Photoactivation of pheophorbide a induces a mitochondrial-mediated apoptosis in Jurkat leukaemia cells.

The mechanism of cell death by pheophorbide a (Pba) which has been established to be a potential photosensitizer was examined in experimental photodynamic therapy (PDT) on Jurkat cells, a human lymphoid tumor cell line. In 30-60 min after irradiation, Pba treated cells exhibited apoptotic features including membrane blebbing and DNA fragmentation. Pba/PDT caused a rapid release of cytochrome c from mitochondria into the cytosol. Sequentially, activation of caspase-3 and the cleavage of poly ADP-ribose polymerase (PARP) were followed. Meanwhile, no evidence of activation of caspase-8 was indicated in the cells. In experiments with caspase inhibitors, it was found that caspase-3 alone was sufficient initiator for the Pba-induced apoptosis of the cells. Pba specific emission spectra were confirmed in the mitochondrial fraction and the light irradiation caused a rapid change in its membrane potential. Thus, mitochondria were entailed as the crucial targets for Pba as well as a responsible component for the cytochrome c release to initiate apoptotic pathways. Taken together, it was concluded that the mode of Jurkat cell death by Pba/PDT is an apoptosis, which is initiated by mitochondrial cytochrome c release and caspase-3-pathways.

Polychlorinated dibenzo-p-dioxins and dibenzofurans from food salt samples processed by heat treatment in Korea.

Eight samples of processed food salt collected from five plants in Korea were analyzed for 2,3,7,8-substituted polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) using liquid-liquid extraction, clean-up procedures, and high resolution gas chromatography-high resolution mass spectrometry. The study included the analyses of two kinds of salt product sample: bamboo-salt and parched salt. The levels of toxic PCDD/Fs found in the salt product samples were extremely low: the results revealed TEQ levels ranging between the sub pg TEQ/g and sub fg TEQ/g. The differences in the TEQ values of toxic PCDD/F were observed between the salt product samples, which were treated with different frequency of baking using four different fuels (firewood, pine wood, pine resin, and indirect heating by gas) at temperatures from 300 to 2000 degrees C. In bamboo-salt samples, the concentrations and TEQ values of toxic PCDD/Fs ranged between 0.57-66 pg/g and 5.7x10(-5)-0.64 pg TEQ/g, respectively. PCDD/Fs levels in bamboo-salts baked by firewood were found to be higher than those baked by pinewood or pine resin. In parched salt samples, the concentrations and TEQ values of toxic PCDD/Fs ranged between 0.97-3.7 pg/g and 0.0097-0.017 pg TEQ/g, respectively. The data was discussed regarding the concentration and the distribution pattern of congeners.

Size-based analysis of incinerator fly ash using gravitational SPLITT fractionation, sedimentation field-flow fractionation, and inductively coupled plasma-atomic emission spectroscopy.

Fly ash has been regarded as hazardous because of its high adsorption of toxic organic and/or inorganic pollutants. Fly ash is also known to have broad distributions of different chemical and physical properties, such as size and density. In this study, fly ash emitted from a solid waste incinerator was pre-fractionated into six sub-populations by use of gravitational SPLITT fractionation (GSF). The GSF fractions were then analyzed by sedimentation field-flow fractionation (SdFFF) and ICP-AES. SdFFF analysis showed the fly ash has a broad size distribution ranging from a few nanometers up to about 50 microm. SdFFF results were confirmed by electron microscopy. Inductively coupled plasma-atomic emission spectroscopy (ICP-AES) analysis of the GSF fractions showed the fly-ash particles contain a variety of inorganic elements including Ca, Si, Mg, Fe, and Pb. The most abundant in fly ash was Ca, followed by Si then Mg. No correlations were found between trace element concentration and particle size.

Size analysis of industrial carbon blacks by sedimentation and flow field-flow fractionation.

Carbon black is one of the most useful particulate materials in the industrial field. Among the various physical properties of carbon black, size and size distribution are the most important properties to affect the quality of a final product. However, it is difficult to measure the exact particle size of carbon black since it suffers unavoidable interference from flocculation. In this study, the effects of various factors on the dispersion of industrial carbon blacks were investigated for the determination of size and size distribution of carbon black particles. Sedimentation and flow field-flow fractionations (FIFFF) were used to determine the size of carbon black, and their optimum analytical conditions were tested by changing surfactant, pH, ionic strength, and method of dispersion. The results showed that surfactant structure and its concentration played significant roles in dispersion stability. Carbon black was dispersed well with a nonionic surfactant with a pH of around 8 and an ionic strength of 0.003 M. The mean diameters measured from two types of FFF and photon correlation spectroscopy are in good agreement. This study demonstrates the potential of sedimentation and flow FFF for analyzing highly adsorptive industrial particles and guides for sample preparation.

Comparison of separation conditions and ionization methods for the liquid chromatography-mass spectrometric determination of sulfonamides.

The effects of the type and concentration of buffer, composition of the mobile phase and the ionization mode, used for the separation and detection of sulfonamides with LC-MS, were studied. Five typical sulfonamides were selected as target compounds and beef meat was selected as a matrix sample. For the separation of sulfonamides, 0.05 M NH4Ac in 13-15% aqueous acetonitrile, APCl ionization was more effective than ESI with regard to separation efficiency and the detection sensitivity.

Probing lysine acetylation with a modification-specific marker ion using high-performance liquid chromatography/electrospray-mass spectrometry with collision-induced dissociation.

Posttranslational acetylation of proteins regulates many diverse functions, including DNA recognition, protein-protein interaction, and protein stability. The identification of enzymes that regulate protein acetylation has revealed broader use of this modification than was previously suspected. In this study, we describe a method for identifying protein acetylation at lysine residues by analysis of digested protein using HPLC/ESI-MS with a new modification-specific marker ion. Collision-induced dissociation with capillary or nano-LC/ESI-TOF-MS was used to obtain a fragment ion useful as a marker for acetylated lysine. Although the acetylated lysine immonium ion at m/z 143.1 has been used as a marker ion for detecting acetylated lysine, it can be confused with internal fragment ion in some peptides, producing false positive results. We have found a novel marker ion at m/z 126.1, which is a further fragment ion induced by the loss of NH3 from the acetylated lysine immonium ions at m/z 143.1. This novel marker ion was found to be more specific and approximately 9 times more sensitive than the immonium ion at m/z 143.1. In addition, no interfering ions for acetylated peptides were found in the extracted ion chromatogram at m/z 126.1. The utility of this method was demonstrated with acetylated cytochrome c as a model compound. After the modification was probed by the new marker ion, the acetylated lysine site was determined by the CID-MS spectrum. This method was applied to identify histone H4 acetylation in HeLa cells treated with trichostatin A. Three protein bands separated by acid-urea-Triton gel electrophoresis were confirmed as tetra, tri, and diacetylated histone H4 at lysines 5, 8, 12, and 16. This method may be useful for assaying for lysine acetylation, which is an important regulatory process for a range of biological functions.

Size characterization of incinerator fly ash using sedimentation/steric field-flow fractionation.

Fly ash particles emitted from municipal solid waste-incinerators are of environmental concern. This study aims to investigate the applicability of sedimentation/steric field-flow fractionation (Sd/StFFF) and to develop a Sd/StFFF method for the separation and size characterization of incinerator fly ash. This study focuses on the fly ash particles larger than approxiamtely 1 microm, which comprise more than 90% (w/w) of the fly ash. Fly ash is a complex mixture of particles having various chemical compositions, sizes, shapes, and densities. Prior to Sd/StFFF analysis, fly ash particles are prefractionated into six density classes using a modified centrifugal procedure. It was found that fly ash particles are most abundant in the density range between 2.4 and 2.8 g/cm3. Different density fractions seem to contain particles of different chemical compositions. The Sd/StFFF conditions for the size-characterization of fly ash are sample concentration, approximately 0.3% (w/v); dispersing medium, 50% ethanol in water; and carrier liquid, water with 1.0% FL-70 (ionic strength approximately 0.012 M). Sd/StFFF data show no significant differences in size distribution among different density fractions. Generally, the sizes obtained from Sd/StFFF are larger than those obtained from a Coulter Multisizer and microscopy, probably because of the irregular shapes of the fly ash particles.