-), CH(3)CO(2)(-), H(2)PO(4)(-), NO(3)(-)) were studied by UV spectroscopy. It was found that the stoichiometry of the complexes, generally, is 1 : 1, and the association constants are in the range of 10(3)-10(5) M(-1). The p-tert-butyl thiacalix[4]arenes containing secondary amide groups trisubstituted at the lower rim bind the studied anions most effectively. Selective receptors for fluoride and dihydrogen phosphate salts of tetrabutylammonium were found.     

Cytokinin and auxin interactions in plant development: metabolism, signalling, transport and gene expression

Auxin and cytokinins have been identified as key regulators of plant development. Recently, these phytohormones have been shown to interact during important developmental processes, including positioning, identity acquisition and maintenance of meristem organizing centres, regulation of balance between cell division and differentiation, and postembryonic de novo organogenesis. Here, we discuss recent advances in our understanding of the underlying molecular mechanisms at the levels of regulating metabolism, signalling, gene expression and protein stability.     

Characterization of collagen thin films for von Willebrand factor binding and platelet adhesion

Von Willebrand factor (VWF) binding and platelet adhesion to subendothelial collagens are initial events in thrombus formation at sites of vascular injury. These events are often studied in vitro using flow assays designed to mimic vascular hemodynamics. Flow assays commonly employ collagen-functionalized substrates, but a lack of standardized methods of surface ligation limits their widespread use as a clinical diagnostic. Here, we report the use of collagen thin films (CTF) in flow assays. Thin films were grown on hydrophobic substrates from type I collagen solutions of increasing concentration (10, 100, and 1000 μg/mL). We found that the corresponding increase in fiber surface area determined the amount of VWF binding and platelet adhesion. The association rate constant (k(a)) of plasma VWF binding at a wall shear stress of 45 dyn/cm(2) was 0.3 × 10(5), 1.8 × 10(5), and 1.6 × 10(5) M(-1) s(-1) for CTF grown from 10, 100, and 1000 μg/mL solutions, respectively. We observed a 5-fold increase in VWF binding capacity with each 10-fold increase in collagen solution concentration. The association rates of Ser1731Thr and His1786Asp VWF mutants with collagen binding deficiencies were 9% and 22%, respectively, of wild-type rates. Using microfluidic devices for blood flow assays, we observed that CTF supported platelet adhesion at a wall shear rate of 1000 s(-1). CTF grown from 10 and 100 μg/mL solutions had variable levels of platelet surface coverage between multiple normal donors. However, CTF substrates grown from 1000 μg/mL solutions had reproducible surface coverage levels (74 ± 17%) between normal donors, and there was significantly diminished surface coverage from two type 1 von Willebrand disease patients (8.0% and 24%). These results demonstrate that collagen thin films are homogeneous and reproducible substrates that can measure dysfunctions in VWF binding and platelet adhesion under flow in a clinical microfluidic assay format.     

A hint on the correlation between cellulose fibers polymerization degree and their thermal and thermo-oxidative degradation

The degree of polymerization is one of the main parameters reflecting cellulose ageing. Viscometry is a method frequently used for determination of cellulose fibers polymerization degree, however, sample preparation and viscosity measurement are demanding, time consuming and do not provide reproducible results. In this study, the relationship between polymerization degree of cellulose fibers obtained by viscosity measurement and their degradation parameters obtained by thermal analysis were investigated. Differential scanning calorimetry provided values of effective combustion heat and thermogravimetry (TG) was used for the determination of temperatures and associated mass losses during the cellulose degradation. Effective combustion heat did not show any correlation with degree of polymerization of investigated cotton fabrics. In contrast, results from TG suggested several promising nonlinear correlations which could be used as a hint to develop a method useful for quick determination of cotton fabrics polymerization degree. The most promising correlations with cellulose polymerization degree were found for both rates of thermal and thermo-oxidative degradations.     

Synthesis of diversely 1,3,5-trisubstituted pyrazoles via 5-exo-dig cyclization

5-Exo-dig cyclocondensation of alk-3-yn-1-ones with hydrazines, in the presence of montmorillonite K-10, provides an effective method with a high atom economy for the synthesis of diversely 1,3,5-trisubstituted pyrazoles. The microwave-accelerated reaction proceeds in the absence of solvent and leads to 5-benzyl substituted pyrazoles with good yields (72-91%). The regiochemistry of the process was confirmed by the X-ray crystallographic structure determination of 1-(2-fluorophenyl)-5-(4-methylbenzyl)-3-phenyl-1H-pyrazole.