Cancer‐Cell‐Specific Induction of Apoptosis Using Mesoporous Silica Nanoparticles as Drug‐Delivery Vectors

Targeted delivery of the chemotherapeutic agent methotrexate (MTX) to cancer cells using poly(ethyleneimine)‐functionalized mesoporous silica particles as drug‐delivery vectors is reported. Due to its high affinity for folate receptors, the expression of which is elevated in cancer cells, MTX serves as both a targeting ligand and a cytotoxic agent. Enhanced cancer‐cell apoptosis (programmed cell death) relative to free MTX is thus observed at particle concentrations where nonspecific MTX‐induced apoptosis is not observed in the nontargeted healthy cell line, while corresponding amounts of free drug affect both cell lines equally. The particles remain compartmentalized in endo‐/lysosomes during the time of observation (up to 72 h), while the drug is released from the particle only upon cell entry, thereby inducing selective apoptosis in the target cells. As MTX is mainly attached to the particle surface, an additional advantage is that the presented carrier design allows for adsorption (loading) of additional drugs into the pore network for therapies based on a combination of drugs.     

High-throughput screening of protein surface activity via flow injection analysis-pH gradient-dynamic surface tension detection

Using flow injection analysis (FIA), a pH gradient is blended in real time with a protein sample as the pH-dependent protein surface activity is measured by a dynamic surface tension detector (FIA-pH-DSTD). This instrumental system was developed as a high-throughput method for the screening of protein surface activity at the air/liquid interface as a function of pH. This method utilizes the continuous flow, drop-based dynamic surface tension detector in combination with flow injection sample introduction and blending of a steady-state concentration of protein sample with a pH gradient ranging from pH 2.0 to pH 11.5. Dynamic surface tension is measured through the differential pressure across the air/liquid interface of repeatedly growing and detaching drops. Continuous surface tension measurement is achieved for each eluting drop of 2-s length (2 muL), providing insight into both the kinetic and thermodynamic behaviors of molecular orientation processes at the liquid/air interface. Three-dimensional data are obtained, with surface tension first converted to surface pressure, which is collected as a function of elution time versus drop time. In FIA-pH-DSTD, a commercial pH probe is used to measure pH during elution time, enabling surface pressure throughout drop time to be subsequently plotted as a function of eluting pH. An automated DSTD calibration procedure and data analysis method is applied, which allows simultaneous use of two different solvents, permitting real-time dynamic surface tension data to be obtained. The method was applied to the analysis of 14 commercial purified proteins, yielding characteristic features of surface activity as a function of pH. The reproducibility of the measurement and selectivity advantage of the DSTD was shown for the analysis of serum albumins from various mammalian sources. Several applications were also suggested and discussed in order to show the potential of the method for protein and food chemistry studies and in the study of protein-polymer interactions.     

Microwave-assisted photochemical reactor for the online oxidative decomposition and determination of p-hydroxymercurybenzoate and its thiolic complexes by cold vapor generation atomic fluorescence detection

We developed a photochemical method for the online oxidation of p-hydroxymercurybenzoate (PHMB), an organic mercury species widely used for mercaptan and thiolic compound labeling. The method is based on a fully integrated online UV/microwave (MW) photochemical reactor for the digestion of PHMB, followed by cold vapor generation atomic fluorescence spectrometry (CVG-AFS) detection. The MW/UV process led to the quantitative conversion of PHMB and thiol-PHMB complexes to Hg(II), with a yield between 91% and 98%, without using chemical oxidizing reagents and avoiding the use of toxic carcinogenic compounds. This reaction was followed by the reduction of Hg(II) to Hg(0), performed in a knitted reaction coil with NaBH(4) solution, and AFS detection in an Ar/H(2) miniaturized flame. The low MW power applied (18 W) allowed us to keep constant the temperature of the photochemical reactor (21 ± 1 °C), using a flowing water bath. This avoided peak widening due to diffusion processes generally occurring at high temperatures and in the additional cooling coil. This method has been applied to the determination of thiols in human plasma, blood, and wine.     

Interaction of erythrocytes with magnetic nanoparticles

Internalization of biocompatible magnetic nanoparticles by red blood cells (RBCs) is a key issue for opportunities of new applications in the biomedical field. In this study, we used in vitro tests to provide evidences of magnetic nanoparticle internalization by mice red blood cells. The internalization process depends upon the nanoparticle concentration and the nanoparticle hydrodynamic radii. The cell internalization of surface-coated maghemite nanoparticles was indirectly tracked by Raman spectroscopy and directly observed using transmission electron microscopy. The observation of nanoparticle cell uptaking using in vitro experiments represents an important breakthrough for the application of nanomagnetism in diagnosis and therapy of RBC-related diseases.     

Structural and spectroscopic characteristics of synthetic hydrohaematite

In order to investigate the identity of water-containing haematite, a method of preparation has been elaborated which leads to samples of hydrohaematite which are structurally pure and free of amorphous iron hydroxide. Differential thermal analysis by a lack of endothermic effect at 423 to 473 K on the DTA-curves and by a steady fall of the TG curves up to 1150 K revealed that water in the preparations must be tightly held in the lattice of haematite. Measurements of intensities of X-ray reflections of the (1 0 4) and (0 2 4) planes in relation to the (1 1 3) plane confirmed Fe³⁺-deficiency in the haematite cationic sublattice brought about by the presence of OH^– ions in the anionic sublattice. The infrared spectrum of hydrohaematite, in addition to six bands from Fe-O variations, exhibits three bands from hydroxyl groups. The effect of silicates on the pattern of the infrared spectra of natural and synthetic hydrohaematites and the discrepancies in the infrared spectra of haematites published to date are discussed.     

Development and validation of a high-performance liquid chromatography-mass spectrometry assay for determination of amphetamine, methamphetamine, and methylenedioxy derivatives in meconium

A procedure based on liquid chromatography-mass spectrometry (LC-MS) is described for determination of amphetamine, methamphetamine, and methylendioxy derivatives in meconium, using 3,4-methylendioxypropylamphetamine as internal standard. The analytes were initially extracted from the matrix by 17 mM methanolic HCl. Subsequently, a solid-phase extraction with Bondelut Certify columns was applied. Chromatography was performed on a C(18) reversed-phase column using a linear gradient of 10 mM ammonium bicarbonate, pH 9.0-methanol as a mobile phase. Analytes were determined in LC-MS single ion monitoring mode with an atmospheric pressure ionization-electrospray interface. The method was validated in the range 0.005-1.00 microg/g using 1 g of meconium per assay. Mean recoveries ranged between 61.1 and 87.2% for different analytes. The quantification limits were 0.005 microg/g meconium for amphetamine, methamphetamine, and 4-hydroxy-3-methoxymethamphetamine and 0.004 microg/g meconium for 3,4-methylenedioxyamphetamine, 3,4-methylenedioxymethamphetamine, 3,4-methylenedioxyethylamphetamine, and N-methyl-1-(3,4-methylenedioxyphenyl)-2-butanamine. The method was applied to analysis of meconium in newborns to assess eventual fetal exposure to amphetamine derivatives.     

The influence of powder morphology on the effect of rosemary aroma microencapsulation during spray drying

The influence of particle morphology of spray dried powders obtained by using different carriers on the efficiency of microencapsulation of rosemary aroma is shown in the present research. It was found that the type of the carrier influenced the size but not the shape of rosemary aroma capsules and all the capsules were spherical with cracks. The biggest particles, d₅₀ = 55 μm, were obtained with 25% maltodextrin (MD) as a carrier, and the smallest, d₅₀ = 29 μm with 30% gum arabic (GA). The efficiency of encapsulating aroma inside of the capsules depended on the particles size and apparent density. An increase in the quantity of microencapsulated aroma from 10% for 25% MD to about 60% for 30% GA as the carrier was seen, which co‐related with a decrease in the average diameter of the powder particles. Similarly, an increase in the efficiency of aroma retention was followed by an increase of apparent density of powders from 796 (25% MD) to 1156 kg m^?3 (30% GA).     

Determination of 4-hydroxynonenal by high-performance liquid chromatography with electrochemical detection

4-Hydroxy-trans-2-nonenal (HNE) is a highly reactive product of lipid peroxidation originating from the breakdown of phospholipid-bound polyunsaturated fatty acids of cellular membranes. Despite its biological relevance, this aldehyde is only occasionally determined due to the complexity of previously described procedures. Here we present a simple and very sensitive method for the detection of HNE in biological samples. The method is based on the measurement of the 2,4-dinitrophenylhydrazone (DNPH) of the aldehyde by electrochemical detection after separation by reverse-phase high-performance liquid chromatography (HPLC). The greater sensitivity of this procedure as compared to the ultraviolet detection method commonly employed to measure DNPH derivatives of aldehydes after HPLC will allow the detection of HNE below the pmol level. The detection of HNE is highly reproducible even in normal tissues and cells. Increased amounts of HNE were detected in the livers of animals intoxicated with prooxidant agents such as carbon tetrachloride, bromotrichloromethane or bromobenzene. An exponential increase in HNE (and in malondialdehyde) was measured in peroxidizing liver microsomes (in the NADPH/Fe-dependent system). The method is also suitable for the study of very small samples, since HNE could be detected in approximately 1 million cultured cells (polyoma virus-transformed baby hamster kidney fibroblasts); the level rose after exposure of the cells to a Fe³⁺/ADP prooxidant system. During the course of these studies, Dr. Goldring was on leave from Division of Biochemistry, UMDS Guy's Hospital, London SE1 9RT, UK.     

Efficient UV polymerisation of 3BCMU: Optical and waveguiding properties of the material

A modified method to synthesize 3BCMU and to polymerize it by ultraviolet (UV) radiation is presented; the polymerization technique gave a maximum yield of 18%. Langmuir-Blodgett and spin coated films of UV/poly-3BCMU have been fabricated. Visible and resonance Raman spectroscopy and waveguide tests were performed: the results are compared with those obtained with y/poly-3BCMU films.     

Continuous biotransformation of pyrogallol to purpurogallin using cross‐linked enzyme crystals of laccase as catalyst in a packed‐bed reactor

Cross‐linked enzyme crystals (CLEC) of laccase were prepared by crystallizing laccase with 75% (NH₄)₂SO₄ and cross‐linking using 1.5% glutaraldehyde. The cross‐linked enzyme crystals were further coated with 1 mmol L^?1 β‐cyclodextrin by lyophilization. The lyophilized enzyme crystals were used as such for the biotransformation of pyrogallol to purpurogallin in a packed‐bed reactor. The maximum conversion (76.28%) was obtained with 3 mmol L^?1 pyrogallol at a residence time of 7.1 s. The maximum productivity (269.03 g L^?1 h^?1) of purpurogallin was obtained with 5 mmol L^?1 pyrogallol at a residence time of 3.5 s. The productivity was found to be 261.14 g L^?1 h^?1 and 251.1 g L^?1 h^?1 when concentrations of 3 mmol L^?1 and 7 mmol L^?1 respectively were used. The reaction rate of purpurogallin synthesis was maximum (2241.94 mg purpurogallin mg^?1 CLEC h^?1) at a residence time of 3.5 s, when 5 mmol L^?1 pyrogallol was used as the substrate. The catalyst to product ratio calculated for the present biotransformation was 1:2241. The CLEC laccase had very high stability in reuse and even after 650 h of continuous use, the enzyme did not lose its activity. Copyright © 2006 Society of Chemical Industry