USING CELL-FRACTIONATION AND PHOTOCHEMICAL CROSSLINKING METHODS TO DETERMINE THE CELLULAR BINDING SITE(S) OF THE ANTITUMOR DRUG DMP 840

Abstract— In order to understand its mechanism of action we have begun an effort to better define the cellular target of action of the experimental antitumor agent DMP 840 (NSC D640430; (R, R)-2,2'-(1,2-ethanediylbis(imino-(1 -methyl-2,1 -ethanediyl)))-bis(5-nitro-1 H-benz(de)isoquinoline-1,3-(2H)-dione) dimethanesulfonate). Using a combination of gentle cell fractionation procedures and a previously unidentified photochemical crosslinking reaction, we have shown that after the drug is added to cultured Clone A cells, more than 80% of the drug that is found associated with cells partitions to the chromatin-containing structural framework of the cell and that the primary target after crosslinking with 360 nm light is DNA. While DMP 840 photoreacts quite efficiently with purified RNA in vitro , no photoat-tachment of the drug to RNA was observed in cells. In vitro photochemical studies also reveal that while GC-rich DNA is a preferred target for drug interaction, AT-rich DNA is more active in the photochemical crosslinking reaction. These results suggest that DMP 840 probably kills cells by interfering with DNA-metabolic processes, and that the drug and its derivatives are likely to be useful photoactive molecular probes for investigating higher order chromatin structures in cells.     

Necessary Optimality Conditions for Higher-Order Infinite Horizon Variational Problems on Time Scales

We obtain Euler?CLagrange and transversality optimality conditions for higher-order infinite horizon variational problems on a time scale. The new necessary optimality conditions improve the classical results both in the continuous and discrete settings: Our results seem new and interesting even in the particular cases when the time scale is the set of real numbers or the set of integers.     

Evaluation of graphene oxide and reduced graphene oxide in the immobilization of laccase enzyme and its application in the determination of dopamine

Bioelectrodes were developed based on a simple deposition of graphene oxide (GO) or reduced graphed oxide (rGO) and laccase (Lac) on a glassy carbon (GC) electrode surface. The morphology and electrochemical behavior of the biosensors were characterized by scanning electron microscopy and cyclic voltammetry. These results demonstrated that only rGO was successfully applied for the immobilization of the laccase enzyme, improving the analytical signal for the determination of dopamine. The GC/rGO/Lac biosensor was applied to the detection of dopamine in synthetic urine and plasmatic serum samples, achieving a detection limit of 91.0 nmol L^?1.     

Voltammetric insights in the transfer of ionizable drugs across biomimetic membranes: recent achievements

The latest results of voltammetric research on the ionic transfer process of ionisable drugs across bare and lipid-modified liquid-liquid interfaces are reviewed. In recent years, two voltammetric methods have been extensively applied to this purpose, i.e. the classical four electrode voltammetry at the interface between two immiscible electrolyte solutions, and the "three-phase electrode." Thus, a brief background of the methodologies and some successful examples of their application are highlighted in this work. Particular attention is given to the ionic transfer kinetics and to the electrochemical characterization of the drug-membrane interactions between the ionized drugs and lipid-modified interfaces. Future trends in this area are also mentioned in connection with high-throughput assessment of ADMET properties of drugs.     

Rapid determination of gold in Romanian auriferous alluvial sands, concentrates and rocks by 14 MeV NAA

The nuclear and spectral interferences in the 14 MeV neutron activation analysis (NAA) of gold from Romanian auriferous alluvial sands, concentrates and rocks have been studied and the optimum of activation, cooling and measuring times was determined for a maximum peak-to-background ratio for gold. The contribution of the nuclear interfering elements in the samples, Hg and Pt, to the concentration of gold has been calculated and, concluded that the nuclear reactions ¹⁹⁷Au(n,2n)¹⁹⁶Au, ¹⁹⁷Au(n,2n)^196mAu and ¹⁹⁷Au(n,n')^197mAu can be used for gold determination, with minimal errors. Using the nuclear reactions ¹⁹⁷Au(n,n')^197mAu and ¹⁹⁷Au(n,2n)¹⁹⁶Au the spectral interferences are minimal and are due to Rb, Ti and V for a short irradiation and to Se for a long one. Two methods of fast gold determination were proposed for auriferous alluvial sands and rocks in the range of 20-2500 ppm, under the optimum conditions established so that the systematic errors of analysis due to the gold accompanying elements can be considerably diminished. For measuring the induced gamma-radioactivity in the samples either a short irradiation (25 seconds) with a NaI(Tl) detector or a long irradiation (3000 seconds) with a Ge(Li) detector were used. This revised version was published online in July 2006 with corrections to the Cover Date.     

Hemisynthesis and structural characterization of flavanol‐(4,8)‐vitisins by mass spectrometry

Vitisins constitute the major group of pyranoanthocyanins naturally occurring in red wines. Regarding their chemical structure, only carboxypyranoanthocyanins have been detected and quantified in red wines, but no vitisin with substitutions in the carbons of the A ring has been identified. However, considering the chemical reactions that take place in red wine, the existence of flavan‐3‐ol‐(4,6/8)‐vitisins is expected. This paper reports for the first time the structural identification of catechin‐vitisin A and catechin‐vitisin B based on their mass data and fragmentation patterns. This work also provides some chromatographic and visible spectroscopic features of these pigments and documents the existence of both in red table wines. Moreover, it is also proposed that Cat‐Vit A pigments arise from the cycloaddition of pyruvic acid to an anthocyanin moiety of a flavanol‐anthocyanin adduct rather than by direct nucleophilic attack of a vitisin A on the carbocation C₄ of catechin. Copyright © 2010 John Wiley & Sons, Ltd.     

Cytotoxic compounds from the marine-derived fungus Aspergillus sp. recovered from the sediments of the Brazilian coast

A fungal strain of Aspergillus sp. (BRF 030) was isolated from the sediments collected in the northeast coast of Brazil, and the cytotoxic activity of its secondary metabolites was investigated against HCT-116 tumour cell line. The cytotoxicity-guided fractionation of the extracts from this fungus cultured in potato-dextrose-sea water for 14 days at room temperature yielded the hetero-spirocyclic γ-lactams pseurotin A (1), pseurotin D (2) and pseurotin FD-838 (7), the alkaloids fumitremorgin C (5), 12,13-dihydroxy fumitremorgin C (6), methylsulochrin (4) and bis(dethio)bis(methylthio)gliotoxin (3). Among them, fumitremorgin C (5) and 12,13-dihydroxy fumitremorgin C (6) were the most active. The cytotoxic activities of the extracts from Aspergillus sp. grown from 7 to 28 days were investigated, and they were associated with the kinetic production of the compounds. The most active extracts (14 and 21 days) were those with the highest relative concentrations of the compounds fumitremorgin C (5) and 12,13-dihydroxy fumitremorgin C (6).     

N,N′-bridged binuclear NHC palladium complexes: A combined experimental catalytic and computational study for the Suzuki reaction

This work examines how N-donor bridged spacer ligands affect N-heterocyclic carbene (NHC) palladium complexes catalytic activities for Suzuki coupling reaction. Different degrees of structural flexibility binuclear NHC palladium complexes were synthesized. The more flexible nitrogen-based alkyl chain ligand shows similar performance with cycloamine counterparts in the Suzuki coupling reaction. Suzuki coupling examples were used in air and ambient temperature to reach moderate to completion yields in short time. Density functional theory calculations showed that the chelate effect, associated with a single Pd complex mechanism, plays a fundamental role in the pre-catalysis stage, supporting a reasonable of the kinetic activity observed experimentally.     

What does an ionic liquid surface really look like? Unprecedented details from molecular simulations

We present the first intrinsic analysis of the surface of the [bmim][PF(6)] room-temperature ionic liquid. Our detailed analysis reveals unprecedented details about the structure of the interface by providing the relative prevalence of different molecular orientations. These results suggest that experimental data should be reinterpreted considering a distribution of molecular arrangements.