Home About us Contact
|
Home About us Contact | ||
|
|
||
Fractionation
Kinds of Fractionation Terms modified by Fractionation Selected AbstractsDIET-TISSUE FRACTIONATION OF STABLE CARBON AND NITROGEN ISOTOPES IN PHOCID SEALSMARINE MAMMAL SCIENCE, Issue 1 2002VÉRONIQUE Lesage Diet-tissue isotopic fractionation of carbon (C) and nitrogen (N) isotopes in short- and longer-term diet integrators of diet (i. e., blood serum and red cells), that involve non-invasive sampling techniques was examined using three species of phocid seals (harbor seals, gray seals, and harp seals) fed a known diet. Variability in diet-tissue fractionation values within and between species was also scrutinized to determine the legitimacy of using values obtained from one species to explore trophic positions and diets of other related species. All captive seals raised on a constant diet had tissues enriched in 13C and 15N relative to their diet. Diet-tissue isotopic fractionation values were generally consistent among conspecifics and among phocid species for a given tissue. Trophic isotopic enrichment in 13C was significantly higher in red blood cells (+1.5%±) than in blood serum (+0.8%±), whereas the reverse was observed for nitrogen isotopes (+1.7%± in red cells vs. +3.1%± in serum). However, 13C-depleted lipids were not extracted from blood tissues in this study. This results in a downward bias in the diet-tissue fractionation factors for carbon for both red cells and blood serum, particularly the latter because of their significantly higher lipid contents (x,± SD = 14.6 ± 2.3%; n= 20; red blood cells 3.8 ± 0.9%±; n= 50, muscle 7.7 ± 2.0; n= 21) in marine mammals. [source] Protein methylation in full length Chlamydomonas flagellaCYTOSKELETON, Issue 8 2009Roger D. Sloboda Abstract Post-translational protein modification occurs extensively in eukaryotic flagella. Here we examine protein methylation, a protein modification that has only recently been reported to occur in flagella [Schneider MJ, Ulland M, Sloboda RD.2008. Mol Biol Cell 19(10):4319,4327.]. The cobalamin (vitamin B12) independent form of the enzyme methionine synthase (MetE), which catalyzes the final step in methionine production, is localized to flagella. Here we demonstrate, using immunogold scanning electron microscopy, that MetE is bound to the outer doublets of the flagellum. Methionine can be converted to S-adenosyl methionine, which then serves as the methyl donor for protein methylation reactions. Using antibodies that recognize symmetrically or asymmetrically methylated arginine residues, we identify three highly methylated proteins in intact flagella: two symmetrically methylated proteins of about 30 and 40 kDa, and one asymmetrically methylated protein of about 75 kDa. Several other relatively less methylated proteins could also be detected. Fractionation and immunoblot analysis shows that these proteins are components of the flagellar axoneme. Immunogold thin section electron microscopy indicates that the symmetrically methylated proteins are located in the central region of the axoneme, perhaps as components of the central pair complex and the radial spokes, while the asymmetrically methylated proteins are associated with the outer doublets. Cell Motil. Cytoskeleton 2009. © 2009 Wiley-Liss, Inc. [source] Bonding Form Analysis of Metals and Sulfur Fractionation in Methanol-Grown Anaerobic Granular SludgeENGINEERING IN LIFE SCIENCES (ELECTRONIC), Issue 5 2007A. van der Veen Abstract This study investigates the metal and sulfur bonding form distribution in mesophilic (30,°C, pH 7) methanol-grown anaerobic granular sludge from upflow anaerobic sludge bed reactors operating at an organic loading rate of 3.8,g CH3OH-COD/L d. This was achieved by applying a modified Tessier sequential extraction scheme to investigate the metal bonding forms and a sequential extraction scheme for sulfur and simultaneously extracted metals to granular sludge samples of the reactors after 0, 22, 35 and 43 days of operation. Metals were also determined in the sulfur extracts. Co and Ni predominated in their oxidizable bonding forms, which increased together with the pseudo-total content during reactor operation. An omission of Co and Ni from the influent led to only a minor decline of the pseudo-total content in the sludge, mainly from the acid-soluble fraction. The ratio of simultaneously extracted metals (Co, Fe, Mn, Ni) to acid-volatile sulfides was lower than 1, indicating that the sludge contained sufficient sulfide to bind the metals as metal monosulfides. The bioavailability of metals in the methanol-grown anaerobic granular sludge investigated is therefore mainly controlled by sulfide formation/dissolution. [source] Evaluation of the ishikawa cell line bioassay for the detection of estrogenic substances from sediment extractsENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 7 2005Shinya Hashimoto Abstract This study examines the application of Ishikawa human endometrial adenocarcinoma cells to measure the estrogenic activity of fractionated extracts of sediments from Tokyo Bay, Japan. Estrogen stimulates alkaline phosphatase activity in this cell line. The results of these assays were compared with those of a yeast estrogen screen (YES) assay. The Ishikawa cell line bioassay showed higher sensitivity to 17,-estradiol (median effective concentration [EC50], 10.7 pM) than did the YES assay (EC50, 480 pM). Fractionation of sediment extracts (all samples collected from 5 sites) showed that the nonpolar fraction was poisonous to yeast cells; the estrogenic activity of this fraction, therefore, could not be measured by YES. However, the nonpolar fraction did not kill the Ishikawa cells. The 17,-estradiol-equivalent values of 15 extracts (3 fractions from each of 5 sediment samples) ranged from 5.7 to 697 pg/g dry weight according to the Ishikawa cell line bioassay. Chemical analysis using gas chromatography-mass spectrometry revealed that the highest concentrations of endocrine-disrupting chemicals were observed at the sampling station near the sewage treatment plant. The results support that the Ishikawa cell line bioassay is suitable for measuring the estrogenic activity of sediment samples. [source] BJ46a, a snake venom metalloproteinase inhibitorFEBS JOURNAL, Issue 10 2001Isolation, characterization, cloning, insights into its mechanism of action Fractionation of the serum of the venomous snake Bothrops jararaca with (NH4)2SO4, followed by phenyl-Sepharose and C4 -reversed phase chromatographies, resulted in the isolation of the anti-hemorrhagic factor BJ46a. BJ46a is a potent inhibitor of the SVMPs atrolysin C (class P-I) and jararhagin (P-III) proteolytic activities and B. jararaca venom hemorrhagic activity. The single-chain, acidic (pI 4.55) glycoprotein has a molecular mass of 46 101 atomic mass units determined by MALDI-TOF MS and 79 kDa by gel filtration and dynamic laser light scattering, suggesting a homodimeric structure. mRNA was isolated from the liver of one specimen and transcribed into cDNA. The cDNA pool was amplified by PCR, cloned into a specific vector and used to transform competent cells. Clones containing the complete coding sequence for BJ46a were isolated. The deduced protein sequence was in complete agreement with peptide sequences obtained by Edman degradation. BJ46a is a 322-amino-acid protein containing four putative N-glycosylation sites. It is homologous to the proteinase inhibitor HSF (member of the fetuin family, cystatin superfamily) isolated from the serum of the snake Trimeresurus flavoviridis, having 85% sequence identity. This is the first report of a complete cDNA sequence for an endogenous inhibitor of snake venom metalloproteinases (SVMPs). The sequence reveals that the only proteolytic processing required to obtain the mature protein is the cleavage of the signal peptide. Gel filtration analyses of the inhibitory complexes indicate that inhibition occurs by formation of a noncovalent complex between BJ46a and the proteinases at their metalloproteinase domains. Furthermore, the data shows that the stoichiometry involved in this interaction is of one inhibitor monomer to two enzyme molecules, suggesting an interesting mechanism of metalloproteinase inhibition. [source] Identification of phospholipids as new components that assist in the in vitro trimerization of a bacterial pore proteinFEBS JOURNAL, Issue 3 2001Hans De Cock The in vitro trimerization of folded monomers of the bacterial pore protein PhoE, into its native-like, heat- and SDS-stable form requires incubations with isolated cell envelopes and Triton X-100. The possibility that membranes could be isolated that are enriched in assembly factors required for assembly of the pore protein was now investigated. Fractionation of total cell envelopes of Escherichia coli via various techniques indeed revealed the existence of membrane fractions with different capacities to support assembly in vitro. Fractions containing mainly inner membrane vesicles supported the formation of trimers that were associated with these membrane vesicles. However, only a proportion of these trimers were heat- and SDS-stable and these were formed with slow kinetics. In contrast, fractions containing mainly outer membrane vesicles supported formation of high amounts of heat-stable trimers with fast kinetics. We identified phospholipids as active assembly components in these membranes that support trimerization of folded monomers in a process with similar characteristics as observed with inner membrane vesicles. Furthermore, phospholipids strongly stimulate the kinetics of trimerization and increase the final yield of heat-stable trimers in the context of outer membranes. We propose that lipopolysaccharides stabilize the assembly competent state of folded monomers as a lipochaperone. Phospholipids are involved in converting the folded monomer into new assembly competent intermediate with a short half-life that will form heat-stable trimers most efficiently in the context of outer membrane vesicles. These results provide biochemical evidence for the involvement of different lipidic components at distinct stages of the porin assembly process. [source] Factors controlling the carbon isotope fractionation of tetra- and trichloroethene during reductive dechlorination by Sulfurospirillum ssp. and Desulfitobacterium sp. strain PCE-SFEMS MICROBIOLOGY ECOLOGY, Issue 1 2007Danuta Cichocka Abstract Carbon stable isotope fractionation of tetrachloroethene (PCE) and trichloroethene (TCE) was investigated during reductive dechlorination. Growing cells of Sulfurospirillum multivorans, Sulfurospirillum halorespirans, or Desulfitobacterium sp. strain PCE-S, the respective crude extracts and the abiotic reaction with cyanocobalamin (vitamin B12) were used. Fractionation of TCE (,C=1.0132,1.0187) by S. multivorans was more than one order of magnitude higher than values previously observed for tetrachloroethene (PCE) (,C=1.00042,1.0017). Similar differences in fractionation were observed during reductive dehalogenation by the close relative S. halorespirans with ,C=1.0046,1.032 and ,C=1.0187,1.0229 for PCE and TCE respectively. TCE carbon isotope fractionation (,C=1.0150) by the purified PCE-reductive dehalogenase from S. multivorans was more than one order of magnitude higher than fractionation of PCE (,C=1.0017). Carbon isotope fractionation of TCE by Desulfitobacterium sp. strain PCE-S (,C=1.0109,1.0122) as well as during the abiotic reaction with cyanocobalamin (,C=1.0154) was in a similar range to previously reported values for fractionation by mixed microbial cultures. In contrast with previous results with PCE, no effects due to rate limitations, uptake or transport of the substrate to the reactive site could be observed during TCE dechlorination. Our results show that prior to a mechanistic interpretation of stable isotope fractionation factors it has to be carefully verified how other factors such as uptake or transport affect the isotope fractionation during degradation experiments with microbial cultures. [source] Composition-Induced Variations in SIMS Instrumental Mass Fractionation during Boron Isotope Ratio Measurements of Silicate GlassesGEOSTANDARDS & GEOANALYTICAL RESEARCH, Issue 1 2008Martin Rosner isotopes du bore; SIMS; effet de matrice; matériaux vitreux de référence; fractionnement de masse instrumental An analytical artefact is reported here related to differences in instrumental mass fractionation between NIST SRM glasses and natural geological glasses during SIMS boron isotope determinations. The data presented demonstrated an average 3.4, difference between the NIST glasses and natural basaltic to rhyolitic glasses mainly in terms of their sputtering-induced fractionation of boron isotopes. As no matrix effect was found among basaltic to rhyolitic glasses, instrumental mass fractionation of most natural glass samples can be corrected by using appropriate glass reference materials. In order to confirm the existence of the compositionally induced variations in boron SIMS instrumental mass bias, the observed offset in SIMS instrumental mass bias has been independently reproduced in two laboratories and the phenomenon has been found to be stable over a period of more than one year. This study highlights the need for a close match between the chemical composition of the reference material and the samples being investigated. Nous montrons l'existence d'un artefact analytique reliéà différents fractionnements de masse instrumentaux, observés sur les verres NIST SRM et des verres naturels durant des mesures des isotopes de bore par SIMS. Les données montrent une différence d'environ 3.4, entre les verres NIST et les verres naturels, de composition variant de basaltique à rhyolitique, en termes de fractionnement des isotopes du bore principalement induit par le phénomène de dispersion. Comme aucun effet de matrice n'a été observé entre les verres basaltiques et les verres rhyolitiques, le fractionnement de masse instrumental de la plupart des verres naturels peut être corrigé en utilisant des verres de références appropriés. Dans le but de confirmer l'existence de biais de masse liéà la composition lors de mesure du bore par SIMS, nous avons reproduit indépendamment le décalage observé entre deux laboratoires et ce phénomène s'est révélé stable sur une période de plus d'un an. Cette étude met en lumière le besoin d'ajuster précisément les compositions chimiques des matériaux de référence et des échantillons à analyser. [source] Length Fractionation of Carbon Nanotubes Using Centrifugation,ADVANCED MATERIALS, Issue 9 2008Jeffrey A. Fagan Scalable separation of single-walled carbon nanotubes (SWCNTs)by length and chirality is critical to the adaptation of these materials for applications. Ultracentrifugation of SWCNTs within a density gradient produces chiral separation of the NTs, and it is shown here that ultracentrifugation can also be used to produce length fractionated SWCNTs by exploiting their transient motion in response to applied centripetal acceleration. [source] Size Fractionation of Metal Nanoparticles by Membrane Filtration,ADVANCED MATERIALS, Issue 5 2005A. Akthakul A novel thin film composite nanofiltration (NF) membrane is fabricated by coating a conventional ultrafiltration membrane with a self-assembling amphiphilic graft copolymer. The NF membranes are used in the fractionation of gold nanoparticles to achieve a well-defined particle cutoff diameter and reduced size dispersity (see Figure). [source] Fractionation of Electrograms and Linking of Activation During Pharmacologic Cardioversion of Persistent Atrial Fibrillation in the GoatJOURNAL OF CARDIOVASCULAR ELECTROPHYSIOLOGY, Issue 5 2004ZHAOLIANG SHAN M.D. Introduction: During atrial fibrillation (AF), there is fractionation of extracellular potentials due to head-to-tail interaction and slow conduction of fibrillation waves. We hypothesized that slowing of the rate of AF by infusion of a Class I drug would increase the degree of organization of AF. Methods and Results: Seven goats were instrumented with 83 epicardial electrodes on the left atrium, left atrial appendage, Bachmann's bundle, right atrium, and right atrial appendage. AF was induced and maintained by an automatic atrial fibrillator. After AF had persisted for 4 weeks, the Class IC drug cibenzoline was infused at a rate of 0.1 mg/kg/min. AF cycle length (AFCL), the percentage of fractionated potentials, conduction velocity (CV), and direction of propagation of the fibrillation waves were measured during baseline, after AFCL was increased by 20, 40, 60, and 80 ms, and shortly before cardioversion. Infusion of cibenzoline increased the mean of the median AFCLs from 96 ± 6 ms to 207 ± 43 ms (P < 0.0001). The temporal variation in AFCL in different parts of the atria was 8% to 20% during control and, with the exception of Bachmann's bundle, was not significantly reduced during cibenzoline infusion. CV decreased from 76 ± 14 ms to 52 ± 9 cm/s (P < 0.01). Cibenzoline increased the percentage of single potentials from 81%± 4% to 91%± 4% (P < 0.01) and decreased the incidence of double potentials from 14%± 4% to 7 ± 5% (P < 0.01) and multiple potentials from 5%±% to 1%± 2% (P < 0.001). Whereas during control, linking (consecutive waves propagating in the same direction) during seven or more beats occurred in 9%± 15% of the cycles, after cibenzoline the degree of linking had increased to 40%± 33% (P < 0.05). During the last two beats before cardioversion, there was a sudden prolongation in AFCL from 209 ± 37 ms to 284 ± 92 ms (P < 0.01) and a strong reduction in fractionated potentials (from 22%± 12% to 6%± 5%, P < 0.05). Conclusion: The Class IC drug cibenzoline causes a decrease in fractionation of fibrillation electrograms and an increase in the degree of linking during AF. This supports the observation that Class I drugs widen the excitable gap during AF. (J Cardiovasc Electrophysiol, Vol. 15, pp. 572-580, May 2004) [source] Chemical Characteristics of Low-Fat Soymilk Prepared by Low-Speed Centrifugal Fractionation of the Raw SoymilkJOURNAL OF FOOD SCIENCE, Issue 5 2010Zhi-Sheng Liu Abstract:, Large oil,protein particles (2 to 60 ,m) were found in raw soymilk (or water extract of soybean), which was prepared in specific conditions. The large particles could be separated by sedimentation by centrifuging raw soymilk for 5 to 30 min at a low gravitational force ranging from 96 to 2410 ×,g. Chemical analysis showed that 80% to 90% of the total lipids and 30% to 40% of the total proteins were located in the precipitated fraction. The supernatant fraction had a dramatically higher protein-to-lipid ratio than the whole soymilk. The ratio of 11S/7S proteins and the ratio of 11S acidic/basic subunits were significantly (P,< 0.05) higher in the precipitate than that either in the whole soymilk or in the supernatant. Besides centrifuging conditions, other factors, including soymilk concentration, grinding method, soybean variety, and soybean storage, also significantly (P,< 0.05) affected the centrifugal fractionation. This study showed that low-speed centrifugation facilitated the separation of oil-protein particles from raw soymilk, and can be used as an innovative method for preparing low-fat soymilk and 11S protein-enriched ingredients. The findings also increased our understanding of the association or aggregation between proteins and lipids in raw soymilk after grinding. Practical Application:, Soymilk has become a popular beverage in the Western world due to its health benefits. Consumer demands for low-fat and organic foods have been increasing in the recent years. Currently, there are no alternative methods for manufacturing low-fat soymilk from whole soybeans. We found that most, if not all, of lipids in the raw soymilk were located in large particles, which could be separated by low-speed centrifugation. This centrifugal fractionation was investigated by varying processing parameters, soybean varieties, and soybean storage conditions. The approach has potential to be used for manufacturing low-fat soymilk. This study also has increased our understanding of the interactions between lipid and protein in raw soymilk. [source] Properties of High-Oleic Palm Oils Derived by Fractional CrystallizationJOURNAL OF FOOD SCIENCE, Issue 3 2008M.R. Ramli ABSTRACT:, High-oleic palm oil (HOPO) with an oleic acid content of 59.0% and an iodine value (IV) of 78.2 was crystallized in a 200-kg De Smet crystallizer with a predetermined cooling program and appropriate agitation. The slurry was then fractionated by means of dry fractionation at 4, 8, 10, 12, and 15 °C. The oil and the fractionated products were subjected to physical and chemical analyses, including fatty acid composition, triacylglycerol and diacylglycerol composition, solid fat content, cloud point, slip melting point, and cold stability test. Fractionation at 15 °C resulted in the highest olein yield but with minimal oleic acid content. Due to the enhanced unsaturation of the oil, fractionation at relatively lower crystallization temperature showed a considerable effect on fatty acid composition as well as triacylglycerol and diacylglycerol composition of liquid fractions compared to higher crystallization temperature. The olein and stearin fractionated at 4 °C had the best cold stability at 0 °C and sharper melting profile, respectively. [source] Effects of NaCl Concentration on Salting-in and Dilution During Salting-out on Soy Protein FractionationJOURNAL OF FOOD SCIENCE, Issue 4 2006N. A. Deak ABSTRACT:, Glycinin and ,-conglycinin are the main storage proteins in soybeans that can be fractionated by using alkali extraction, SO2, salting-in with NaCl, salting-out by dilution and pH adjustment to produce a glycinin-rich fraction, a ,-conglycinin,rich fraction, and an intermediate fraction, which is a mixture of the two proteins. Two different strategies were employed to optimize the procedure to achieve high efficiency in recovering the ,-conglycinin,rich fraction. The first strategy was to optimize salting-in effects of NaCl, and the effects of NaCl concentration on the yields and purities of the protein fractions were investigated. The maximum protein yield of the ,-conglycinin,rich fraction was obtained at 500 mM NaCl, but at the expense of purity. The optimum NaCl concentration was 250 mM, at which good protein yield (18.5%) and purity (84.5%) were achieved. At higher NaCl concentrations, the protein yields of the intermediate fractions were significantly lower, and the protein loss in the whey fraction increased. The second strategy was to improve the salting-out step for the ,-conglycinin,rich fraction. At 0- and 0.5-fold dilution, the purities and yields of the ,-conglycinin,rich fractions were significantly lower than at 1.0- and 2.0-fold dilution. There were no differences in protein yields or purities when using 1.0- or 2.0-fold dilution. According to these results, the recommended NaCl concentration for the salting-in step is 250 mM and the dilution factor for salting-out is 1.0. [source] Fractionation of Caseins by Anion-exchange Chromatography Using Food-grade BuffersJOURNAL OF FOOD SCIENCE, Issue 5 2003K.N. Turhan ABSTRACT : Caseins prepared by microfiltration of bovine skim milk were fractionated using anion-exchange chromatography. Laser densitometry of electrophoresis gels was shown to be sufficiently quantitative to perform accurate mass balance calculations detailing the fate of each casein fraction. L-cysteine was successfully used as a reducing agent instead of traditional toxic agents, such as dithiothreitol or ,-mercaptoethanol, enabling development of the first food-grade buffer system for casein fractionation. More salt was required for elution of the casein fractions having a greater charge: ,s -casein > ,-casein > ,-casein. Increasing flow rate decreased the extent of separation. Use of smaller beads was suggested as a method to maintain separation at increased flow rate. [source] TT virus (TTV) loads associated with different peripheral blood cell types and evidence for TTV replication in activated mononuclear cellsJOURNAL OF MEDICAL VIROLOGY, Issue 2 2001Fabrizio Maggi Abstract TT virus (TTV) loads associated with the peripheral blood cells of seven patients known to carry the virus in plasma were investigated by real-time PCR. Whereas red cells/platelets were uniformly negative, six and four patients yielded positive peripheral blood mononuclear cells (PBMCs) and polymorphonuclear leukocytes, respectively, but viral titres were generally low. Fractionation of PBMCs into monocyte- and B, T4, and T8 lymphocyte-enriched subpopulations showed no pattern in the viral loads that might suggest the preferential association of TTV to one or more specific cell types. TTV-negative PBMCs absorbed measurable amounts of virus when incubated with infected plasma at 4°C. Furthermore, cultures of TTV-negative phytohaemagglutinin-stimulated PBMCs exposed in vitro to virus-positive plasma and faecal extracts released considerable levels of infectious TTV into the supernatant fluid and the same was true for TTV-positive stimulated PBMCs. These results indicate that, whereas freshly harvested resting PBMCs seem to produce little, if any TTV, stimulated PBMCs actively replicate the virus. J. Med. Virol. 64:190,194, 2001. © 2001 Wiley-Liss, Inc. [source] An extract of Lannea microcarpa: composition, activity and evaluation of cutaneous irritation in cell cultures and reconstituted human epidermisJOURNAL OF PHARMACY AND PHARMACOLOGY: AN INTERNATI ONAL JOURNAL OF PHARMACEUTICAL SCIENCE, Issue 7 2006P. Picerno Lannea microcarpa (Anacardiaceae) is a tropical tree used in African folk medicine and commercial dermopharmaceutical formulations. Fractionation and analysis of its polar extract allowed the identification of 4,-methoxy-myricetin 3- O -,- l -rhamnopyranoside, myricetin 3- O -,- l -rhamnopyranoside, myricetin 3- O -,- d -glucopyranoside, vitexin, isovitexin, gallic acid and epi-catechin, as the major constituents. In-vivo assay (the croton oil ear test in mice) showed that the extract had significant anti-inflammatory effect (ID50 = 900 ,g cm2) but ten times lower than that of indometacin (ID50 = 93 ,g cm2), the non-steroidal anti-inflammatory drug used as reference. Cytotoxicity and cutaneous irritation of the extract and its constituents were investigated. The crude extract and its major components did not affect cell viability in-vitro either in three different cultures (J774.A1, WEHI-164 and HEK-293) of cells grown in monolayers or in the reconstituted human epidermis (RHE, 3D model), nor did they cause release of pro-inflammatory mediators (IL-1,) or histomorphological modification of RHE. [source] Book News: Microthermal Field-Flow Fractionation: Analysis of Synthetic, Natural, and Biological Macromolecules and Particles.JOURNAL OF SEPARATION SCIENCE, JSS, Issue 5-6 2009Edited by Josef Janca [source] Characterization via liquid chromatography coupled to diode array detector and tandem mass spectrometry of supercritical fluid antioxidant extracts of Spirulina platensis microalgaJOURNAL OF SEPARATION SCIENCE, JSS, Issue 9-10 2005Jose A. Mendiola Abstract Spirulina platensis microalga has been extracted on a pilot scale plant using supercritical fluid extraction (SFE) under various extraction conditions. The extraction yield and the antioxidant activity of the extracts were evaluated in order to select those extracts with both the highest antioxidant capacity and a good extraction yield. These extracts were characterized using LC coupled to diode array detection (DAD) and LC coupled to mass spectrometry (MS) with two different interfaces, atmospheric pressure chemical ionization (APCI) and electrospray (ESI) which allowed us to perform tandem MS by using an ion trap analyzer. The best extraction conditions were as follows: CO2 with 10% of modifier (ethanol) as extraction solvent, 55°C (extraction temperature) and 220 bar (extraction pressure). Fractionation was achieved by cascade depressurization providing two extracts with different activity and chemical composition. Several compounds have been identified in the extracts, corresponding to different carotenoids previously identified in Spirulina platensis microalga along with chlorophyll a and some degradation products. Also, the structure of some phenolic compounds could be tentatively identified. The antioxidant activity of the extracts could be attributed to some of the above mentioned compounds. [source] Sedimentation field-flow fractionation and granulometric analysis of PLGA microspheresJOURNAL OF SEPARATION SCIENCE, JSS, Issue 15-16 2003Nathalie Faisant Abstract Sedimentation field flow fractionation operated in the steric hyperlayer mode was used to obtain fractions of defined characteristics from crude samples of poly(D,L-lactic-co-glycolic acid) microspheres which were polydisperse in size. In less than ten minutes, Sedimentation Field Flow Fractionation (SdFFF) separation yielded three analytical fractions of very different size and particle size distribution (PSD) characteristics, as determined by granulometric analyses (Coulter Counter® and image analysis of SEM). A crude sample (average size = 45 ,m, 105% size polydispersity index) was separated into fractions of 73 ,m, 56 ,m, 8 ,m average diameters which showed a PSD of 39%, 33%, 30%, respectively. Our results demonstrated that SdFFF used in conjunction with particle size analysis offers a new approach to laboratory scale production of drug vectors of a specified average size and reduced size dispersity. In the future, this could be used to select the most convenient particles for drug loading and release. [source] Characterisation of the leaf meals, protein concentrates and residues from some tropical leguminous plantsJOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 9 2006J Oluwasola Agbede Abstract Leaf meals (LMs) from freshly harvested leaves of butterfly pea (Centrosema pubescens), devil bean (Mucuna pruriens), flamboyant flower (Delonix regia), Bauhinia tomentosa, coast wattle (Acacia auriculiformis), quick stick (Glyricidia sepium) and ipil-ipil (Leucaena leucocephala) were analysed for their nutrient and anti-nutritional content. Then, leaf protein concentrates (LPCs) were produced from the leaves by fractionation and characterised along with the fibrous residues. On average, the LM contained 181 g kg,1 dry matter (DM) CP (range: 100,280 g kg,1 DM), 139 g kg,1 DM crude fibre (range: 77,230 g kg,1 DM) and 133 g kg,1 DM ether extract (range: 86,165 g kg,1 DM) while the gross energy averaged 17.0 MJ kg,1. On average, leaf protein fractionation enhanced the CP, ether extract and the gross energy in the LPC by 39.5%, 33.5% and 22.0%, respectively, while the crude fibre of the LMs was reduced by 41%, on average, in the LPCs. Fractionation reduced the mineral content of the leaves generally. The mean phytin content varied from 0.36 g kg,1 in LPCs to 0.86 g kg,1 in leaf meal, while the mean phytin-P content varied from 0.10 g kg,1 in LPCs to 0.24 g kg,1 in leaf meal. The total phenol levels in the LMs were reduced by 33.7% in the LPCs, on average. These results suggest that, while the LPCs from these plants could be used as protein supplements in non-ruminant feeds in regions where there is an acute shortage of plant protein, the LMs or LPC fibrous residues could be fed to ruminant animals. Copyright © 2006 Society of Chemical Industry [source] Fractionation of Methyl Cellulose According to Polarity , a Tool to Differentiate First and Second Order Heterogeneity of the Substituent DistributionMACROMOLECULAR CHEMISTRY AND PHYSICS, Issue 11 2006Roland Adden Abstract Summary: A set of four MCs (DS 1.80,1.95) has been analyzed with regard to their substituent pattern in the glucosyl units and along the polymer chain. The average heterogeneity of methylation observed for the entire material was analyzed in more detail after fractionation according to polarity. All fractions obtained were analyzed with respect to their DS, monomer composition and deviation from a random distribution of these monomers in the polymer chains. By this approach, heterogeneity of first and second order could be differentiated. While for three of the MCs only a minor DS-gradient over the material was observed, a more pronounced heterogeneity of first order was obtained for MC 4. ESI mass spectrum of the undissolved residue of MC 2 after deuteromethylation and partial hydrolysis; DP 2 and 3 are shown in detail. Signals are assigned according to the number of CH3 -groups. [source] Effects of the Reaction Conditions on the Syndiospecific Polymerization of Propene Promoted by Bis(phenoxyimine) Titanium CatalystsMACROMOLECULAR CHEMISTRY AND PHYSICS, Issue 4 2004Marina Lamberti Abstract Summary: The propene polymerization behavior of two typical bis(phenoxyimine) titanium catalysts has been investigated by varying reaction conditions, such as the monomer concentration, the solvent, and the cocatalyst. The experimental results indicate that the stereoregularity and regioregularity of the obtained poly(propylene)s are significantly affected by the reaction conditions. Fractionation of some poly(propylene) samples indicates the formation of macromolecules of different stereoregularity in the same run, suggesting that different active complexes can be generated in situ from these bis(phenoxyimine) titanium precatalysts. [source] Simulation of Crystallization Analysis Fractionation (Crystaf) of Linear Olefin Block CopolymersMACROMOLECULAR SYMPOSIA, Issue 1 2009Siripon Anantawaraskul Abstract Summary: Linear olefin block copolymers (OBCs) have microstructures that are unique among polyolefins and exhibit properties that are different from those of other polyolefin elastomers. Characterizing their chain microstructures is a challenging task, as conventional characterization techniques cannot probe directly block length distribution or composition. In this work, we used a Monte Carlo model to predict the microstructure details of OBCs and a modified version of the Crystaf model previously developed in our groups to describe theoretical Crystaf profiles for model OBCs. This model can be used as a tool to interpret Crystaf results of these interesting new polyolefins and to relate them to OBC microstructures. Effects of polymerization parameters on OBC microstructure and Crystaf profiles were also discussed. [source] Metabolism of curcumin and induction of mitotic catastrophe in human cancer cellsMOLECULAR NUTRITION & FOOD RESEARCH (FORMERLY NAHRUNG/FOOD), Issue 9 2008Julia S. Dempe Abstract In cultured cells, curcumin (CUR) causes cell death by interfering with mitosis and leading to fragmented nuclei and disrupted microtubules, a process named mitotic catastrophe. In order to clarify the role of the known CUR metabolites hexahydro-CUR (HHC) and CUR-glucuronide (CUR-gluc) in mitotic catastrophe, the effects of CUR were studied in three human cancer cell lines with different metabolism of CUR. In Ishikawa and HepG2 cells, CUR was metabolized to HHC and small amounts of octahydro-CUR (OHC), whereas the only metabolism in HT29 cells was the formation of CUR-gluc. Despite their different metabolism, all three cell systems responded to CUR with arrest in G2/M phase and mitotic catastrophe. Fractionation of the cells showed that concentrations of CUR were higher in the ER and cytosol than in the incubation medium by a factor of up to about 150 and 8, respectively. In contrast to CUR, the metabolite HHC and the products of spontaneous degradation did not elicit any effects in Ishikawa cells. These results imply that the causative agent of mitotic catastrophe is the parent CUR molecule, whereas reductive metabolism and chemical degradation render CUR inactive. [source] Chemistry and genotoxicity of caramelized sucroseMOLECULAR NUTRITION & FOOD RESEARCH (FORMERLY NAHRUNG/FOOD), Issue 12 2006David D. Kitts Abstract Caramelization of a 1% sucrose solution at 180°C accompanied characteristic changes in pH, Mr, UV-absorbance, and fluorescence values as well as increased reducing power activity after 40,60 min. Similar changes occurred to sucrose heated at 150°C, after 150,240 min. Bioactivity of caramelized sucrose samples was tested for mutagenic activity, using Salmonella typhimurium strains TA-98 and TA-100, respectively, as well as the Saccharomyces D7 yeast strain for mitotic recombination and Chinese hamster ovary cells (CHO) to assess clastogenicity. Caramelized sucrose expressed no mutagenicity in the TA-98 strain, but gave positive (p < 0.05) results with the TA-100, base-pair substitution strain. Similarly, mitotic recombination in the Saccharomyces D7 yeast strain and clastogenic activity in CHO cells were induced when exposed to caramelized sucrose. In the all cases, preincubation with S-9 reduced (p < 0.05) the mutagenic activities of caramelized sucrose. Fractionation of the caramelized sucrose into volatile and nonvolatile compounds was performed and tested for clastogenicity using CHO cells. Volatile components contributed approximately 10% to total clastogenicity, which was enhanced by the presence of S-9. Nonvolatile components recovered, consisting of relatively lower Mr, gave highest (p < 0.05) clastogenic activity, denoting that higher Mr caramel colors are relatively free of this property. [source] Hypericin-mediated Photocytotoxic Effect on HT-29 Adenocarcinoma Cells Is Reduced by Light Fractionation with Longer Dark Pause Between Two Unequal Light DosesPHOTOCHEMISTRY & PHOTOBIOLOGY, Issue 6 2005Veronika Sa ABSTRACT The present study demonstrates the in vitro effect of hypericin-mediated PDT with fractionated light delivery. Cells were photosensitized with unequal light fractions separated by dark intervals (1 or 6 h). We compared the changes in viability, cell number, survival, apoptosis and cell cycle on HT-29 cells irradiated with a single light dose (12 J/cm2) to the fractionated light delivery (1 + 11 J/cm2) 24 and 48 h after photodynamic treatment. We found that a fractionated light regime with a longer dark period resulted in a decrease of hypericin cytotoxicity. Both cell number and survival were higher after light sensitization with a 6-h dark interval. DNA fragmentation occurred after a single light-dose application, but in contrast no apoptotic DNA formation was detected with a 6-h dark pause. After fractionation the percentage of cells in the G1 phase of the cell cycle was increased, while the proportion of cells in the G2 phase decreased as compared to a single light-dose application, i.e. both percentage of cells in the G1 and G2 phase of the cell cycle were near control levels. We presume that the longer dark interval after the irradiation of cells by first light dose makes them resistant to the effect of the second illumination. These findings confirm that the light application scheme together with other photodynamic protocol components is crucial for the photocytotoxicity of hypericin. [source] Fractionation of grape tannins and analysis by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometryPHYTOCHEMICAL ANALYSIS, Issue 4 2003Camille Perret Abstract Polymeric tannins, extracted from grape berries (Gamay variety), were fractionated according to their mean degree of polymerisation (mDP) on a styrene,divinylbenzene phase eluted with a gradient of methanol:chloroform. Increasing the percentage of methanol led to the solubilisation of higher molecular weight tannins. The mean mDP of each collected fraction was determined by acid-catalysed degradation in the presence of a nucleophilic reagent. The fractionation method produced a linear gradient of mDP varying between 1.84 and 19.34. The fractions were partially characterised by matrix assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF/MS). The spectra showed a complex mixture of proanthocyanidins and galloylated proanthocyanidins up to 4000,amu. Copyright © 2003 John Wiley & Sons, Ltd. [source] Sequential detergent fractionation of primary neurons for proteomics studiesPROTEINS: STRUCTURE, FUNCTION AND BIOINFORMATICS, Issue 5 2008Simonetta Bernocco Dr. Abstract Proteomics studies employing primary neurons are difficult due to the neurons' characteristics. We have developed a detergent-based fractionation method which reduces complexity of the protein extracts, is sufficiently fast to allow differential proteomics analysis after treatments of neurons for short time periods, can be applied to small numbers of cells directly in culture plates, and allows differential extraction of proteins in a compartment-specific manner. The sequential use of detergent-containing buffers on neurons in culture plates yields four extracts enriched in cytosolic, membrane-bound or enclosed, nuclear, and cytoskeletal proteins. Fractionation of neurons was validated by comparison of the distribution of known subcellular marker proteins in the four extracts using Western blotting. Comparison of extracts by DIGE showed a clear difference in protein composition demonstrating significant variations with a fold change (FC) of at least 1.20 for 82% of the detected spots. Using proteins identified in these spots that could be assigned a subcellular localization based on descriptions in the Uniprot database, an extraction efficiency of 85% was calculated for cytosolic proteins in extract 1, 90% for membrane-bound and membrane-enclosed proteins in extract 2, 82% for nuclear proteins in extract 3 and 38% for cytoskeletal and RAFT proteins in extract 4. [source] Characterisation of organellar proteomes: A guide to subcellular proteomic fractionation and analysisPROTEINS: STRUCTURE, FUNCTION AND BIOINFORMATICS, Issue 21 2006Edwin Ho Abstract Subcellular fractionation is being widely used to increase our understanding of the proteome. Fractionation is often coupled with 2-DE, thus allowing the visualisation of proteins and their subsequent identification and characterisation by MS. Whilst this strategy should be effective, to date, there has been little or no consideration given to differences in the mass, pI, hydropathy or abundance of proteins in the organelles and how analytical strategies can be tailored to match the idiosyncrasies of proteins in each particular compartment. To address this, we analysed 3962 Saccharomyces cerevisiae proteins, previously localised to one or more of 22 subcellular compartments. Different compartments showed significantly different distributions of protein pI and hydropathy. Mitochondrial and ER proteins showed the most dramatic differences to other organelles, in their protein pIs and hydropathy, respectively. We show that organelles can be clustered by similarities in these physicochemical protein characteristics. Interestingly, the distribution of protein abundance was also significantly different between many organelles. Our results show that to fully explore subcellular fractions of the proteome, specific analytical strategies should be employed. We outline strategies for all 22 subcellular compartments. [source] | ||||||||||||||||||||||||||||||||||||||||||||||