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Heterogeneous Mixture (heterogeneous + mixture)
Selected AbstractsRecombinant clotting factor VIII concentrates: Heterogeneity and high-purity evaluationELECTROPHORESIS, Issue 16 2010Gian Maria D'Amici Abstract Factor VIII is an important glycoprotein involved in hemostasis. Insertion of expression vectors containing either the full-length cDNA sequence of human factor VIII (FLrFVIII) or B-domain deleted (BDDrFVIII) into mammalian cell lines results in the production of recombinant factor VIII (rFVIII) for therapeutic usage. Three commercially available rFVIII concentrates (Advate®, Helixate NexGen® and Refacto®), either FLrFVIII or BDDrFVIII, were investigated by 1- and 2-DE and MS. The objective of this study was to compare the heterogeneity and the high purity of both rFVIII preparations before and after thrombin digestion. In particular, the 2-D gel was optimized to better highlight the presence of contaminants and many unexpected proteins. Recombinant strategies consisting of insertion of expression vectors containing BDDrFVIII and FLrFVIII resulted in homogeneous and heterogeneous protein products, respectively, the latter consisting in a heterogeneous mixture of various B-domain-truncated forms of the molecule. Thrombin digestion of all the three rFVIII gave similar final products, plus one unexpected fragment of A2 domain missing 11 amino acids. Regarding the contaminants, Helixate NexGen® showed the presence of impurities, such as Hsp70,kDa, haptoglobin and proapolipoprotein; Refacto® showed glutathione S -transferase and ,-lactamase, whereas Advate® apparently did not contain any contaminants. The proteomic approach will contribute to improving the quality assurance and manufacturing processes of rFVIII concentrates. In this view, the 2-DE is mandatory for revealing the presence of contaminants. [source] Mass spectrometry study of ecto-5,-nucleotidase from bull seminal plasmaFEBS JOURNAL, Issue 16 2000Carlo Fini The structure of ecto-5,-nucleotidase from bull seminal plasma, containing a glycosyl-phosphatidylinositol anchor, was studied using mass spectrometry. MALDI-MS analysis of intact protein indicated a mass of 65 568.2 Da for the monomeric form, and it also showed a heterogeneous population of glycoforms with the glycosidic moiety accounting for ,,6000 Da. MALDI-MS analysis showed that Asn53, Asn311, Asn333 and Asn403 were four sites of N -glycosylation. GC-MS analysis provided information on the glycosidic structures linked to the four asparagines. Asn53, Asn311 and Asn333 were linked to high-mannose saccharide chains, whereas the glycan chains linked to Asn403 contained a heterogeneous mixture of oligosaccharides, the high-mannose type structure being the most abundant and hybrid or complex type glycans being minor components. By combining enzymatic and/or chemical hydrolysis with GC-MS analysis, detailed characterization of the glycosyl-phpsphatidylinositol anchor was obtained. MALDI spectral analysis indicated that the glycosyl-phosphatidylinositol core contained EtN(P)Man3GlcNH2 -myo-inositol(P)-glycerol, principally modified by stearoyl and palmitoyl residues or by stearoyl and myristoyl residues to a minor extent. Moreover, 1-palmitoylglycerol and 1-stearoylglycerol outweighed 2-palmitoylglycerol and 2-stearoylglycerol. The combination of chemical and enzymatic digestions of the protein with the mass spectral analysis yielded a complete pattern of S,S bridges. The protein does not contain free thiols and its eight cysteines are linked by intramolecular disulfide bonds, the pairs being: Cys51,Cys57, Cys353,Cys358, Cys365,Cys387 and Cys476,Cys479. This work resolves details of the structure of ecto-5,-nucleotidase, with particular regard to the localization and composition of the glycidic moiety, number and localization of the disulfide bridges and characterization of the glycosyl-phosphatidylinositol anchor. [source] Eco-friendly methodologies for the synthesis of some aromatic esters, well-known cosmetic ingredientsINTERNATIONAL JOURNAL OF COSMETIC SCIENCE, Issue 1 2005C. Villa Synopsis Solid,liquid solvent-free phase transfer catalysis (PTC) and acidic catalysis in dry media were applied, with noticeable improvement and simplification over classical procedures in a Green Chemistry context, to the synthesis of some aromatic esters useful as cosmetic ingredients: 3-methylbutyl 4-methoxycinnamate, 2-ethylhexyl 4-methoxycinnamate, 2-ethylhexyl 4-(dimethylamino)benzoate and 2-ethylhexyl salicylate, well-known ultraviolet B sunscreen filters; 4-isopropylbenzyl salicylate, UV absorber and cutaneous antilipoperoxidant; propyl 4-hydroxybenzoate and butyl 4-hydroxybenzoate (parabens), antimicrobial agents. The reactions were performed under microwave (MW) activation and conventional heating. The best results for the synthesis of cinnamic, salicylic and 4-(dimethylamino)benzoic esters were achieved by in situ preformed carboxylates alkylation with alkyl bromides using PTC. The 4-hydroxybenzoates were obtained in good yields by classical esterification of the acid with alcohols using a simple heterogeneous mixture of reagents with catalytic amounts of p -toluenesulfonic acid (PTSA). The comparisons of yields and thermal profiles under either MW or conventional heating were studied and reported. Résumé La catalyse par transfert de phase (CTP) solide-liquide sans solvant et l'estérification acido-catalysée en ,,milieu sec'' ont été appliquées, dans le cadre de la ,,chimie verte'', avec des nettes améliorations et simplifications par rapport aux méthodes classiques, à la synthèse de certains esters aromatiques, ingrédients cosmétiques: 3-methylbutyl 4-méthoxycinnamate, 2-éthylhéxyl 4-méthoxycinnamate, 2-éthylhéxyl 4-(diméthylamino) benzoate et 2-éthylhéxyl salicylate, filtres solaires UVB bien connus; 4-isopropylbenzyl salicylate, filtre UV et antilipopéroxydant cutané; propyl 4-hydroxybenzoate et butyl 4-hydroxybenzoate (parabens), agents antimicrobien. Les réactions ont été procédées sous irradiation micro-onde et par chauffage classique. Pour la synthèse des esters dérivés de l'acide cinnamique, salicylique et 4-(diméthylamino)benzoïque les meilleurs résultats ont été obtenus grâce à la CTP par alkylation des carboxylates (préformés in situ) avec des bromures d'alkyle. Les esters de l'acide 4-hydroxybenzoique ont été obtenus avec de bons rendements par simple mélange hétérogène des réactifs et d'acide p-toluènesulfonique (PTSA) comme catalyseur. Les rendements et les profils de montée en température sous micro-ondes et par chauffage classique ont été comparés. [source] Green chemistry procedure for the synthesis of cyclic ketals from 2-adamantanone as potential cosmetic odourantsINTERNATIONAL JOURNAL OF COSMETIC SCIENCE, Issue 5 2002M. T. Genta Synopsis Some cyclic ketals derived from 2-adamantanone were obtained in excellent yields by microwave activation under solvent-free conditions, as a ,green chemistry' procedure. A number of experiments were performed to evaluate the most efficient catalytic conditions. The best results were obtained using a simple heterogeneous mixture of reagents and p -toluenesulphonic acid as the catalyst, without any solvent or support. The data are reported and compared with those obtained by other microwave-mediated syntheses or by classical method. In order to check the possible intervention of non-thermal microwave effects, the best experiment in ,dry media' was carried out with considerable lower yield by conventional heating, in a thermostated oil bath, under the same conditions as under microwaves (time, temperature and vessel). All the synthesized compounds were tested for their olfactive character and for a potential cosmetic use. The odour evaluation is reported. Résumé Des acétals cycliques dérivés de la 2-adamantanone ont été obtenus sous irradiation microonde sans solvant avec d'excellents rendements selon un procédé de ,chimie verte'. Un certain nombre d'experiences ont été réalisées pour optimiser les conditions de réaction. Les résultats obtenus par simple mélange hétérogène des réactifs et de l'acide p -toluènesulphonique 10% (p/p) comme catalyseur ont été décrits et comparés avec ceux obtenus avec les autres procédés, sous irradiation microonde ou par chauffage traditionnel. Dans le but de mettre en évidence l'éventuelle intervention d'effets spécifiques (non-purement thermiques) des microondes, la meilleure réaction obtenue en ,milieu sec' a été effectuée, avec des rendements nettement plus faibles, dans les mêmes conditions (temps et température), par chauffage classique. Les propriétés olfactives de tous les produits obtenus ont été déterminées pour évaluer une possible utilization cosmétique. [source] Two iridovirus-susceptible cell lines established from kidney and liver of grouper, Epinephelus awoara (Temminck & Schlegel), and partial characterization of grouper iridovirusJOURNAL OF FISH DISEASES, Issue 6 2000Y-S Lai Two iridovirus-susceptible cell lines were established and characterized from grouper Epinephelus awoara kidney and liver tissues. These cell lines have been designated GK and GL, respectively. The cells multiplied well in Leibovitz's L-15 medium, supplemented with 10% foetal bovine serum, at temperatures between 20 and 32 °C, and have been subcultured more than 120 times, becoming continuous cell lines. The cell lines consist of a heterogeneous mixture of fibroblastic and epithelial cells. The viability of cells, stored frozen in liquid nitrogen (,196 °C), was 95% after 1 year. Chromosome morphologies of GK and GL cells were homogeneous. Both cell lines were susceptible to grouper iridovirus, and yielded high titres of up to 108 TCID50 mL,1. In addition, both cell lines effectively replicated the virus, which could be purified to homogeneity by cesium chloride gradient centrifugation. Electron microscopy studies showed that purified virus particles were 170±10 nm in diameter, and were hexagonal in shape. Virus-infected cells showed an abundance of virus particles inside the cytoplasm. These results show that the GK and GL cell lines effectively replicate grouper iridovirus, and can be used as a tool for studying fish iridoviruses. [source] Reactions of Cl,/Cl2,, Radicals with the Nanoparticle Silica Surface and with Humic Acids: Model Reactions for the Aqueous Phase Chemistry of the AtmospherePHOTOCHEMISTRY & PHOTOBIOLOGY, Issue 4 2007Paula Caregnato ABSTRACT Reactions of chlorine radicals might play a role in aqueous aerosols where a core of inorganic components containing insulators such as SiO2 and dissolved HUmic-LIke Substances (HULIS) are present. Herein, we report conventional flash photolysis experiments performed to investigate the aqueous phase reactions of silica nanoparticles (NP) and humic acid (HA) with chlorine atoms, Cl,, and dichloride radical anions, Cl2,,. Silica NP and HA may be taken as rough models for the inorganic core and HULIS contained in atmospheric particles, respectively. Both Cl, and Cl2,, were observed to react with the deprotonated silanols on the NP surface with reaction rate constants, k ± ,, of (9 ± 6) × 107 M,1 s,1 and (7 ± 4) × 105 M,1 s,1, respectively. The reaction of Cl, with the surface deprotonated silanols leads to the formation of SiO, defects. HA are also observed to react with Cl, and Cl2,, radicals, with reaction rate constants at pH 4 of (3 ± 2) × 1010 M,1 s,1 and (1.2 ± 0.3) × 109 M,1 s,1, respectively. The high values observed for these constants were discussed in terms of the multifunctional heterogeneous mixture of organic molecules conforming HA. [source] SELDI-TOF MS analysis of the Cardiac Troponin,I forms present in plasma from patients with myocardial infarctionPROTEINS: STRUCTURE, FUNCTION AND BIOINFORMATICS, Issue 23 2006Estelle Peronnet Abstract The troponin,(Tn) complex is composed of troponin,T, troponin,C and troponin,I. The cardiac isoform of TnI (cTnI) is modified and released in blood of patients with cardiovascular diseases as a heterogeneous mixture of free, complexed and posttranslationally modified forms. With the aim to determine later, whether specific forms of cTnI could be associated with the different pathologies leading to cTnI release, the cTnI forms present in the plasma from 64,patients with acute myocardial infarction,(AMI) have been analysed by SELDI-TOF MS using anti-TnI mAbs coupled to PS20 ProteinChips® arrays. Upfront immunoaffinity enrichment using anti-cTnI,19C7 mAb allowed us to detect cTnI and bis -phosphorylated cTnI in 11/12 and 9/12 analyses respectively, as well as truncated cTnI in plasma with concentration of cTnI as low as 8,ng/mL. Cardiac troponin,C (cTnC) and covalent TnIC complex were also found in pools of plasma with higher concentrations of cTnI. MAb 19C7-affinity SELDI-TOF MS analysis performed after immunopurification of one pool of AMI plasma with anti-free cTnI, anti-cTnC, and anti-phosphorylated cTnI mAbs indicated that intact and bis -phosphorylated cTnI were mostly under the free form. Besides, a 18,718,m/z peak could correspond to a truncated phosphorylated form initially complexed with cTnC. [source] Elucidation of the molecular structure of lipid A isolated from both a rough mutant and a wild strain of Aeromonas salmonicida lipopolysaccharides using electrospray ionization quadrupole time-of-flight tandem mass spectrometryRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 12 2005Anas El-Aneed The chemical structure of lipid A, isolated by mild acid hydrolysis from a rough mutant and a wild strain of Aeromonas salmonicida lipopolysaccharide, was investigated using electrospray ionization quadrupole time-of-flight (QqToF) hybrid tandem mass spectrometry and showed a great degree of microheterogeneity. The chemical structure of the main constituent of this heterogeneous mixture was identified as a , -D-(1,,,6) linked D-glucosamine disaccharide substituted by two phosphate groups, one being bound to the non-reducing end at position O-4, and the other to the position O-1 of the reducing end of the D-glucosamine disaccharide. The location of the fatty acids linked to the disaccharide backbone was established by identifying diagnostic ions in the conventional QqToF-MS scan. Low-energy collision tandem mass spectrometry analysis of the selected precursor diagnostic ions confirmed, unambiguously, their proposed molecular structures. We have established that myristyloxylauric (C14:0(3- O(12:0))) acid residues were both N-2, and O-3, linked to the non-reducing end of the D-GlcN residue, and that two 3-hydroxymyristic (C14:0(3-OH)) acid chains acylated the remaining positions of the reducing end. The MS and MS/MS data obtained allowed us to determine the complex molecular structure of lipid A. The QqToF-MS/MS instrument has shown excellent superiority over a conventional quadrupole-hexapole-quadrupole tandem instrument which failed to fragment the selected precursor ion. Copyright © 2005 John Wiley & Sons, Ltd. [source] Back-scattered electron imaging and elemental microanalysis of retrieved bone tissue following maxillary sinus floor augmentation with calcium sulphateCLINICAL ORAL IMPLANTS RESEARCH, Issue 8 2008Nicola Slater Abstract Objectives: To investigate the presence and composition of residual bone graft substitute material in bone biopsies from the maxillary sinus of human subjects, following augmentation with calcium sulphate (CaS). Material and methods: Bone cores were harvested from the maxillary sinus of patients who had undergone a sinus lift procedure using CaS G170 granules 4 months after the initial surgery. Samples from seven patients, which contained residual biomaterial particles, were examined by field emission scanning electron microscopy and energy dispersive X-ray spectroscopy was used to determine the composition of the remaining bone graft substitute material. Results: Residual graft material occurred in isolated areas surrounded by bone and consisted of individual particles up to 1 mm in length and smaller spherical granules. On the basis of 187 separate point analyses, the residual material was divided into three categories (A, B and C) consisting of: A, mainly CaS (S/P atomic% ratio ,2.41); B, a heterogeneous mixture of CaS and calcium phosphate (S/P=0.11,2.4) and C, mainly calcium phosphate (S/P,0.11; C), which had a mean Ca : P ratio of 1.63±0.2, consistent with Ca-deficient hydroxyapatite. Linescans and elemental maps showed that type C material was present in areas which appeared dense and surrounded, or were adjacent to, more granular CaS-containing material, and also occurred as spherical particles. The latter could be disintegrating calcium phosphate in the final stages of the resorption process. Conclusions: CaS resorption in the human maxillary sinus is accompanied by CaP precipitation which may contribute to its biocompatibility and rapid replacement by bone. [source] Preparation of Functionalized Polysilsesquioxane and Polysilsesquioxane-Metal Nanoparticle Composite Spheres,MACROMOLECULAR RAPID COMMUNICATIONS, Issue 15 2006Young Baek Kim Abstract Summary: Network polysilsesquioxane spheres made solely of poly(vinylsilsesquioxane) (PVSQ) and poly(3-mercaptopropylsilsesquioxane) (PMPSQ) were prepared from heterogeneous mixtures of triethylamine (TEA), water, and either vinyltrimethoxysilane (VTMS), or 3-mercaptopropyltrimethoxysilane (MPTMS). The microscopic, macroscopic observations, and the relationship between the diameters of spheres and the amount of each ingredient in the reaction system, showed that spheres formed via a mechanism similar to emulsion polymerization and suspension polymerization, depending on the reaction conditions. Diameters of spheres could be controlled from tens of nanometers to a few micrometers by adjusting the amounts of TEA, water, and a surfactant. Heating aqueous solutions of metal ions with these spheres produced polysilsesquioxanes (PSQ)-metal nanoparticle composite spheres. The spheres prepared in this study were characterized by scanning electron microscopy, transmission electron microscopy, solid state NMR spectroscopy, IR spectroscopy, elemental analysis, and differential thermal analysis. These spheres would be useful in recovering metals from their ionic solutions and probes after chemical modifications. An image of the spheres of poly(vinylsilsesquioxane)-gold nanoparticle composite. [source] Effect of Nano-Aluminum and Fumed Silica Particles on Deflagration and Detonation of NitromethanePROPELLANTS, EXPLOSIVES, PYROTECHNICS, Issue 5 2009Justin Abstract The heterogeneous interaction between nitromethane (NM), particles of nanoscale aluminum (38 and 80,nm diameter), and fumed silica is examined in terms of the deflagration and detonation characteristics. Burning rates are quantified as functions of pressure using an optical pressure vessel up to 14.2,MPa, while detonation structure is characterized in terms of failure diameter. Nitromethane is gelled using fumed silica (CAB-O-SIL®), as well as by the nanoaluminum particles themselves. Use of nanoaluminum particles with fumed silica slightly increases burning rates compared to the use of larger diameter Al particles; however distinct increases in burning rates are found when CAB-O-SIL is removed and replaced with more energetic aluminum nanoparticles, whose high surface area allows them to also act as the gellant. Mixtures including fumed silica yield a reduced burning rate pressure exponent compared to neat NM, while mixtures of aluminum particles alone show a significant increase. Failure diameters of mixture detonations are found to vary significantly as a function of 38,nm aluminum particle loading, reducing more than 50% from that of neat nitromethane with 12.5% (by mass) aluminum loading. Failure diameter results indicate a relative minimum with respect to particle separation (% loading) which is not observed in other heterogeneous mixtures. [source] Different effects of cardiac versus skeletal muscle regulatory proteins on in vitro measures of actin filament speed and forceTHE JOURNAL OF PHYSIOLOGY, Issue 3 2005Emilie Warner Clemmens Mammalian cardiac and skeletal muscle express unique isoforms of the thin filament regulatory proteins, troponin (Tn) and tropomyosin (Tm), and the significance of these different isoforms in thin filament regulation has not been clearly identified. Both in vitro and skinned cellular studies investigating the mechanism of thin filament regulation in striated muscle have often used heterogeneous mixtures of Tn, Tm and myosin isoforms, and variability in reported results might be explained by different combinations of these proteins. Here we used in vitro motility and force (microneedle) assays to investigate the influence of cardiac versus skeletal Tn and Tm isoforms on actin,heavy meromyosin (HMM) mechanics. When interacting with skeletal HMM, thin filaments reconstituted with cardiac Tn/Tm or skeletal Tn/Tm exhibited similar speed,calcium relationships and significantly increased maximum speed and force per filament length (F/l) at pCa 5 (versus unregulated actin filaments). However, augmentation of F/l was greater with skeletal regulatory proteins. Reconstitution of thin filaments with the heterogeneous combination of skeletal Tn and cardiac Tm decreased sliding speeds at all [Ca2+] relative to thin filaments with skeletal Tn/Tm. Finally, for filaments reconstituted with any heterogeneous mix of Tn and Tm isoforms, force was not potentiated over that of unregulated actin filaments. Combined the results suggest (1) that cardiac regulatory proteins limit the allosteric enhancement of force, and (2) that Tn and Tm isoform homogeneity is important when studying Ca2+ regulation of crossbridge binding and kinetics as well as mechanistic differences between cardiac and skeletal muscle. [source] | ||||||||||||||||