Differentiation of Escherichia coli serotypes using DC gradient insulator dielectrophoresis

Bacteria play a significant role in both human health and disease. An estimated 9.4 million cases of foodborne illness occur in the United States each year. As a result, rapid identification and characterization of microorganisms remains an important research objective. Despite limitations, selective culturing retains a central role among a cadre of identification strategies. For the past decade, separations-based approaches to rapid bacterial identification have been under investigation. Gradient insulator dielectrophoresis (g-iDEP) promises benefits in the form of rapid and specific separation of very similar bacteria, including serotypes of a single species. Furthermore, this approach allows simultaneous concentration of analyte, facilitating detection and downstream analysis. Differentiation of three serotypes or strains of Escherichia coli bacteria is demonstrated within a single g-iDEP microchannel, based on their characteristic electrokinetic properties. Whole cells were captured and concentrated using a range of applied potentials, which generated average electric fields between 160 and 470 V/cm. Bacteria remained viable after exposure to these fields, as determined by cellular motility. These results indicate the potential g-iDEP holds in terms of both separatory power and the possibility for diagnostic applications.     

Chemical composition,antioxidant and cytotoxic activities of extracts from the leaves of Smilax brasiliensis Sprengel (Smilacaceae)

The antioxidant and cytotoxic activities of petroleum ether and methanol extracts, fatty acids and methyl esters from leaves of Smilax brasiliensis were evaluated, and the composition of the extracts was determined. Palmitic, linoleic and linolenic acids were major components of the extracts. For antioxidant activity, all samples exhibited IC₅₀ values lower than BHT (2,6-di-tert-butyl-4-methylphenol). The extracts, fatty acids and methyl esters from S. brasiliensis presented no toxicity to larvae of the brine shrimp, Artemia salina. Among the purified substances, only methyl linolenate showed toxicity (LD₅₀ = 21.47 μg/mL). This study showed, for the first time, the composition of petroleum ether and methanol extracts from S. brasiliensis leaves, as well as the antioxidant and cytotoxic activities of extracts, fatty acids and methyl esters.     

Synthesis and thermal crosslinking of carbosiloxane and oligo(oxyethylene) polymers

Acyclic diene metathesis (ADMET) polymerization has been used in the synthesis of telechelic materials using alkoxy‐functionalized carbosiloxane or oligo(oxyethylene)‐based polymers, varying from internal to terminal cured materials or the combination of them. Previous investigations demonstrated that introduction of chain‐end crosslinking improves the stress–strain behavior of such materials. A series of saturated and unsaturated carbosiloxane and oligo(oxyethylene)‐based polymers were synthesized by ADMET polymerization using silacyclobutane as chain‐end, thermally induced crosslinker. The carbosiloxane derivatives presented pure amorphous behavior, whereas the oligo(oxyethylene) polymers were semicrystalline. The thermal curing process was monitored by differential scanning calorimetry via the exotherm between 160 and 210 °C. Mechanical properties on thermoset polymers were measured, where cured polymers showed moduli from 0.6 to 9.3 MPa, tensile strengths from 0.3 to 1.0 MPa, and elongations from 12 to 76%. © 2011 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem, 2011     

High-Sensitivity Immunofluorescence Staining: A Comparison of the Liposome Procedure and the FASER Technique on mGR Detection

Flow cytometry has become a widely-used and powerful tool for the characterization of cells according to their expression of specific proteins. However, sensitivity of this method is still limited since conventionally labeled antibodies can be conjugated with at maximum 1–10 dye molecules. This fact resulted in the need to develop new techniques in order to identify molecules which are expressed in very low but functionally relevant amounts. In the past, we have successfully used a liposome-based high-sensitivity immunofluorescence technique to measure the expression of low abundant membrane bound glucocorticoid receptors (mGR) on different cell types. The use of this technique allows the detection of as few as 50–100 antigen molecules per cell which is due to a 100-fold to 1000-fold increase in fluorescence signal intensity compared with conventional methods. The higher sensitivity is achieved since thousands of dye molecules can be enclosed in liposomes. Another modern high-sensitivity immunofluorescence staining method is the purchasable Fluorescence Amplification by Sequential Employment of Reagents (FASER) procedure. Here, we aimed at comparing sensitivity and specificity of these two techniques for the detection of the mGR. Our data demonstrate the FASER technique to be more sensitive and also more specific for the detection of mGR as compared to the liposome technique. However, both methods have advantages and disadvantages which are discussed in detail.     

Inner-Sphere Oxygen Activation Promoting Outer-Sphere Nucleophilic Attack on Olefins

Alkoxylation and hydroxylation reactions of 1,5-cyclooctadiene (cod) in an iridium complex with alcohols and water promoted by the reduction of oxygen to hydrogen peroxide are described. The exo configuration of the OH/OR groups in the products agrees with nucleophilic attack at the external face of the olefin as the key step. The reactions also require the presence of a coordinating protic acid (such as picolinic acid (Hpic)) and involve the participation of a cationic diolefin iridium(III) complex, [Ir(cod)(pic)₂]⁺, which has been isolated. Independently, this cation is also involved in easy alkoxy group exchange reactions, which are very unusual for organic ethers. DFT studies on the mechanism of olefin alkoxylation mediated by oxygen show a low-energy proton-coupled electron-transfer step connecting a superoxide–iridium(II) complex with hydroperoxide–iridium(III) intermediates, rather than peroxide complexes. Accordingly, a more complex reaction, with up to four different products, occurred upon reacting the diolefin–peroxide iridium(III) complex with Hpic. Moreover, such hydroperoxide intermediates are the origin of the regio- and stereoselectivity of the hydroxylation/alkoxylation reactions. If this protocol is applied to the diolefin–rhodium(I) complex [Rh(pic)(cod)], free alkyl ethers ORC₈H₁₁ (R=Me, Et) resulted, and the reaction is enantioselective if a chiral amino acid, such as l -proline, is used instead of Hpic.     

Enantiodivergence in the reduction of α-methyl and α-halomethyl enones by microorganisms

Enones (Z)-3-methyl-(Z)-3-chloromethyl- and (Z)-3-bromomethyl-4-R-3-buten-2-one (R = n-pentyl, phenyl, 2′- and 4′-chlorophenyl, 3′- and 4′-nitrophenyl, 4′-methoxyphenyl) were synthesized and subjected to reduction by the microorganisms Saccharomyces cerevisiae andGeotrichum candidum. Whereas the bioreduction of 3-methy-4-R-3-buten-2-ones afforded the corresponding (S)-4-R-3-methybutan-2-ones, the bioreduction of 3-chloromethyl- and 3-bromomethyl-4-R-3-buten-2-ones afforded the corresponding (R)-4-R-3-methybutan-2-ones.     

Highly Selective Zeolite Topologies for Flue Gas Separation

The separation of carbon dioxide from flue gas is essential for the reduction of greenhouse gas emissions. In adsorptive methods, the challenge lies in the choice of suitable porous materials. Among all zeolite topologies, a number of adsorbents with pore dimensions in the range of the guest molecules were identified to allow an excellent separation by diffusion, and MRE and AFO zeolite topologies appear to be the best candidates based on equilibrium adsorption. Also, it was found that the behavior of this gas mixture in DFT and APD zeolites differed from the normal behavior.     

Amorphous Calcium Phosphates: Solvent-Controlled Growth and Stabilization through the Epoxide Route

Calcium phosphates stand among the most promising nanobiomaterials in key biomedical applications, such as bone repairment, signalling or drug/gene delivery. Their intrinsic properties as crystalline structure, composition, particle shape and size define their successful use. Among these compounds, metastable amorphous calcium phosphate (ACP) is currently gaining particular attention due to its inherently high reactivity in solution, which is crucial in bone development mechanisms. However, the preparation of this highly desired (bio)material with control over its shape, size and phase purity remains as a synthetic challenge. In this work, the epoxide route was adapted for the synthesis of pure and stable ACP colloids. By using biocompatible solvents, such as ethylene glycol and/or glycerine, it was possible to avoid the natural tendency of ACP to maturate into more stable and crystalline apatites. Moreover, this procedure offers size control, ranging from small nanoparticles (60 nm) to micrometric spheroids (>500 nm). The eventual fractalization of the internal mesostructured can be tuned, by simply adjusting the composition of the ethylene glycol:glycerine solvent mixture. These findings introduce the use of green solvents as a new tool to control crystallinity and/or particle size in the synthesis of nanomaterials, avoiding the use of capping agents and preserving the natural chemical reactivity of the pristine surface.