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Different Steps (different + step)
Selected AbstractsContribution of mass spectrometry to the study of the Maillard reaction in foodMASS SPECTROMETRY REVIEWS, Issue 4 2005Laurent B. Fay Abstract The Maillard reaction or non-enzymatic browning corresponds to a set of reactions occurring between amines and carbonyl compounds, especially reducing sugars. The Maillard reaction is known to occur in heated, dried, or stored foods and in vivo in mammalian organisms. In food, the Maillard reaction is responsible for changes in colour, flavor, and nutritive value but also for the formation of stabilizing and mutagenic compounds. Because of the complexity of the Maillard reaction, mass spectrometry, coupled or not to separation techniques, is a key tool in this research area and we will review in this article the contribution of mass spectrometry to the understanding of this reaction. Different steps of Maillard reaction will be described and the importance and the role played by mass spectrometry will be highlighted. In addition, different approaches to investigate the Maillard reaction from the formation of Amadori products (early Maillard reaction product) to the flavor and melanoidin production will also be covered. © 2004 Wiley Periodicals, Inc., Mass Spec Rev 24:487,507, 2005 [source] Risk Mapping and Key Risk Indicators in Operational Risk ManagementECONOMIC NOTES, Issue 2 2005Sergio Scandizzo In this article I describe a methodology for the mapping of Operational Risk with the objective of identifying the risks inherent in the different steps of a business process, selecting a set of variables providing an estimate for the likelihood and the severity of operational risk (Key Risk Indicators , KRIs) and designing the most appropriate control activities. I then present two examples of how the methodology described can be applied to map risks and of how a set of relevant KRIs can be identified in the front office of a trading business and in the back office of a lending business. Finally, I discuss how the information conveyed by the KRIs can be organised and summarised in order to provide a comprehensive look at the risk profile of the various business lines. The structured presentation of KRIs covering the business processes of a bank is what we call an Operational Risk Scorecard. [source] Detection of C Reactive Protein (CRP) in Serum by an Electrochemical Aptamer-Based Sandwich AssayELECTROANALYSIS, Issue 11 2009Sonia Centi Abstract A disposable electrochemical assay involving magnetic particles and carbon-based screen-printed electrodes (SPCEs) was developed for the detection of C Reactive Protein (CRP). CRP is a plasma protein and is among the most expressed proteins in acute phase inflammation cases, being a known biomarker for inflammatory states. The assay was based on a sandwich format in which a RNA aptamer was coupled to a monoclonal antibody and alkaline phosphatase (AP) was used as enzymatic label. After the sandwich assay, the modified magnetic beads were captured by a magnet on the surface of a graphite working electrode and the electrochemical detection was thus achieved through the addition of the AP substrate (,-naphthyl-phosphate) and ,-naphthol produced during the enzymatic reaction was detected using differential pulse voltammetry (DPV). The parameters influencing the different steps of the assay were optimized in order to reach the best sensitivity and specificity. With the optimized conditions, the assay was applied to the analysis of CRP free serum and serum samples. [source] Comparison of the effectiveness of five extraction methods for Clavibacter michiganensis subsp. sepedonicus and Ralstonia solanacearum from potato tubersEPPO BULLETIN, Issue 2 2001J. Martin In the EU Control Directives, the recommended extraction procedure for testing potatoes for Clavibacter michiganensis subsp. sepedonicus and Ralstonia solanacearum comprises incubation followed by differential centrifugation. This method can be qualified as complex because of the number of different steps required. This study evaluates five different extraction methods for each bacterium from both a technical point of view and for the quality of the results. Results showed that in the case of C. m. sepedonicus the clarification step should be avoided. The incubation/shaking method with three subsamples gives at least as satisfactory results as the official EU procedure. It also has other advantages, facilitating immunofluorescence readings due to the reduced quantity of plant debris, and improving the speed and the reliability of the analysis. [source] POSTMATING SEXUAL SELECTION: ALLOPATRIC EVOLUTION OF SPERM COMPETITION MECHANISMS AND GENITAL MORPHOLOGY IN CALOPTERYGID DAMSELFLIES (INSECTA: ODONATA)EVOLUTION, Issue 2 2004A. Cordero Rivera Abstract Postmating sexual selection theory predicts that in allopatry reproductive traits diverge rapidly and that the resulting differentiation in these traits may lead to restrictions to gene flow between populations and, eventually, reproductive isolation. In this paper we explore the potential for this premise in a group of damselflies of the family Calopterygidae, in which postmating sexual mechanisms are especially well understood. Particularly, we tested if in allopatric populations the sperm competition mechanisms and genitalic traits involved in these mechanisms have indeed diverged as sexual selection theory predicts. We did so in two different steps. First, we compared the sperm competition mechanisms of two allopatric populations of Calopteryx haemorrhoidalis (one Italian population studied here and one Spanish population previously studied). Our results indicate that in both populations males are able to displace spermathecal sperm, but the mechanism used for sperm removal between both populations is strikingly different. In the Spanish population males seem to empty the spermathecae by stimulating females, whereas in the Italian population males physically remove sperm from the spermathecae. Both populations also exhibit differences in genital morphometry that explain the use of different mechanisms: the male lateral processes are narrower than the spermathecal ducts in the Italian population, which is the reverse in the Spanish population. The estimated degree of phenotypic differentiation between these populations based on the genitalic traits involved in sperm removal was much greater than the differentiation based on a set of other seven morphological variables, suggesting that strong directional postmating sexual selection is indeed the main evolutionary force behind the reproductive differentiation between the studied populations. In a second step, we examined if a similar pattern in genital morphometry emerge in allopatric populations of this and other three species of the same family (Calopteryx splendens, C. virgo and Hetaerina cruentata). Our results suggest that there is geographic variation in the sperm competition mechanisms in all four studied species. Furthermore, genitalic morphology was significantly divergent between populations within species even when different populations were using the same copulatory mechanism. These results can be explained by probable local coadaptation processes that have given rise to an ability or inability to reach and displace spermathecal sperm in different populations. This set of results provides the first direct evidence of intraspecific evolution of genitalic traits shaped by postmating sexual selection. [source] Epidermal Growth Factor Regulates Amino Acid Transport in Chick Embryo Hepatocytes via Protein Kinase CEXPERIMENTAL PHYSIOLOGY, Issue 4 2000Maria Marino System A-mediated amino acid transport, activation of different steps of signal transduction and involvement of different isoforms of protein kinase C (PKC) have been investigated in chick embryo hepatocytes after epidermal growth factor (EGF) stimulation. EGF rapidly (10 min) increased the rate of aminoisobutyric acid (AIB) uptake in chick embryo hepatocytes freshly isolated on the 19th day of embryonic life, while no change was detectable at other embryonal stages. The growth factor stimulation was abolished by PKC and tyrosine kinase inhibitors and was mimicked by 4-phorbol-12-myristate-13-acetate, dimethyl-2 (PMA). EGF treatment did not modify the phosphorylation of the , isoform of phospholipase C (PLC-,), and inositol trisphosphate (IP3) and intracellular calcium levels, but it induced an increase in PKC activity. Our data show that EGF regulates amino acid uptake, via PKC and without PLC-, activation, only in the last period of chick embryo hepatocyte development. The effects of growth factor on PKC activity suggest the involvement of PKC-, and -, isoforms in EGF modulation of amino acid transport. [source] Functional transitions of F0F1 -ATPase mediated by the inhibitory peptide IF1 in yeast coupled submitochondrial particlesFEBS JOURNAL, Issue 10 2004Mikhail Galkin The mechanism of inhibition of yeast F0F1 -ATPase by its naturally occurring protein inhibitor (IF1) was investigated in submitochondrial particles by studying the IF1-mediated ATPase inhibition in the presence and absence of a protonmotive force. In the presence of protonmotive force, IF1 added during net NTP hydrolysis almost completely inhibited NTPase activity. At moderate IF1 concentration, subsequent uncoupler addition unexpectedly caused a burst of NTP hydrolysis. We propose that the protonmotive force induces the conversion of IF1-inhibited F0F1 -ATPase into a new form having a lower affinity for IF1. This form remains inactive for ATP hydrolysis after IF1 release. Uncoupling simultaneously releases ATP hydrolysis and converts the latent form of IF1-free F0F1 -ATPase back to the active form. The relationship between the different steps of the catalytic cycle, the mechanism of inhibition by IF1 and the interconversion process is discussed. [source] French adaptation and preliminary validation of a questionnaire to evaluate understanding of informed consent documents in phase I biomedical researchFUNDAMENTAL & CLINICAL PHARMACOLOGY, Issue 1 2006Adeline Paris Abstract The content of informed consent documents (ICD) is a crucial element in the process of providing information to participants in biomedical research. Clear comprehension of the information, i.e. the ability to understand its meaning and its consequences, is of utmost importance. The objective of this study was to describe the different steps in the French adaptation and preliminary validation of the Qualité de Compréhension des Formulaires d'information et de consentement (QCFic) questionnaire (http://www.lyon.inserm.fr/cic-grenoble) based on the American Quality of Informed Consent (QuIC) questionnaire. Adaptation and preliminary validation of the QuIC for use in France was composed of five principal steps: translation, scientific validation, lexical validation, edition of gold-standard answers and a pilot study. Each stage was conducted by independent groups of experts, under the coordination of the study board. Thirteen questions were added and one was suppressed. Two steps were required for the scientific validation and for lexical validation, 21 modifications were proposed. Relative to gold-standard answers, the three experts gave the same answer for 24 questions and for nine other questions, two of the three gave identical answers, which were validated by the study board. Results of a pilot study showed a global QCFic score of 88.99 (84.13,90.92) and no specific commentary was made about the content of the questions, so no more modification needed to be made. A preliminary validated French questionnaire, the QCFic, is now available to evaluate the quality of an informed consent document in phase I clinical trials. It is quick and easy to use. [source] Radial migration of developing microglial cells in quail retina: A confocal microscopy studyGLIA, Issue 3 2004Ana Sánchez-López Abstract Microglial cells spread within the nervous system by tangential and radial migration. The cellular mechanism of tangential migration of microglia has been described in the quail retina but the mechanism of their radial migration has not been studied. In this work, we clarify some aspects of this mechanism by analyzing morphological features of microglial cells at different steps of their radial migration in the quail retina. Microglial cells migrate in the vitreal half of the retina by successive jumps from the vitreal border to progressively more scleral levels located at the vitreal border, intermediate regions, and scleral border of the inner plexiform layer (IPL). The cellular mechanism used for each jump consists of the emission of a leading thin radial process that ramifies at a more scleral level before retraction of the rear of the cell. Hence, radial migration and ramification of microglial cells are simultaneous events. Once at the scleral border of the IPL, microglial cells migrate through the inner nuclear layer to the outer plexiform layer by another mechanism: they retract cell processes, become round, and squeeze through neuronal bodies. Microglial cells use radial processes of s-laminin-expressing Müller cells as substratum for radial migration. Levels where microglial cells stop and ramify at each jump are always interfaces between retinal strata with strong tenascin immunostaining and strata showing weak or no tenascin immunoreactivity. When microglial cell radial migration ends, tenascin immunostaining is no longer present in the retina. These findings suggest that tenascin plays a role in the stopping and ramification of radially migrating microglial cells. © 2004 Wiley-Liss, Inc. [source] Determinants of vertebrate invasion success in Europe and North AmericaGLOBAL CHANGE BIOLOGY, Issue 9 2006JONATHAN M. JESCHKE Abstract Species that are frequently introduced to an exotic range have a high potential of becoming invasive. Besides propagule pressure, however, no other generally strong determinant of invasion success is known. Although evidence has accumulated that human affiliates (domesticates, pets, human commensals) also have high invasion success, existing studies do not distinguish whether this success can be completely explained by or is partly independent of propagule pressure. Here, we analyze both factors independently, propagule pressure and human affiliation. We also consider a third factor directly related to humans, hunting, and 17 traits on each species' population size and extent, diet, body size, and life history. Our dataset includes all 2362 freshwater fish, mammals, and birds native to Europe or North America. In contrast to most previous studies, we look at the complete invasion process consisting of (1) introduction, (2) establishment, and (3) spread. In this way, we not only consider which of the introduced species became invasive but also which species were introduced. Of the 20 factors tested, propagule pressure and human affiliation were the two strongest determinants of invasion success across all taxa and steps. This was true for multivariate analyses that account for intercorrelations among variables as well as univariate analyses, suggesting that human affiliation influenced invasion success independently of propagule pressure. Some factors affected the different steps of the invasion process antagonistically. For example, game species were much more likely to be introduced to an exotic continent than nonhunted species but tended to be less likely to establish themselves and spread. Such antagonistic effects show the importance of considering the complete invasion process. [source] Tissue Engineering Strategies Designed to Realize the Endogenous Regenerative Potential of Peripheral NervesADVANCED MATERIALS, Issue 46 2009Vivek Mukhatyar Abstract http://doi.wiley.com/10.1002/adma.v21:32/33 Bridging peripheral nerve gaps without the use of autografts has significant clinical importance. But in order to rationally design novel scaffolds, a good understanding of the nerve regeneration process is vital. Appropriate amount of structural and chemical cues are required to stimulate the endogenous mechanisms of repair and functional recovery. Synthetic and natural materials present various opportunities to induce the growth of supporting cells as well as promote axon regeneration. An overview of tissue engineering strategies currently being explored that stimulate the different steps of the regenerative sequence is presented. [source] An eddy current integral formulation on parallel computer systemsINTERNATIONAL JOURNAL FOR NUMERICAL METHODS IN ENGINEERING, Issue 9 2005Raffaele Fresa Abstract In this paper, we show how an eddy current volume integral formulation can be used to analyse complex 3D conducting structures, achieving a substantial benefit from the use of a parallel computer system. To this purpose, the different steps of the numerical algorithms in view of their parallelization are outlined to enlighten the merits and the limitations of the proposed approach. Numerical examples are developed in a parallel environment to show the effectiveness of the method. Copyright © 2004 John Wiley & Sons, Ltd. [source] Oxidation of diclofenac sodium by diperiodatoargantate(III) in aqueous alkaline medium and its determination in urine and blood by kinetic methodsINTERNATIONAL JOURNAL OF CHEMICAL KINETICS, Issue 6 2010P. N. Naik The kinetics and oxidation of diclofenac sodium (DFS) by diperiodatoargentate(III) (DPA) in alkaline medium at 298 K and at a constant ionic strength of 0.60 mol dm,3 were studied spectrophotometrically. The oxidation products were [2-(2,6-dicloro-phynylamino)-phenyl]-methenol and Ag(I), identified by LC-ESI-MS and IR spectral studies. The reaction between DFS and DPA in alkaline medium exhibits 1:1 stoichiometry. The reaction is first order in [DPA] and has a less than unit order dependence each in [DFS] and [alkali]. Increasing concentrations of IO,4 retard the reaction. The active species of DPA proposed to be monoperiodatoargentate(III), and a mechanism is suggested. The rate constants involved in the different steps of the mechanism were determined and are discussed. The activation parameters with respect to a rate-limiting step of the mechanism were determined. The thermodynamic quantities were also determined. Using the oxidation of DFS by DPA, DFS was analyzed by kinetic methods in urine and blood sample. The proposed method enables DFS analysis in the range from 5.0 × 10,5 to 5.0 × 10,3 mol dm,3. © 2010 Wiley Periodicals, Inc. Int J Chem Kinet 42: 336,346, 2010 [source] Life cycle assessment of a PPV plant applied to an existing SUW management systemINTERNATIONAL JOURNAL OF ENERGY RESEARCH, Issue 5 2003Francesco Di Maria Abstract The huge amount of wastes produced by modern and developed countries involves important aspects of economical, social and technical fields and also of the environment. For this reason, different technologies have been proposed for trying to reduce the impact of waste management and disposal. Generally waste management system consists of different steps like selective collection, recycling and reuse operation, energy recovery from waste and landfilling. A new technology proposed for thermal waste treatment is the plasma pyrolysis vetrification (PPV). This system seems to have interesting perspective due to the possibility of thermal treatment of dangerous slag or waste producing inactivate vetrified substances that can be landfilled or used as building materials with no impact on the environment. In this study, the effect of the application of a PPV plant on an existing waste management system was evaluated with a life cycle assessment (LCA) analysis. All the activities connected to the existing system have been carefully analysed by collecting a large quantity of experimental data. Some assumptions have been made, in particular, on the PPV plant performance. LCA analysis results illustrate how the environmental benefits arising from the adoption of the new technology, concerns only few aspects of the whole system. Copyright © 2003 John Wiley & Sons, Ltd. [source] Theoretical study of hydrogen-bonded complexes of benzene with hydrides of astrochemical interestINTERNATIONAL JOURNAL OF QUANTUM CHEMISTRY, Issue 3 2008M. Nait Achour Abstract Post Hartree,Fock and DFT calculations have been performed for studying the possibility for a benzene support to be linked to various hydrides through a quasi Bz···HA bond. Interaction energy of compounds, including CH bonds (CH4, CH3F, CH2O, CHN, CHNO), NH bonds (NH3, NH2F, NHC, NHCO, NH3O), and OH bonds (OH2, OHF, NCOH), were evaluated, taking basis set superposition error (BSSE) and zero point vibrational energy (ZPVE) corrections into account. Numerical convergence of results with respect to the ingredients included at different steps of theory (basis set, DFT functionals, correlation treatments, geometry optimization) was tested mainly on the example of the water adduct and, for comparison, the Bz···H3O+ system containing a cation instead of a neutral molecule. A rather large range of adsorption energies is obtained, from about 1 kcal/mol for methane to more than 6 kcal/mol for cyanic acid, according to the acidic character of the adsorbed species in each family of Bz···HA bonds. Some consequences for astrophysical problems involving PAHs in the interstellar medium are pointed out. © 2007 Wiley Periodicals, Inc. Int J Quantum Chem, 2008 [source] Diverse Asymmetric Quinolizidine Synthesis: A Stereodivergent One-Pot ApproachADVANCED SYNTHESIS & CATALYSIS (PREVIOUSLY: JOURNAL FUER PRAKTISCHE CHEMIE), Issue 2-3 2010Wei Zhang Abstract A diverse stereodivergent organocatalytic one-pot addition/cyclization/annulation sequence to optically active quinolizidine derivatives from easily available starting materials is presented. The one-pot sequence relies on a pyrrolidine-catalyzed enantioselective conjugate addition of electron-deficient amide ,-carbons to ,,,-unsaturated aldehydes, spontaneous hemiaminal formation and acid-catalyzed/mediated N -acyliminium ion cyclization to give the quinolizidine framework. Simple tuning of the reaction conditions in the N -acyliminuim ion cyclization step provides a diastereomeric switch, which gives access to both of the two bridgehead epimers through kinetic, thermodynamic or chelation control. The methodology display a broad substrate scope that is demonstrated by the stereoselective formation of indolo-, thieno-, benzofuro-, furo- and different benzoquinolizidine derivatives with high atom efficiency, up to >99% ee and up to >95:5 dr. Due to its efficiency, synthetic diversity and operational simplicity, this protocol has the potential to find important use as a key step in natural product synthesis, biochemistry and pharmaceutical science. The stereochemical outcome of the one-pot sequence was investigated, and the mechanism and origin of stereoselectivity of the different steps is discussed. [source] Sonographic assessment of clubfootJOURNAL OF CLINICAL ULTRASOUND, Issue 5 2004Cosimo Gigante MD Abstract Purpose This study was performed to develop a standardized methodology for the sonographic assessment of clubfoot at birth and at the end of both conservative treatment and surgical correction. Methods Forty-two congenital clubfeet and 42 normal feet were examined sonographically in the position of spontaneous alignment and during passive manual correction. Scans along 4 planes provided information relevant to the assessment: sagittal posterior, sagittal anterior, coronal lateral, and transverse. Results Sagittal posterior sonograms demonstrated the progressive gain of dorsiflexion ability during the different steps of treatment for clubfoot. Sagittal anterior sonograms could not demonstrate the normal alignment of the navicular in clubfeet because of the bone's medial displacement. On transverse sonograms, the talar head and the medially displaced navicular may lie on the same plane, depending on the severity of the deformity. Coronal lateral sonograms provided for estimation of the relationships between the calcaneus and cuboid, which were described by the calcaneal-cuboid angle. Conclusions Sonography is a promising technique for assessment and monitoring of clubfoot during treatment. The method described here yields accurate and reproducible information about the anatomy of the nonossified clubfoot, helping the orthopedic team decide on appropriate treatment steps. © 2004 Wiley Periodicals, Inc. J Clin Ultrasound 32:235,242, 2004; Published online in Wiley InterScience (www.interscience.wiley.com). DOI: 10.1002/jcu.20022 [source] Modeling aspects of mechanisms for reactions catalyzed by metalloenzymesJOURNAL OF COMPUTATIONAL CHEMISTRY, Issue 14 2001P. E. M. Siegbahn Different models to treat metal-catalyzed enzyme reactions are investigated. The test case chosen is the recently suggested full catalytic cycle of manganese catalase including eight different steps. This cycle contains OO and OH activations, as well as electron transfer steps and redox active reactions, and is therefore believed to be representative of many similar systems. Questions concerning modeling of ligands and the accuracy of the computational model are studied. Imidazole modeling of histidines are compared to ammonia modeling, and formate modeling compared to acetate modeling of glutamates. The basis set size required for the geometry optimization and for the final energy evaluation is also investigated. The adequacy of the model is judged in relation to the inherent accuracy achievable with the hybrid DFT method B3LYP. © 2001 John Wiley & Sons, Inc. J Comput Chem 22: 1634,1645, 2001 [source] ,-tocopherol, an exogenous factor of adult hippocampal neurogenesis regulationJOURNAL OF NEUROSCIENCE RESEARCH, Issue 4 2003Tiziana Cecchini Abstract In previous work, we found that adult hippocampal neurogenesis in rat is affected by vitamin E deficiency. Because vitamin E deficiency is a complex condition involving numerous biological systems, it is possible that its effect on postnatal new neuron production could be mediated by unknown changes in different factors that in turn play a role in this process. To clarify if vitamin E plays a direct role in regulating hippocampal neurogenesis, we studied the neurogenesis in adult control rats and in adult rats under supplementation with ,-tocopherol, the most important compound of vitamin E. The ,-tocopherol level in control and supplemented rats was monitored. Qualitative and quantitative analysis of cell proliferation and death was carried out and expression of immature neuron markers PSA-NCAM, TUC 4, and DCX was investigated in hippocampus dentate gyrus. ,-Tocopherol levels increased significantly in both plasma and brain after supplementation. Cell proliferation was inhibited in ,-tocopherol-supplemented rats, the number of dying cells was reduced, and the number of cells expressing the immature neuron markers was increased. The results obtained confirm and extend the idea that vitamin E is an exogenous factor playing a direct role in regulation of different steps of adult hippocampal neurogenesis. Some hypotheses about the possible mechanisms underlying the complex action of ,-tocopherol, related to its antioxidant and molecule-specific non-antioxidant properties, are proposed and discussed. © 2003 Wiley-Liss, Inc. [source] Autologous nucleus pulposus primes T cells to develop into interleukin-4-producing effector cells: An experimental study on the autoimmune properties of nucleus pulposusJOURNAL OF ORTHOPAEDIC RESEARCH, Issue 1 2009Andrea Geiss Abstract An autoimmune response to herniated nucleus pulposus has been proposed to constitute a pathophysiologic mechanism for inducing sciatica based on the fact that nucleus pulposus under normal conditions is excluded from the development of immunological tolerance. The manifestation of an autoimmune response comprises different steps starting with antigen capture, continuing with activation of T helper (TH) cells and ending with production of autoantibodies. Activated TH cells differentiate into either TH1 cells, predominately producing proinflammatory cytokines such as interferon , (IFN,) or a TH2 subset mainly producing anti-inflammatory cytokines such as interleukin-4 (IL-4). The aim of the present study was to examine if exposure of autologous nucleus pulposus (NP) to the immune system for 3 weeks is potent enough to prime TH cells to differentiate into TH2 cells. The study was performed in a pig model allowing the exposure of NP to the immune system. To assess the polarization of TH cells the intracellular production of IFN, and IL-4 was measured in T cells by using flow cytometry. The revealed predominant production of IL-4 together with low production of IFN, in T cells after NP exposure to the immune system indicates that nucleus pulposus may prime TH cells to develop into IL-4-producing TH2 cells after being exposed to the immune system, for example, in association with disc herniation. © 2008 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 27:97,103, 2009 [source] Poly(styrene- co -glycerol dimethacrylate): Synthesis, characterization, and application as a resin for gel-phase peptide synthesisJOURNAL OF POLYMER SCIENCE (IN TWO SECTIONS), Issue 19 2005Michael Roice Abstract An efficient cross-linked polymer support for solid-phase synthesis was prepared by introducing glycerol dimethacrylate cross-linker to polystyrene network using free radical aqueous suspension polymerization. The support was characterized by various spectroscopic methods. Morphological feature of the resin was analyzed by microscopy. The polymerization reaction was investigated with respect to the effect of amount of cross-linking agent, which in turn vary the swelling, loading, and the mechanical stability of the resin. The solvent uptake of the polymer was studied in relation to cross-linking and compared with Merrifield resin. The stability of the resin was tested in different synthetic conditions used for solid-phase peptide synthesis. Hydroxy group of the support was derivatized to chloro and then amino groups using different reagents and reaction conditions. Efficiency of the support was tested and compared with TentaGelÔ resin by following different steps involved in the synthesis of the 65,74 fragment of acyl carrier protein. The results showed that the poly(styrene- co -glycerol dimethacrylate) (GDMA-PS) is equally efficient as TentaGel resin in peptide synthesis. The purity of the peptides was analyzed by HPLC and identities were determined by mass spectroscopy and amino acid analysis. © 2005 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 43: 4382,4392, 2005 [source] Sulfur fertilization and light exposure during storage are critical determinants of the nutritional value of ready-to-eat friariello campano (Brassica rapa L. subsp. sylvestris)JOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 13 2009Giancarlo Barbieri Abstract BACKGROUND: The nutritional value of fresh vegetables can be affected at different steps within the field-to-market pipeline. Both pre- and post-harvest factors should be considered in order to increase the produce quality and to preserve it until final consumption. In this study the effects of sulfur nutrition during plant growth and light exposure during storage on the nutritional value of ready-to-eat friariello campano (Brassica rapa L. subsp. sylvestris L. Janch. var. esculenta Hort.) were studied. RESULTS: Fresh weight loss was higher in light-storage treatment. During storage, light exposure reduced leaf nitrate content, although no effect could be attributed to sulfur nutrition. Sulfur increased both lipophilic and hydrophilic antioxidant activity. The hydrophilic antioxidant activity linearly decreased during storage, whereas the lipophilic activity increased over time. However, no differences could be attributed to light exposure during storage for this parameter. Results on colorimetric indexes and their relation with the chlorophyll content are also reported. CONCLUSIONS: Ready-to-eat friariello quality may be improved with an enhanced antioxidant activity and reduced nitrate content by combining, respectively, increased sulfur availability during plant growth and exposure to light during storage. On the other hand, light exposure caused a more rapid decline in produce fresh weight during storage. Copyright © 2009 Society of Chemical Industry [source] Chemical characterization of tomato pomaceJOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 8 2006Marcos Del Valle Abstract Tomato-processing by-product, also known as tomato pomace, consists of peel and seeds and represents around 4% of the fruit weight. If these wastes remain unused, they not only add to the disposal problem but also aggravate environmental pollution. The aim of this paper is to evaluate the chemical composition of tomato pomace collected at different steps during industrial processing in order to assess the quality of this by-product. Twenty-one tomato pomace samples were considered for analysis. Samples corresponded to different steps during tomato processing for paste: seven after pulper, five after finisher, six before turbopress and three after turbopress. Samples were analyzed for moisture content, total and soluble sugars, protein, fat, soluble and total fiber, as well as mineral content. From the results obtained we can conclude that tomato pomace composition (in dry weight basis) is as follows: 59.03% fiber, 25.73% total sugars, 19.27% protein, 7.55% pectins, 5.85% total fat and 3.92% minerals. Copyright © 2006 Society of Chemical Industry [source] Resveratrol: Preventing properties against vascular alterations and ageingMOLECULAR NUTRITION & FOOD RESEARCH (FORMERLY NAHRUNG/FOOD), Issue 5 2005Dominique Delmas Abstract Cardiovascular diseases are the leading cause of death in developed countries where the common pathological substrate underlying this process is atherosclerosis. Several new concepts have emerged in relation to mechanisms that contribute to the regulation of the vascular diseases and associated inflammatory effects. Recently, potential antioxidants (vitamin E, polyphenols) have received much attention as potential anti-atherosclerotic agents. Among the polyphenols with health benefic properties, resveratrol, a phytoalexin of grape, seem to be a good candidate protecting the vascular walls from oxidation, inflammation, platelet aggregation, and thrombus formation. In this review, we focus on the mechanism of resveratrol cardiovascular benefic effects. We analyze, in relation with the different steps of atherosclerotic process, the resveratrol properties at multiple levels, such as cellular signaling, enzymatic pathways, apoptosis, and gene expression. We show and discuss the relationship with reactive oxygen species, regulation of pro-inflammatory genes including cycloxygenases and cytokines in molecular inflammatory and aging processes, and how the regulation of these activites by resveratrol can lead to a prevention of vascular diseases. [source] Mutational Analysis of the Modulation of Tyrosinase by Tyrosinase-Related Proteins 1 and 2 In VitroPIGMENT CELL & MELANOMA RESEARCH, Issue 5 2000PRASHIELA MANGA The albino (tyrosinase, Tyrc), brown (tyrosinase-related protein 1, Tyrp1b) and slaty (tyrosinase-related protein 2, tyrp2slt) loci are all involved in the regulation of melanogenesis. Phenotypes of inbred mice mutant at two or more of these loci are not always explicable by simple summation of the established or suspected catalytic functions of the gene products. These phenotypes suggest that relationships among the proteins extend beyond the obvious fact that they catalyze different steps in the same melanogenic pathway, and that they may also interact intimately in such a way that a mutation in one impacts the function of the other(s). Previous studies have attributed catalytic activities to each member of this trio; however, it has been difficult to study the proteins individually, either in vivo or in tissues or cells. Therefore, we undertook to transfect the genes, in revealing combinations, into COS-7 cells (which have no melanogenic apparatus of their own) to clarify the interacting functions of their encoded proteins. Specifically, we attempted to evaluate the effects of Tyrp1 and Tyrp2 proteins on tyrosinase protein. We report evidence that Tyrp1 stabilizes tyrosinase, confirming previous observations, and, in addition, demonstrate that Tyrp1 decreases tyrosinase activity. By contrast, Tyrp2 increases tyrosinase activity by stabilizing the protein. We conclude that both Tyrp1 and Tyrp2, in addition to other catalytic functions they may possess, act together to modulate tyrosinase activity. [source] Relative contributions of nine genes in the pathway of histidine biosynthesis to the control of free histidine concentrations in Arabidopsis thalianaPLANT BIOTECHNOLOGY JOURNAL, Issue 6 2009Jonathan D. Rees Summary Despite the functional importance of histidine (His) as an essential amino acid in proteins and as a metal-coordinating ligand, comparatively little is known about the regulation of its biosynthesis in plants and the potential for metabolic engineering of this pathway. To investigate the contribution of different steps in the pathway to overall control of His biosynthesis, nine His biosynthetic genes were individually over-expressed in Arabidopsis thaliana to determine their effects on free amino acid pools. Constitutive, CaMV 35S -driven over-expression of the cDNAs encoding either isoform of ATP-phosphoribosyltransferase (ATP-PRT), the first enzyme in the pathway, was sufficient to increase the pool of free His by up to 42-fold in shoot tissue of Arabidopsis, with negligible effect on any other amino acid. In contrast, over-expression of cDNAs for seven other enzymes in the biosynthetic pathway had no effect on His content, suggesting that control of the pool of free His resides largely with ATP-PRT activity. Over-expression of ATP-PRT and increased His content had a negative pleiotropic effect on plant biomass production in 35S:PRT1 lines, but this effect was not observed in 35S:PRT2 lines. In the presence of 100 µM Ni, which was inhibitory to wild-type plants, a strong positive correlation was observed between free His content and biomass production, indicating that the metabolic cost of His overproduction was outweighed by the benefit of increased tolerance to Ni. His-overproducing plants also displayed somewhat elevated tolerance to Co and Zn, but not to Cd or Cu, indicating chemical selectivity in intracellular metal binding by His. [source] A unified mechanism for protein folding: Predetermined pathways with optional errorsPROTEIN SCIENCE, Issue 3 2007Mallela M.G. Krishna Abstract There is a fundamental conflict between two different views of how proteins fold. Kinetic experiments and theoretical calculations are often interpreted in terms of different population fractions folding through different intermediates in independent unrelated pathways (IUP model). However, detailed structural information indicates that all of the protein population folds through a sequence of intermediates predetermined by the foldon substructure of the target protein and a sequential stabilization principle. These contrary views can be resolved by a predetermined pathway,optional error (PPOE) hypothesis. The hypothesis is that any pathway intermediate can incorporate a chance misfolding error that blocks folding and must be reversed for productive folding to continue. Different fractions of the protein population will then block at different steps, populate different intermediates, and fold at different rates, giving the appearance of multiple unrelated pathways. A test of the hypothesis matches the two models against extensive kinetic folding results for hen lysozyme which have been widely cited in support of independent parallel pathways. The PPOE model succeeds with fewer fitting constants. The fitted PPOE reaction scheme leads to known folding behavior, whereas the IUP properties are contradicted by experiment. The appearance of a conflict with multipath theoretical models seems to be due to their different focus, namely on multitrack microscopic behavior versus cooperative macroscopic behavior. The integration of three well-documented principles in the PPOE model (cooperative foldons, sequential stabilization, optional errors) provides a unifying explanation for how proteins fold and why they fold in that way. [source] The oligomeric state and stability of the mannitol transporter, EnzymeIImtl, from Escherichia coli: A fluorescence correlation spectroscopy studyPROTEIN SCIENCE, Issue 8 2006Gertjan Veldhuis Abstract Numerous membrane proteins function as oligomers both at the structural and functional levels. The mannitol transporter from Escherichia coli, EnzymeIImtl, is a member of the phosphoenolpyruvate-dependent phosphotransferase system. During the transport cycle, mannitol is phosphorylated and released into the cytoplasm as mannitol-1-phosphate. Several studies have shown that EIImtl functions as an oligomeric species. However, the oligomerization number and stability of the oligomeric complex during different steps of the catalytic cycle, e.g., substrate binding and/or phosphorylation of the carrier, is still under discussion. In this paper, we have addressed the oligomeric state and stability of EIImtl using fluorescence correlation spectroscopy. A functional double-cysteine mutant was site-specifically labeled with either Alexa Fluor 488 or Alexa Fluor 633. The subunit exchange of these two batches of proteins was followed in time during different steps of the catalytic cycle. The most important conclusions are that (1) in a detergent-solubilized state, EIImtl is functional as a very stable dimer; (2) the stability of the complex can be manipulated by changing the intermicellar attractive forces between PEG-based detergent micelles; (3) substrate binding destabilizes the complex whereas phosphorylation increases the stability; and (4) substrate binding to the phosphorylated species partly antagonizes the stabilizing effect. [source] OC1 Effects of Cryopreservation on the Expression of Glut-3, Glut-5 and As-A Proteins in Iberian Boar Sperm MembranesREPRODUCTION IN DOMESTIC ANIMALS, Issue 2006S Sancho In order to determine the injure produced in boar spermatozoa through cryopreservation process, we analyzed the expression of the hexose transporters Glut-3 and Glut-5 and the zona pellucida binding protein As-A (P68) in three different steps of the freezing-thawed protocol: at 17°C (fresh BTS-diluted semen, 1 : 2 v/v, step 1), at 5°C (after glycerol addition; step 2), and post-thawing (step 3). All sperm analyses were carried out with immunogold techniques under electronic microscopy. For this study eight healthy post-pubertal Iberian boars were submitted to a collection of twice per week through 3 months, evaluating two ejaculates from each boar. Glut-3 maintains the expression in the acrosome region post-thawing but not along the tail where is reduced. The expression of Glut-5 and As-A is majority located at the post-acrosome region of the spermatozoa at step 1, but in step 2 and step 3 this expression is relocated to sperm tail area. In conclusion, while cryopreservation affects the localization and the expression of Glut-3 and Glut-5, its fertilizing capacity is not significantly reduced. The stabilization of boar semen at 5°C was found to be the most crucial step for sperm survival. [source] The role of inflammatory and parenchymal cells in acute pancreatitis,THE JOURNAL OF PATHOLOGY, Issue 3 2007A Vonlaufen Abstract The infiltration of inflammatory cells into the pancreas is an early and central event in acute pancreatitis that promotes local injury and systemic complications of the disease. Recent research has yielded the important finding that resident cells of the pancreas (particularly acinar and pancreatic stellate cells) play a dynamic role in leukocyte attraction via secretion of chemokines and cytokines and expression of adhesion molecules. Significant progress has been made in recent years in our understanding of the role of leukocyte movement (adhesion to the blood vessel wall, transmigration through the blood vessel wall and infiltration into the parenchyma) in the pathophysiology of acute pancreatitis. This review discusses recent studies and describes the current state of knowledge in the field. It is clear that detailed elucidation of the numerous processes in the inflammatory cascade is an essential step towards the development of improved therapeutic strategies in acute pancreatitis. Studies to date suggest that combination therapy targeting different steps of the inflammatory cascade may be the treatment of choice for this disease. Copyright © 2007 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd. [source] | |||||||||||||||||||||||||||||||