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Selected AbstractsThe role of seeds and airborne inoculum in the initiation of leaf blotch (Rhynchosporium secalis) epidemics in winter barleyPLANT PATHOLOGY, Issue 2 2010J. M. Fountaine Both airborne spores of Rhynchosporium secalis and seed infection have been implied as major sources of primary inoculum for barley leaf blotch (scald) epidemics in fields without previous history of barley cropping. However, little is known about their relative importance in the onset of disease. Results from both quantitative real-time PCR and visual assessments indicated that seed infection was the main source of inoculum in the field trial conducted in this study. Glasshouse studies established that the pathogen can be transmitted from infected seeds into roots, shoots and leaves without causing symptoms. Plants in the field trial remained symptomless for approximately four months before symptoms were observed in the crop. Covering the crop during part of the growing season was shown to prevent pathogen growth, despite the use of infected seed, indicating that changes in the physiological condition of the plant and/or environmental conditions may trigger disease development. However, once the disease appeared in the field it quickly became uniform throughout the cropping area. Only small amounts of R. secalis DNA were measured in 24 h spore-trap tape samples using PCR. Inoculum levels equivalent to spore concentrations between 30 and 60 spores per m3 of air were only detected on three occasions during the growing season. The temporal pattern and level of detection of R. secalis DNA in spore tape samples indicated that airborne inoculum was limited and most likely represented rain-splashed conidia rather than putative ascospores. [source] Effect of an epoxy octasilsesquioxane on the thermodegradation of an epoxy/amine systemPOLYMER INTERNATIONAL, Issue 1 2010Belén Montero Abstract Polyhedral oligomeric silsesquioxanes (POSS®) can be added to thermoplastic and thermostable polymers to obtain hybrid materials with only a minor tendency to suffer ignition. The aim of the work reported was to analyse the influence of an octafunctional POSS® in the pyrolysis of an epoxy/amine system as well as during the combustion process. Thermal degradation of the modified materials, with respect to the unmodified ones, was analysed using thermogravimetric analysis. As the content of POSS® increased the stability improved and the char/ceramic yields were higher. The Kissinger,Akahira,Sunose method was applied to the modified blends and it showed a decrease in the activation energy with POSS® content. Empirical kinetic models, as well as generalized master plots, were applied to explain the degradation mechanism for ternary blends. The limiting oxygen index parameter was measured to analyse the fire retardancy effect of POSS®: it increased from 24.3 to 25.4% with the addition of 2.5 wt% of POSS®. The mechanism of thermal degradation of the hybrid materials based on an epoxy resin is affected by the presence of the octaepoxy POSS®. Only small amounts of POSS® are necessary to enhance the combustion resistance of the system. Copyright © 2009 Society of Chemical Industry [source] Electrophoretically mediated microanalysis with partial filling technique and indirect or direct detection as a tool for inhibition studies of enzymatic reactionELECTROPHORESIS, Issue 7-8 2004Magdaléna Telnarová Abstract The inhibition of the model enzyme, haloalkane dehalogenase from Sphingomonas paucimobilis, was investigated by a combination of electrophoretically mediated microanalysis with a partial filling technique, followed by indirect or direct detection. In this setup, part of the capillary is filled with a buffer suitable for the enzymatic reaction (20 mM glycine buffer, pH 8.6) whereas the rest of the capillary is filled with the background electrolyte optimal for separation of substrates and products. Two different background electrolytes and corresponding detection approaches were used to show the versatility of the developed method. The inhibition effect of 1,2-dichloroethane on the dehalogenation of brominated substrate 1-bromobutane was studied by means of 10 mM chromate , 0.1 mM cetyltrimethylammonium bromide (pH 9.2) in combination with indirect detection or 20 mM ,-alanine , hydrochloric acid (pH 3.5) in combination with direct detection. The method was used to estimate the inhibition constant KI (0.44 mM by indirect detection and 0.63 mM by of direct detection) and to determine the inhibition type. Compared to spectrophotometric and other discontinuous assays, the method is rapid, can be automated, and requires only small amount of reagents that is especially important in the case of enzymes and inhibitors. [source] Role of IscS in Fe-S cluster assembly in Trypanosoma bruceiTHE JOURNAL OF EUKARYOTIC MICROBIOLOGY, Issue 2 2005O. SMÍD Despite the significance of proteins containing iron-sulfur cluster (Fe,S proteins), the processes of Fe,S cluster assembly and maturation of Fe,S proteins are poorly understood. However, several key proteins involved in the assembly have been identified, notably IscS, a cystein desulfurase, which provides sulfur for Fe,S cluster and IscU, a metallochaperone acting as a scaffold for cluster assembly. In this work, we studied the process of Fe,S cluster biosynthesis in Trypanosoma brucei by identifying the homologue of IscS in the T. brucei (TbIscS). To address the function of TbIscS, we inhibited its expression by means of RNA interference (RNAi). After RNAi induction, generation time of the TbIscS knock-down cell line was significantly prolonged. All types of mitochondrial ATP production in the cells were severely affected. Analysis of glucose metabolism end products determined pyruvate as major excreted metabolite of the induced cells, while the uninduced cells produced only small amount of this glycolytic end product. These data demonstrate that mitochondrial metabolism is impaired in cells with TbIscS knocked down. To test whether the observed phenomena were results of Fe,S cluster assembly disruption, we examined the Fe,S cluster-dependent activity of aconitase. This enzyme is localized in its active form in mitochondrion as well as in cytosol of T. brucei. After RNAi induction we observed the reduction of aconitase activity in both compartments (approx. 70% reduction in cytosol, approx. 30% in mitochondria). Western blots together with the EPR analysis showed that the reduction in cytosolic activity was due to impaired Fe,S cluster formation, while decrease in aconitase activity in mitochondria corresponded to the reduced level of the protein. [source] Axoneme-dependent tubulin modifications in singlet microtubules of the Drosophila sperm tailCYTOSKELETON, Issue 4 2008Henry D. Hoyle Abstract Drosophila melanogaster sperm tubulins are posttranslationally glutamylated and glycylated. We show here that axonemes are the substrate for these tubulin C-terminal modifications. Axoneme architecture is required, but full length, motile axonemes are not necessary. Tubulin glutamylation occurs during or shortly after assembly into the axoneme; only glutamylated tubulins are glycylated. Tubulins in other testis microtubules are not modified. Only a small subset of total Drosophila sperm axoneme tubulins have these modifications. Biochemical fractionation of Drosophila sperm showed that central pair and accessory microtubules have the majority of poly-modified tubulins, whereas doublet microtubules have only small amounts of mono- and oligo-modified tubulins. Glutamylation patterns for different ,-tubulins experimentally assembled into axonemes were consistent with utilization of modification sites corresponding to those identified in other organisms, but surrounding sequence context was also important. We compared tubulin modifications in the 9 + 9 + 2 insect sperm tail axonemes of Drosophila with the canonical 9 + 2 axonemes of sperm of the sea urchin Lytichinus pictus and the 9 + 0 motile sperm axonemes of the eel Anguilla japonica. In contrast to Drosophila sperm, L. pictus sperm have equivalent levels of modified tubulins in both doublet and central pair microtubule fractions, whereas the doublets of A. japonica sperm exhibit little glutamylation but extensive glycylation. Tubulin C-terminal modifications are a prevalent feature of motile axonemes, but there is no conserved pattern for placement or amount of these modifications. We conclude their functions are likely species-specific. Cell Motil. Cytoskeleton 2008. © 2008 Wiley-Liss, Inc. [source] Focused proteomics: Monoclonal antibody-based isolation of the oxidative phosphorylation machinery and detection of phosphoproteins using a fluorescent phosphoprotein gel stainELECTROPHORESIS, Issue 15 2004James Murray Abstract We have raised monoclonal antibodies capable of immunocapturing all five complexes involved in oxidative phosphorylation for evaluating their post-translational modifications. Complex I (NADH dehydrogenase), complex II (succinate dehydrogenase), complex III (cytochrome c reductase), complex IV (cytochrome c oxidase), and complex V (F1F0 ATP synthase) from bovine heart mitochondria were obtained in good yield from small amounts of tissue in more than 90% purity in one step. The composition and purity of the complexes was evaluated by Western blotting using monoclonal antibodies against individual subunits of the five complexes. In this first study, the phosphorylation state of the proteins without inducing phosphorylation or dephosphorylation was identified by using the novel Pro-Q Diamond phosphoprotein gel stain. The major phosphorylated components were the same as described before in sucrose gradient enriched complexes. In addition a few additional potential phosphoproteins were observed. Since the described monoclonal antibodies show cross reactivity to human proteins, this procedure will be a fast and efficient way of studying post-translational modifications in control and patient samples using only small amounts of tissue. [source] Carbon and nitrogen isotope composition of bulk soils, particle-size fractions and organic material after treatment with hydrofluoric acidEUROPEAN JOURNAL OF SOIL SCIENCE, Issue 3 2005M. W. I. Schmidt Summary Soils and sediments contain only small amounts of organic matter, and large concentrations of paramagnetic metals can give poor solid-state nuclear magnetic resonance (NMR) spectra of organic matter. Pretreatment of samples with hydrofluoric acid (HF) dissolves significant proportions of the mineral matrix and extracts paramagnetic elements. We investigated the effects of 10% HF treatment on the stable isotope content of carbon (C) and nitrogen (N) of organic matter from soils, composts and shales. Additionally we inferred molecular and isotopic characteristics of lost materials from calculations of isotope mass balances. Treatment with HF enriched C and N in mineral samples substantially (factors 2.5,42.4), except for Podzol B horizons (1.1,1.7) and organic material (1.0,1.3). After treatment most of the C (59.7,91.7%) and N (53.7,86.6%) was recovered, although changing C/N ratios often indicated a preferential loss of N-rich material. Isotope ratios of C and N in the remaining material became more negative when net alterations exceeded 0.3,. The isotope ratios of the lost material contained more 13C (1,2,) and 15N (1,4,) than the initial organic matter. Acid hydrolysis typically removes proteins, amino acids and polysaccharides, all of which are enriched in 13C, and in the case of proteins and amino acids, enriched in 15N as well. We conclude that HF treatment released fresh, soluble, probably microbial, biomass in addition to carbohydrates. Net changes of the bulk chemical composition of organic matter were small for most soils, size fractions and plant material, but not for samples containing little organic matter, or those rich in easily soluble organic matter associated with iron oxides, such as Podzol B horizons. [source] Protein array technology to investigate cytokine production by monocytes from patients with advanced alcoholic cirrhosis: An ex vivo pilot studyHEPATOLOGY RESEARCH, Issue 7 2009Khalid A. Tazi Aim:, In patients with advanced cirrhosis, little is known about the ability of peripheral blood monocytes to spontaneously produce signaling proteins such as cytokines. The aim of this ex vivo study was to evaluate cytokine production under baseline conditions and after stimulation by lipopolysaccharide (LPS), a toll-like receptor (TLR) agonist. Methods:, Peripheral blood monocytes were isolated from patients with advanced alcoholic cirrhosis (without ongoing bacterial infections) and normal subjects. Cells were left unstimulated or were stimulated with LPS. The abundance of 24 cytokines was measured using a filter-based, arrayed sandwich enzyme-linked immunosorbent assay (ELISA) in the supernatant of cultured monocytes. Results:, Cirrhotic monocytes spontaneously produced six proteins (TNF-,, IL-6, IL-8, MCP-1, RANTES and Gro), whereas normal monocytes produced only small amounts of IL-8 and RANTES. Analyses with the online gene set analysis toolkit WebGestalt (http://bioinfo.vanderbilt.edu/webgestalt) found enrichment for the six proteins in the human gene ontology subcategory (http://www.geneontology.org), Kyoto Encyclopedia of Genes and Genome pathways (http://www.genome.ad.jp/kegg/) and BioCarta pathways (http://www.biocarta.com/genes/index.asp) consistent with a proinflammatory phenotype of cirrhotic monocytes resulting from activated TLR signaling. Interestingly, LPS-elicited TLR engagement further increased the production of the six proteins and did not induce the secretion of any others, in particular the anti-inflammatory cytokine IL-10. LPS-stimulated normal monocytes produced TNF-,, IL-6, IL-8, MCP-1, RANTES, Gro and IL-10. Conclusion:, In patients with advanced cirrhosis, peripheral blood monocytes spontaneously produce proinflammatory cytokines, presumably in response to unrestricted TLR signaling. [source] DNA Extraction from Olive Oil and PCR Amplification of Microsatellite MarkersJOURNAL OF FOOD SCIENCE, Issue 1 2005Raffaele Testolin And ABSTRACT: DNA was extracted from single-cultivar of cold-pressed (virgin) unfiltered and cotton-filtered olive oils that were stored at 4 °C for up to a year using different DNA extraction kits and protocols. DNA was amplified using original and nested primers designed on 6 microsatellites loci of the UDO series. The most consistent results in terms of successful single sequence repeat amplifications were achieved using the Qiagen QIAamp DNA stool extraction kit, slightly modified and applied to oil sample amounts as small as 200 ,L without any pretreatment. The kit allowed getting polymerase chain reaction (PCR) amplicons visible on gel and scorable peaks at the automatic sequencer for all 6 markers analyzed. Less consistent results were achieved with other kits, such as the Promega Wizard Magnetic DNA Purification System for Food, the LB Link-Biotech ExtMan 50,100 Evolution, the Qiagen Plant Mini kit, and the standard cetyltrimethyl-ammonium bromide-based DNA extraction protocol. The integration in the protocols of further tools, such as the hexane-based phase separation, the addition of water or NaCl solutions to the oil, the precipitation and the use of the pellet, and others, did not result in any substantial use. PCR amplifications that gave low DNA yields were improved by adopting the nested PCR technique, which uses the product of the 1st PCR as a template for a 2nd PCR carried out by means of internal primers. Conclusions are drawn as to the applicability of the method to trace the identity of single-cultivar virgin olive oils. Further work is required to check the sensitivity of the method in determining the varietal composition of blended oils, especially in detecting alleles from cultivars present in only small amounts. [source] Adult large pine weevils Hylobius abietis feed on silver birch Betula pendula even in the presence of conifer seedlingsAGRICULTURAL AND FOREST ENTOMOLOGY, Issue 2 2006Riitta Toivonen Abstract 1,The feeding preference of the adult pine weevil Hylobius abietis (L.) (Coleoptera: Curculionidae) for Betula pendula Roth was studied in no-choice and paired-choice feeding experiments. 2,In the first no-choice test, large quantities of silver birch bark in Petri dishes were consumed; on average, the daily consumption of each weevil was 67 mm2. 3,In the second no-choice test, the weevils were offered 1-year-old silver birch seedlings for 6 days. Initially, the weevils fed mostly on the stem bases; later, they moved upward to feed on other parts of the stems. In addition to the main shoots, scars caused by gnawing were also found on leaf bases, blades, veins and petioles. Feeding resulted in the death of the main stems in 15% of the seedlings. 4,In the paired-choice tests, the conifers were preferred to silver birch, even though a large amount of silver birch was also consumed in the presence of conifers. 5,In the paired-choice tests, equal amounts of Scots pine and Norway spruce were always consumed. When hybrid aspen was offered, only small amounts were gnawed. [source] Mapping the characteristics of the radical ring-opening polymerization of a cyclic ketene acetal towards the creation of a functionalized polyesterJOURNAL OF POLYMER SCIENCE (IN TWO SECTIONS), Issue 18 2009Peter Plikk Abstract Radical ring-opening polymerization of cyclic ketene acetals is a means to achieve novel types of aliphatic polyesters. 2-methylene-1,3-dioxe-5-pene is a seven-membered cyclic ketene acetal containing an unsaturation in the 5-position in the ring structure. The double bond functionality enables further reactions subsequent to polymerization. The monomer 2-methylene-1,3-dioxe-5-pene was synthesized and polymerized in bulk by free radical polymerization at different temperatures, to determine the structure of the products and propose a reaction mechanism. The reaction mechanism is dependent on the reaction temperature. At higher temperatures, ring-opening takes place to a great extent followed by a new cyclization process to form the stable five-membered cyclic ester 3-vinyl-1,4-butyrolactone as the main reaction product. Thereby, propagation is suppressed and only small amounts of other oligomeric products are formed. At lower temperatures, the cyclic ester formation is reduced and oligomeric products containing both ring-opened and ring-retained repeating units are produced at higher yield. © 2009 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 47: 4587,4601, 2009 [source] Preliminary study on the monitoring of glutathione S-tranferase activity toward styrene oxide by electromigration methodsJOURNAL OF SEPARATION SCIENCE, JSS, Issue 12 2005ková Abstract A new method has been developed for the monitoring of glutathione S-tranferase (GST) detoxification activity toward styrene oxide (SO). The enzymatic reaction was carried out directly in a thermostatted autosampler vial and the formation of conjugates between glutathione (GSH) and SO was monitored by sequential MEKC runs. The determinations were performed in a 50-,m fused silica capillary using 50 mM SDS in 20 mM phosphate 20 mM tetraborate buffer (pH 8.3) as a background electrolyte; separation voltage 28 kV (positive polarity), temperature of capillary 25°C, and detection at 200 nm. The method is rapid, amenable to automation, and requires only small amounts of samples, which is especially important in the case of GST isoenzyme analyses. [source] Effect of melt processing conditions on the morphology and properties of nylon 6 nanocompositesPOLYMER ENGINEERING & SCIENCE, Issue 11 2007F. Chavarria Nylon 6 (PA-6) organoclay nanocomposites were prepared by melt processing using three different twin screw extruders (TSEs). The effect of mixing conditions, feed port location, residence time, and number of extrusion passes on the morphology and mechanical properties of the nanocomposites were examined. Wide-angle X-ray scattering, transmission electron microscopy (TEM), and mechanical property data are reported. Particle analyses were performed on the TEM images to quantitatively characterize the extent of exfoliation. The amount of shear and the mixing conditions created by TSEs have a significant effect on the morphology and properties of PA-6 nanocomposites. Morphology and mechanical property results show that (1) melting the polymer before coming into contact with the organoclay followed by a low level of shear and (2) maintaining a medium level of shear throughout the extruder with a longer residence time lead to extremely high platelet dispersion and matrix reinforcement for PA-6 nanocomposites. Nanocomposites formed in a DSM microcompounder showed similar morphologies and modulus trends as those obtained with conventional TSEs; thus, this microcompounder is a good alternative for nanocomposite research especially when only small amounts of material are available. POLYM. ENG. SCI., 47:1847,1864, 2007. © 2007 Society of Plastics Engineers [source] An Arabidopsis porB porC double mutant lacking light-dependent NADPH:protochlorophyllide oxidoreductases B and C is highly chlorophyll-deficient and developmentally arrestedTHE PLANT JOURNAL, Issue 2 2003Geneviève Frick Summary A key reaction in the biosynthesis of chlorophylls (Chls) a and b from cyanobacteria through higher plants is the strictly light-dependent reduction of protochlorophyllide (Pchlide) a to chlorophyllide (Chlide) a. Angiosperms, unlike other photosynthetic organisms, rely exclusively upon this mechanism to reduce Pchlide and hence require light to green. In Arabidopsis, light-dependent Pchlide reduction is mediated by three structurally related but differentially regulated NADPH:Pchlide oxidoreductases, denoted as PORA, PORB, and PORC. The PORA and PORB genes, but not PORC, are strongly expressed early in seedling development. In contrast, expression of PORB and PORC, but not PORA, is observed in older seedlings and adult plants. We have tested the hypothesis that PORB and PORC govern light-dependent Chl biosynthesis throughout most of the plant development by identifying porB and porC mutants of Arabidopsis, the first higher plant por mutants characterized. The porB-1 and porC-1 mutants lack the respective POR transcripts and specific POR isoforms because of the interruption of the corresponding genes by a derivative of the maize Dissociation (Ds) transposable element. Single por mutants, grown photoperiodically, display no obvious phenotypes at the whole plant or chloroplast ultrastructural levels, although the porB-1 mutant has less extensive etioplast inner membranes. However, a light-grown porB-1 porC-1 double mutant develops a seedling-lethal xantha phenotype at the cotyledon stage, contains only small amounts of Chl a, and possesses chloroplasts with mostly unstacked thylakoid membranes. PORB and PORC thus seem to play redundant roles in maintaining light-dependent Chl biosynthesis in green plants, and are together essential for growth and development. [source] Structural and Histochemical Studies on the Teleostean Bulbus ArteriosusANATOMIA, HISTOLOGIA, EMBRYOLOGIA, Issue 6 2009I. L. Leknes Summary The structure and histochemical properties of the bulbus arteriosus in two species from an evolutionary old teleost family, Characidae, and in three modern teleosts, family Cichlidae, are described. The bulbar wall was composed of an outer layer, a middle layer and a strongly folded inner layer covered by a thick, granule-rich endothelial cell layer towards the lumen. One of the cichlid species (Thorichthys meeki) was injected intraperitoneally with horse ferritin; the endothelial cell layer of the heart atrium and ventricle displayed high ability to endocytose ferritin particles from the blood stream, but the corresponding layer in the bulbus arteriosus displayed no such uptake. This finding suggests that the bulbar endothelial cell layer plays no scavenger or immunological blood cleansing roles in this species. The bulbar endothelial cell granules were strongly coloured by periodic acid,Schiff (PAS) in the present cichlids, but weakly coloured by PAS in the present characids. These cell layers were uncoloured by alkaline carmine in ethanol in both cichlids and characids. The negative carmine test combined with a positive PAS test for the bulbar endothelial cell layer in the present cichlids indicates that these cells contain only small amounts of polysaccharides. The weak PAS-colouring for the bulbar endothelial cell layer in characids indicates a very low content of sugars in these cells. These findings together with the fact that this cell layer in the present cichlids and characids was nearly uncoloured when treated with orcein, Heidenhain's Azan or Schmorl's solutions for elastic materials suggest that the bulbar endothelial granules do not play any role in the blood cleansing or in the rebuilding or maintenance of the ground substance or elastic material in the bulbar wall. Probably, the granules in the bulbar endothelial cell layer in the present species contain mainly proteins, connected to some PAS-positive polysaccharides to enhance their solubility. [source] Organoarsenic compounds in plants and soil on top of an ore veinAPPLIED ORGANOMETALLIC CHEMISTRY, Issue 5 2002Anita Geiszinger Abstract Plants and soil collected above an ore vein in Gasen (Austria) were investigated for total arsenic concentrations by inductively coupled plasma mass spectrometry (ICP-MS). Total arsenic concentrations in all samples were higher than those usually found at non-contaminated sites. The arsenic concentration in the soil ranged from ,700 to ,4000,mg kg,1 dry mass. Arsenic concentrations in plant samples ranged from ,0.5 to 6,mg kg,1 dry mass and varied with plant species and plant part. Examination of plant and soil extracts by high-performance liquid chromatography,ICP-MS revealed that only small amounts of arsenic (<1%) could be extracted from the soil and the main part of the extractable arsenic from soil was inorganic arsenic, dominated by arsenate. Trimethylarsine oxide and arsenobetaine were also detected as minor compounds in soil. The extracts of the plants (Trifolium pratense, Dactylis glomerata, and Plantago lanceolata) contained arsenate, arsenite, methylarsonic acid, dimethylarsinic acid, trimethylarsine oxide, the tetramethylarsonium ion, arsenobetaine, and arsenocholine (2.5,12% extraction efficiency). The arsenic compounds and their concentrations differed with plant species. The extracts of D. glomerata and P. lanceolata contained mainly inorganic arsenic compounds typical of most other plants. T. pratense, on the other hand, contained mainly organic arsenicals and the major compound was methylarsonic acid. Copyright © 2002 John Wiley & Sons, Ltd. [source] Comparative analysis of two biliproteins, BP1 and BP2, from haemolymph of cabbage white butterfly, Pieris rapaeARCHIVES OF INSECT BIOCHEMISTRY AND PHYSIOLOGY (ELECTRONIC), Issue 4 2006Chi Won Choi Abstract Two blue-pigment binding proteins, BP1 and BP2, are present in larval and pupal haemolymph of cabbage white butterfly, Pieris rapae, and fluctuate in expression during development. Both BP1 and BP2 are found in pupal haemolymph in varying proportions as well as in adult haemolymph, while only small amounts of BP2 are found in larval haemolymph. BPs are separated by 75% ammonium sulfate, and then purified effectively by ion exchange column chromatography and preparative gel electrophoresis. It was shown that BP1 and BP2 have molecular masses of 20,244 and 19,878 Da, and isoelectric points of 7.0 and 6.8, respectively. Considering their amino acid compositions and N-terminal amino acid sequences, the two proteins are almost identical except the first N-terminal amino acid. The first amino acid of BP1 is asparagine, whereas the initial residue of BP2 is aspartic acid. Anti-BP1 cross-reacts with BP2, indicating that they have immunological homogeneity. Western blotting analyses revealed that only BP1 was present in the larval tissues such as fat body, integument, muscle, and hindgut. However, BP1 was not found in midgut, Malphigian tubules, and silk gland. BP1 was also present in the protein bodies, and both cuticle and hemocoel sides of larval epidermis cells by the transmission electron microscopic observation. The information in this report will facilitate studies on the molecular biology and biological significance of insect BPs. Arch. Insect Biochem. Physiol. 61:220,230, 2006. © 2006 Wiley-Liss, Inc. [source] Effects of hydrophobicity and anions on self-assembly of the peptide EMK16-IIBIOPOLYMERS, Issue 4 2010Dawei Zou Abstract Effects of hydrophobic and electrostatic interactions on the self-assembling process of the ionic-complementary peptide EMK16-II are investigated by atomic force microscopy imaging, circular dichroism spectra, light scattering, and chromatography. It is found that the hydrophobicity of the peptide promotes the aggregation in pure water even at a very low concentration, resulting in a much lower critical aggregation concentration than that of another peptide, EAK16-II. The effect of anions in solution with different valences on electrostatic interactions is also important. Monovalent anions (Cl, and Ac,) with a proper concentration can facilitate the formation of peptide fibrils, with Cl, of smaller size being more effective than Ac, of larger size. However, only small amounts of fibrils, but plenty of large amorphous aggregates, are found when the peptide solution is incubated with multivalent anions, such as SO, C6H5O, and HPO. More importantly, by gel filtration chromatography, the citrate anion, which induces a similar effect on the self-assembling process of EMK16-II as that of SO and HPO, can interact with two or more positively charged residues of the peptide and reside in the amorphous aggregates. This implies a "salt bridge" effect of multivalent anions on the peptide self-assembling process, which can interpret a previous puzzle why divalent cations inhibit the formation of ordered nanofibrils of the ionic-complementary peptides. Thus, our results clarify the important effects of hydrophobic and electrostatic interactions on the self-assembling process of the ionic-complementary peptides. These are greatly helpful for us to understand the mechanism of peptides' self-assembling process and protein folding and aggregation. © 2009 Wiley Periodicals, Inc. Biopolymers 93: 318,329, 2010. This article was originally published online as an acceptedpreprint. The "Published Online" date corresponds to the preprintversion. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com [source] Effect of Different Carbon Sources on the Production of Succinic Acid Using Metabolically Engineered Escherichia coliBIOTECHNOLOGY PROGRESS, Issue 2 2007Christian Andersson Succinic acid (SA) is an important platform molecule in the synthesis of a number of commodity and specialty chemicals. In the present work, dual-phase batch fermentations with the E. coli strain AFP184 were performed using a medium suited for large-scale industrial production of SA. The ability of the strain to ferment different sugars was investigated. The sugars studied were sucrose, glucose, fructose, xylose, and equal mixtures of glucose and fructose and glucose and xylose at a total initial sugar concentration of 100 g L,1. AFP184 was able to utilize all sugars and sugar combinations except sucrose for biomass generation and succinate production. For sucrose as a substrate no succinic acid was produced and none of the sucrose was metabolized. The succinic acid yield from glucose (0.83 g succinic acid per gram glucose consumed anaerobically) was higher than the yield from fructose (0.66 g g,1). When using xylose as a carbon source, a yield of 0.50 g g,1 was obtained. In the mixed-sugar fermentations no catabolite repression was detected. Mixtures of glucose and xylose resulted in higher yields (0.60 g g,1) than use of xylose alone. Fermenting glucose mixed with fructose gave a lower yield (0.58 g g,1) than fructose used as the sole carbon source. The reason is an increased pyruvate production. The pyruvate concentration decreased later in the fermentation. Final succinic acid concentrations were in the range of 25,40 g L,1. Acetic and pyruvic acid were the only other products detected and accumulated to concentrations of 2.7,6.7 and 0,2.7 g L,1. Production of succinic acid decreased when organic acid concentrations reached approximately 30 g L,1. This study demonstrates that E. coli strain AFP184 is able to produce succinic acid in a low cost medium from a variety of sugars with only small amounts of byproducts formed. [source] Hydrogenogenic CO Conversion in a Moderately Thermophilic (55 °C) Sulfate-Fed Gas Lift Reactor: Competition for CO-Derived H2BIOTECHNOLOGY PROGRESS, Issue 5 2006Jan Sipma Thermophilic (55 °C) sulfate reduction in a gas lift reactor fed with CO gas as the sole electron donor was investigated. The reactor was inoculated with mesophilic granular sludge with a high activity of CO conversion to hydrogen and carbon dioxide at 55 °C. Strong competition for H2 was observed between methanogens and sulfate reducers, while the homoacetogens present consumed only small amounts of H2. The methanogens appeared to be more sensitive to pH and temperature shocks imposed to the reactor, but could not be completely eliminated. The fast growth rates of the methanogens (generation time of 4.5 h) enabled them to recover fast from shocks, and they rapidly consumed more than 90% of the CO-derived H2. Nevertheless, steep increases in sulfide production in periods with low methane production suggests that once methanogenesis is eliminated, sulfate reduction with CO-rich gas as electron donor has great potential for thermophilic biodesulfurization. [source] Pathways and Substrate Specificity of DMSP Catabolism in Marine Bacteria of the Roseobacter CladeCHEMBIOCHEM, Issue 3 2010Jeroen S. Dickschat Dr. Abstract The volatiles released by Phaeobacter gallaeciensis, Oceanibulbus indolifex and Dinoroseobacter shibae have been investigated by GC-MS, and several MeSH-derived sulfur volatiles have been identified. An important sulfur source in the oceans is the algal metabolite dimethylsulfoniopropionate (DMSP). Labelled [2H6]DMSP was fed to the bacteria to investigate the production of volatiles from this compound through the lysis pathway to [2H6]dimethylsulfide or the demethylation pathway to [2H3]-3-(methylmercapto)propionic acid and lysis to [2H3]MeSH. [2H6]DMSP was efficiently converted to [2H3]MeSH by all three species. Several DMSP derivatives were synthesised and used in feeding experiments. Strong dealkylation activity was observed for the methylated ethyl methyl sulfoniopropionate and dimethylseleniopropionate, as indicated by the formation of EtSH- and MeSeH-derived volatiles, whereas no volatiles were formed from dimethyltelluriopropionate. In contrast, the dealkylation activity for diethylsulfoniopropionate was strongly reduced, resulting in only small amounts of EtSH-derived volatiles accompanied by diethyl sulfide in P. gallaeciensis and O. indolifex, while D. shibae produced the related oxidation product diethyl sulfone. The formation of diethyl sulfide and diethyl sulfone requires the lysis pathway, which is not active for [2H6]DMSP. These observations can be explained by a shifted distribution between the two competing pathways due to a blocked dealkylation of ethylated substrates. [source] Hochdurchsatz-Methoden in der Festkörperchemie.CHEMIE IN UNSERER ZEIT (CHIUZ), Issue 5 2007Schneller zum Ziel Abstract Hochdurchsatz-Methoden zeichnen sich gegenüber konventionellen Methoden durch eine größere Effizienz aus. Mittlerweile steht im Bereich der Anorganischen Festkörperchemie und der Materialwissenschaften eine Vielzahl von Werkzeugen zur Verfügung, um die Entdeckung und Optimierung von Materialien durch Verwendung von HD-Methoden schneller voranzutreiben. Für alle Arbeitstechniken gilt: Das Ausmaß der Automatisierung, Parallelisierung und Miniaturisierung sowie die Integration aller Einzelschritte in einen Gesamtprozess bestimmt die Effizienz einer HD-Methodik. Durch eine systematische Untersuchung des Parameterraumes und durch sorgfältige Datenevaluierung (möglichst Software-gestützt) können Trends gefunden werden, die zu einem besseren Verständnis der untersuchten Systeme beitragen. Der vorgestellte Ansatz zur HD-Untersuchung von Solvothermalreaktionen erlaubt die automatisierte Synthese und Charakterisierung von bis zu 48 Produkten ohne die Proben einzeln manipulieren zu müssen. Der geringe Reagenzienverbrauch zusammen mit der Möglichkeit, die Reaktionen unter identischen Prozessparametern (Temperatur, Zeit, Druck, usw.) durchführen zu können, ermöglichen die systematische Untersuchung eines größeren Parameterraumes. An Hand zahlreicher Beispiele haben wir das Potential von Hochdurchsatz- Methoden bei der Entdeckung neuer Verbindungen, der Optimierung von Reaktionen, beim Auffinden von Reaktionstrends sowie der Untersuchung des Einflusses von Prozessparametern gezeigt. High-throughput (HT) methods applied to materials discovery have attracted much attention over the last few years because they permit a fast and efficient investigation of parameter space while consuming only small amounts of starting materials. Nowadays, many different HT techniques in solid state science are employed in order to accelerate the discovery and optimization of materials. The degree of parallelisation, automation, and miniaturization determines the efficiency of a HT methodology. Systematic investigation of parameter space and careful data analysis allow the identification of reaction trends and may give important guidance in better understanding of reaction systems. The described HT methodology for solvothermal reactions allows the systematic investigation of 48 different hydrothermal reactions at a time. The methodology includes automatic dispensing of solids and liquids, followed by homogenization, pH measurement, synthesis, isolation, washing as well as automated phase analysis by X-ray diffraction without the manipulation of individual samples. The focus of this article is on the application of HT methods in the investigation of the parameter space in solid state sciences. Based on selected examples, the power of HT methods in the discovery of new compounds, the optimization of reactions, the identification of reaction trends, as well as the investigation of the influence of process parameters is described. The large amount of data obtained in a short time leads to an improvement towards the understanding of the role that synthesis and process parameters play in the formation of compounds and materials. [source] Histologic Analysis of Clinical Biopsies Taken 6 Months and 3 Years after Maxillary Sinus Floor Augmentation with 80% Bovine Hydroxyapatite and 20% Autogenous Bone Mixed with Fibrin GlueCLINICAL IMPLANT DENTISTRY AND RELATED RESEARCH, Issue 2 2001Mats Hallman DDS Abstract: Background: Bovine hydroxyapatite (Bio-Oss®, Geistlich Pharmaceutical, Wollhausen, Switzerland) has been suggested to be used in maxillary sinus floor augmentation procedures prior to or in conjunction with implant placement. However, the long-term histologic fate of this material is not well understood. Purpose: The aim with this study was to histologically evaluate the tissue response in patients to a mixture of bovine hydroxyapatite (BH), autogenous bone, and fibrin glue 6 months and 3 years after a maxillary sinus floor augmentation procedure. Materials and Method: Biopsies were taken from a group of 20 consecutive patients 6 months (n = 16) and 3 years (n = 12) after maxillary sinus floor augmentation with a mixture of BH (80%), autogenous bone (20%), and fibrin glue and prepared for histologic analysis. Results: Light microscopy and morphometry from biopsies taken after 6 months showed various amounts of mineralized bone tissue. The specimen area was occupied by 54.1 ± 12.6% nonmineralized tissue, followed by 21.2 ± 24.5% lamellar bone, 14.5 ± 10.3% BH particles, and 10.2 ± 13.4% woven bone. The nonmineralized tissue seen in bone-forming areas consisted of a loose connective tissue, rich with vessels and cells. There were no signs of resorption of the BH particles. The lamellar bone appeared to have originated from the recipient site and was seldom in contact with the BH particles. After 3 years, the nonmineralized tissue area had decreased to 36.0 ± 19.0% (p > .05) and consisted mainly of bone marrow tissue. The surface area of lamellar bone had increased to 50.7 ± 22.8% (p > .05), and there was almost no immature bone. The mean specimen area occupied by BH particles, was 12.4 ± 8.7% and had not changed from 6 months (not significant). Moreover, the sizes of the particles were similar after 6 months and 3 years. The degree of BH particle,bone contact had increased from 28.8%± 19.9% after 6 months to 54.5 ± 28.8% after 3 years (p > .05). Conclusion: Histology of specimens from maxillary sinuses augmented with 80% BH particles, 20% autogenous bone, and fibrin glue showed a positive bone tissue response after 6 months and 3 years after augmentation of the maxillary sinus floor prior to implant placement in a group fo 20 patients. The bone surrounding and in contact with the BH particles after 6 months was mainly immature woven bone, which with time was replaced by mature lamellar bone filling the interparticle space as observed in the 3-year specimens. Moreover, bone-integrated BH particles seem to be resistant to resorption. The results indicate that the procedure may be considered when only small amounts of intraoral autogenous bone graft are available. [source] | ||||||||||||||||||||||