The application of diethylaminosulfur trifluoride in the synthesis of fluorinated sterols and bile acids

Methyl 3,3-(ethylenedioxy)-6β-fluoro-5β-cholan-24-oate has been prepared starting from hyodeoxycholic acid and 3,3-(ethylenedioxy)-7( and β)-fluorocholestan-22-al starting from stigmasterol, using diethylaminosulfur trifluoride as the fluorinating agent. These molecules will be used to synthesize analogs of the aminosterol antibiotic squalamine.     

Tuning cell adhesion on gradient poly(2-hydroxyethyl methacrylate)-grafted surfaces

A simple yet versatile method was developed to prepare a low-density polymerization initiator gradient, which was combined with surface-initiated atom transfer radical polymerization (ATRP) to produce a well-defined poly(2-hydroxyethyl methacrylate) (HEMA) gradient substrate. A smooth variation in film thickness was measured across the gradient, ranging from 20 A to over 80 A, but we observed a nonmonotonic variation in water contact angle. Fits of X-ray reflectivity profiles suggested that at the low graft density end, the polymer chain structure was in a "mushroom" regime, while the polymer chains at high graft density were in a "brush" regime. It was found that the "mushroom" region of the gradient could be made adhesive to cells by adsorbing adhesion proteins, and cell adhesion could be tuned by controlling the density of the polymer grafts. Fibroblasts were seeded on gradients precoated with fibronectin to test cellular responses to this novel substrate, but it was found that cell adhesion did not follow the expected trend; instead, saturated cell adhesion and spreading was found at the low grafting density region.     

Synthesis, characterization, and preliminary intramolecular energy transfer studies of rigid, emissive, rhenium-linked porphyrin dimers

The reaction of pyridyl functionalized porphyrins with Re(CO)(5)Cl in THF results in the formation of porphyrin dimers which, despite incorporation of rhenium into the assemblies, remain fluorescent. The rigid compounds provide an efficient geometry and/or orbital pathway for singlet energy transfer, rendering these compounds suitable, in principle, for the study of both through-bond and through-space energy transfer. Derivatives containing both metallated and freebase porphyrins connected via the metal corners display efficient porphyrin-porphyrin energy transfer. The photophysical properties of the assemblies have been studied by both steady-state and time-resolved fluorescence techniques, yielding approximate rates and efficiencies for porphyrin-porphyrin energy transfer.     

Neutron activation analysis of zeolite catalysts

A new method for the determination of aluminum and silicon has been developed for zeolite catalysts. In contrast to previous methods, thermal neutrons are used for the analysis of both elements, and cadmium absorbers are not needed. The silicon determination utilizes a one-hour irradiation to observe the³¹Si produced by the (n, ) reaction of³⁰Si. A 15-second irradiation is used for the²⁷Al(n, )²⁸Al reaction. The²⁸Al activity is corrected for the contribution from the²⁸Si(n,p)²⁸Al reaction by using the analyzed weight of silicon in the sample and the data for a silicon standard irradiated simultaneously with the zeolite and the aluminum standard. The quantitation limits are 0.012 g for silicon and 3.3×10^–5 g for aluminum. Sodium presents a significant interference, but this element can be removed by taking advantage of the ion exchange properties of these materials.     

Oxygen plasma-treatment effects on Si transfer

Oxygen plasma-treatment is commonly used to increase the hydrophilicity of poly(dimethylsiloxane) (PDMS) stamps used for microcontact printing (muCP) aqueous-based inks. Review of the literature reveals that a wide range of plasma parameters are currently employed to modify stamp surfaces. However, little is known about the effect of these parameters (e.g., power, chamber pressure, duration) on the undesirable transfer of low-molecular-weight silicon-containing fragments from the stamps that commonly occurs during muCP. To study the effect of oxygen plasma-treatment on Si transfer, unpatterned PDMS stamps were treated with oxygen plasma under various conditions and used to stamp deionized water on plasma-activated poly(methyl methacrylate) (PMMA) substrates. Once stamped, the PMMA substrates were analyzed with X-ray photoelectron spectroscopy (XPS) to quantify and characterize silicon present on the substrate surface. In addition, used PDMS stamps were analyzed with scanning electron microscopy (SEM) to observe topographical changes that occur during oxygen plasma-treatment. XPS results show that all plasma treatments studied significantly reduced the amount of Si transfer from the treated stamps during muCP as compared to untreated PDMS stamps and that the source of transfer is residual PDMS fragments not removed by oxygen plasma. SEM results show that, although the treated stamps undergo a variety of topographical changes, no correlation exists between stamp topography and extent of Si transfer from the stamps.     

SITES OF PHOTODAMAGE in vivo and in vitro BY A CATIONIC PORPHYRIN

Abstract— Localization and photodynamic efficacy of a monocationic porphyrin (MCP) were assessed using murine leukemia cells in culture. This sensitizer localized at surface membrane loci and catalyzed selective photodamage to membrane structures. Although both cationic and hydrophobic, this porphyrin was not recognized by the multidrug transporter, which excludes many cationic agents from cells that express multidrug resistance. Photodynamic studies with the murine radiation-induced fibrosarcoma tumor model indicated moderate photosensitization of neoplastic lesions in vivo at 3 h, but not at 24 h after sensitizer administration. Pharmacokinetic studies indicate that plasma levels, not tissue levels were the major determinant of photodynamic therapy (PDT) response. Consistent with this observation, vascular damage and disturbances of tissue perfusion followed PDT. These effects were more pronounced in tumor-bearing skin than in normal skin. The therapeutic response to MCP appeared to be related mainly to secondary, probably vascular, effects.     

Estrogen receptor microarrays: subtype-selective ligand binding

We present the first example of a nuclear hormone receptor microarray, using for illustration the ligand-binding domains of the two estrogen receptors, ERalpha-LBD and ERbeta-LBD. The proteins are printed and allowed to attach to aldehyde slides; the efficiency of attachment depends on whether the LBD is liganded with agonists (low attachment) versus liganded with antagonists or unliganded (high attachment). This suggests that attachment is orientation specific and involves principally a single lysine residue. The attached ERs retain good ligand-binding activity that can be assessed using an estradiol-fluorophore conjugate, and specific and ER subtype-selective binding of ligands can be determined conveniently in competitive binding assays. This powerful new, high-throughput technique to study ligand binding to ER-LBDs can be extended to other nuclear hormone receptors and adapted to assay the recruitment of coregulator proteins.     

Selenium redox cycling in the protective effects of organoselenides against oxidant-induced DNA damage

The biological role of selenium is a subject of intense current interest, and the antioxidant activity of selenoenzymes is now known to be dependent upon redox cycling of selenium within their active sites. Exogenously supplied or metabolically generated organoselenium compounds, capable of propagating a selenium redox cycle, might therefore supplement natural cellular defenses against the oxidizing agents generated during metabolism. We now report evidence that selenium redox cycling can enhance the protective effects of organoselenium compounds against oxidant-induced DNA damage. Phenylaminoethyl selenides were found to protect plasmid DNA from peroxynitrite-mediated damage by scavenging this powerful cellular oxidant and forming phenylaminoethyl selenoxides as the sole selenium-containing products. The redox properties of these organoselenoxide compounds were investigated, and the first redox potentials of selenoxides in the literature are reported here. Rate constants were determined for the reactions of the selenoxides with cellular reductants such as glutathione (GSH). These kinetic data were then used in a MatLab simulation, which showed the feasibility of selenium redox cycling by GSH in the presence of the cellular oxidant, peroxynitrite. Experiments were then carried out in which peroxynitrite-mediated plasmid DNA nick formation in the presence or absence of organoselenium compounds and GSH was monitored. The results demonstrate that GSH-mediated redox cycling of selenium enhances the protective effects of phenylaminoethyl selenides against peroxynitrite-induced DNA damage.     

Dodecyl sulfate-hydrotalcite nanocomposites for trapping chlorinated organic pollutants in water

A series of hybrid organic-inorganic nanocomposite materials was synthesized by three different procedures using sodium dodecyl sulfate (DDS) and magnesium-aluminum layered double hydroxide (Mg/Al LDH with a Mg/Al molar ratio of 2 to 5). Both the pH of the exchange medium (6.5 to 10) and the Mg/Al molar ratio of the LDH affected the basal spacing, the content of DDS retained and the orientation of the DDS chains within the interlamellar space. For LDH with higher charge density (Mg/Al=2 and 3), DDS molecules likely formed a perpendicular monolayer within the LDH interlayer and the solution pH had little effect on the basal spacing, with a mean and standard deviation of 25.5+/-0.4 A. However, for LDH with lower charge density (Mg/Al=4 and 5), DDS molecules more likely formed an interpenetrating bilayer, and the basal spacing significantly increased with increasing pH, with a mean and standard deviation of 32.7+/-5.2 A. Sorption of trichloroethylene and tetrachloroethylene by DDS-LDH varied with synthesis conditions, LDH type and DDS configuration in the interlayer. DDS-Mg(3)Al-LDH had the highest affinity for both trichloroethylene and tetrachloroethylene in water, either comparable to or as much as four times higher than other clay-derived sorbents, followed by DDS-Mg(4)Al-LDH and DDS-Mg(5)Al-LDH. DDS-Mg(2)Al-LDH had the lowest sorption affinity although the highest amount of DDS. The pH of the exchange solution also affected the amount of DDS retained by the LDH as well as the sorption efficiency. Mg(3)Al-LDH has a charge equivalent area of 32.2 A(2)/charge, which allows the formation of optimal DDS configuration within its interlayer, thus resulting in the highest affinity for the chlorinated compounds. The DDS-Mg/Al-LDHs can be easily synthesized either ex situ or in situ at low temperature, indicating the feasibility of practical applications. The results obtained by controlling the synthesis procedure suggest that different arrangements of DDS molecules in the LDH interlayers can be obtained and optimized for the sorption of specific sorbates.     

Fragmentation of N-linked glycans with a matrix-assisted laser desorption/ionization ion trap time-of-flight mass spectrometer

N-Linked glycans were ionized from several matrices with a Shimadzu-Biotech AXIMA-QIT matrix-assisted laser desorption/ionization quadrupole ion trap time-of-flight mass spectrometer. [M+Na]+ ions were produced from all matrices and were accompanied by varying amounts of in-source fragmentation products. The least fragmentation was produced by 2,5-dihydroxybenzoic acid and the most by alpha-cyano-4-hydroxycinnamic acid and 6-aza-2-thiothymine. Sialic acid loss was extensive but could be prevented by formation of methyl esters. Fragmentation produced typical low-energy-type spectra dominated by ions formed by glycosidic cleavages. MS(n) spectra (n = 3 and 4) were used to probe the pathways leading to the major diagnostic ions. Thus, for example, an ion that was formed by loss of the core GlcNAc residues and the 3-antenna was confirmed as being formed by a B/Y rather than a C/Z mechanism. The proposed structures of several cross-ring cleavage ions were confirmed and it was shown that MS3 spectra could be obtained from as little as 10 fmol of glycan.