A standardized and biocompatible preparation of aggregate-free amyloid beta peptide for biophysical and biological studies of Alzheimer's disease

We provide a validated and rapid protocol for the solubilization of amyloid β-peptide (Aβ). This procedure involves sequential solubilization using structure-breaking organic solvents hexafluoroisopropanol and DMSO followed by column purification. The low solubility and tendency of Aβ to aggregate considerably impede the in vitro handling and biophysical or biological investigation of Aβ, despite the interest in this peptide because of its implication in Alzheimer's disease. The main advantage of the proposed protocol over others is that it results in standardized aggregate-free Aβ peptide samples that are biocompatible for cell culture studies and yield reproducible aggregation kinetics and cytotoxicities. This three-step protocol also enables the co-solubilization of the longer Aβ42 variant with Aβ40 in ratios relevant to Alzheimer's disease.     

The Ketosynthase Domain Controls Chain Length in Mushroom Oligocyclic Polyketide Synthases

The nonreducing iterative type I polyketide synthases (NR-PKSs) CoPKS1 and CoPKS4 of the webcap mushroom Cortinarius odorifer share 88 % identical amino acids. CoPKS1 almost exclusively produces a tricyclic octaketide product, atrochrysone carboxylic acid, whereas CoPKS4 shows simultaneous hepta- and octaketide synthase activity and also produces the bicyclic heptaketide 6-hydroxymusizin. To identify the region(s) controlling chain length, four chimeric enzyme variants were constructed and assayed for activity in Aspergillus niger as heterologous expression platform. We provide evidence that the β-ketoacyl synthase (KS) domain determines chain length in these mushroom NR-PKSs, even though their KS domains differ in only ten amino acids. A unique proline-rich linker connecting the acyl carrier protein with the thioesterase domain varies most between these two enzymes but is not involved in chain length control.     

Speciation of europium (III) surface species on monocrystalline alumina using time-resolved laser-induced fluorescence-scanning near-field optical microscopy

The aim of this work was to perform highly localized spectroscopic surface measurements by combining time-resolved laser spectroscopy and scanning near-field optical microscopy. The final purpose of that was to study surface sorption at the molecular level of trivalent ions in the framework of nuclear waste disposal assessment. Time-resolved laser spectroscopy presents the advantages of being selective, sensitive, and noninvasive and scanning near-field optical microscopy is a promising technique for high resolution surface speciation. Investigation of the interaction between trivalent europium and a monocrystalline alumina (1102) surface was made using different conditions of concentration and pH. We found that the distribution of sorbed europium was always homogeneous with a decay time of europium (III) equal to 350 micros+/-15 micros. On the other hand, carbonate species with a decay time of 210 micros+/-10 micros or other hydroxide species with a decay time of 180 micros+/-10 micros were detected on the surface when a higher concentration or a higher pH solution, respectively, were used. Distribution of these species was heterogeneous and their associated fluorescence signal was relatively high, evoking a precipitated form. X-ray photoelectron spectroscopy (XPS) was also used on the same samples as a complementary technique. A binding energy of 1135.1 eV was obtained for the sorbed europium and another binding energy of 1134.4 eV was obtained for the hydroxide species, thus confirming the presence of two kinds of species on the surface.     

In-situ electrical addressing of one-dimensional gold nanoparticle assemblies

Substrates with 1-dimensional nanosize grooves were prepared using extreme-ultraviolet interference lithography (EUV-IL), wherein gold nanoparticles were self-assembled to form 1-dimensional structures. To measure the electrical properties of gold nanoparticle chains we introduce a novel in-situ measuring method based on nanomanipulator system in a scanning electron microscope. This method comprises enormous versatility for the precisely electrical addressing of low-dimensional nanoscale structures and may even be applied to routinely addressing of structures in the sub-10 nm range.     

The fluorimetric FAST method, a simple tool for the optimization of microwave pasteurization of milk

The effect of microwave pasteurization of cow's milk on its nutritional quality was examined by the FAST method (Fluorescence of Advanced Maillard products and Soluble Tryptophan). Raw milk samples were submitted to different laboratory scale in batch microwave treatments using a central composite experimental design based on specific power and treatment time. The FAST index and bacterial count were monitored to assess protein denaturation, modification by the Maillard reaction and pasteurization efficiency, respectively. High discrimination between samples indicated that the FAST method is a potent tool for estimating the deterioration of the milk quality during experimental microwave treatment. Thus, the FAST index can be effectively used as the continuous response in experimental designs set up and to maximize information economically. In short, the FAST method allows us to retain the rapidity of experimental design while providing the advantages of convenience and low cost.     

Photonics based on carbon nanotubes

Among direct-bandgap semiconducting nanomaterials, single-walled carbon nanotubes (SWCNT) exhibit strong quasi-one-dimensional excitonic optical properties, which confer them a great potential for their integration in future photonics devices as an alternative solution to conventional inorganic semiconductors. In this paper, we will highlight SWCNT optical properties for passive as well as active applications in future optical networking. For passive applications, we directly compare the efficiency and power consumption of saturable absorbers (SAs) based on SWCNT with SA based on conventional multiple quantum wells. For active applications, exceptional photoluminescence properties of SWCNT, such as excellent light-emission stabilities with temperature and excitation power, hold these nanometer-scale materials as prime candidates for future active photonics devices with superior performances.     

Functional expression in insect cells, one-step purification and characterization of a recombinant phospholipase D from cowpea (Vigna unguiculata L. Walp)

Phospholipase D (PLD) is an important enzyme involved in signaltransduction, vesicle trafficking and membrane metabolism. Inthis study, large amounts of a recombinant plant PLD were secretedinto the culture medium of baculovirus-infected insect cellsand purified to homogeneity in the form of a fully active enzyme.The transient production of recombinant PLD yielded a protein(rPLDa, 88 kDa) together with a shorter form (rPLDb, 87 kDa),which accumulated in the medium. N-Terminal amino acid sequencingof the rPLDa and rPLDb showed that rPLDb resulted from proteolyticcleavage at Gly8–Ile9. Immunoblotting showed that bothrPLDa and rPLDb are recognized by a polyclonal antibody previouslyraised against native soybean PLD. One-step calcium-dependentoctyl-Sepharose chromatography was used to obtain the two highlypurified forms of rPLD, as attested by gel electrophoresis,N-terminal amino acid sequence and mass spectrometry. The N-terminalregion of PLD is homologous with the C2 domains which are presentin a number of enzymes known to be involved in signal transductionand/or phospholipid metabolism. The truncated rPLDb lacks thefirst acidic amino acid in its N-terminus, which is probablyinvolved in the calcium binding site. The rPLDb was thus easilyeluted from the octyl-Sepharose column by decreasing the calciumconcentration of the buffer from 50 to 30 mM, whereas, the rPLDawas eluted after chelating calcium ions with EDTA. The purifiedrPLD yield reached a level of 10 mg per liter of serum-freeculture medium. The availability of baculovirus-derived rPLDconstitutes a valuable source of enzyme for future crystallographicstudies to determine its three-dimensional structure.     

Causal attributions for school-related problems: Teacher perceptions and teacher feedback.

Conducted 2 studies with a total of 54 teachers to examine classroom teachers' attributions for severe school problems. In both studies, teachers who had referred a student for psychological services were asked to assign causality for the referral problem. In Exp II, teacher praise and criticism of referred students were examined as functions of causal attributions. Both studies show that teachers held student factors more responsible for classroom problems than teacher factors and that teachers' attributions varied somewhat for learning vs behavior problems. Exp II indicated that problem students perceived as lacking motivation were criticized more often by their teachers. Relationship of the results to attribution theory and teacher attribution research is discussed, and further research conducted in naturalistic settings is recommended. (22 ref) (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Strained Dehydrogenative Ring Closure of Phenylcarbazoles

What does it take to force a rigid and strained dehydrogenative ring closure, for example, in phenylcarbazoles? Since the works of Buchwald and Fagnou, palladium‐catalysed ring‐closing dehydrogenative reactions are legion, but will not operate when the strain at the reductive elimination stage becomes too large. We propose here a “muscled up” super‐oxidative palldium‐catalysed C H activation method for the ring closure of strained phenylcarbazoles.