Properties of a single-chain antibody containing different linker peptides

Single-chain antibodies were constructed using six differentlinker peptides to join the V_H and V_L domains of an anti-2-phenyloxazolone(Ox) antibody. Four of the linker peptides originated from theinterdomain linker region of the fungal cellulase CBHI and consistedof 28, 11, six and two amino acid residues. The two other linkerpeptides used were the (GGGGS)₃ linker with 15 amino acid residuesand a modified IgG2b hinge peptide with 22 residues. Proteolyticstability and Ox binding properties of the six different scFvderivatives produced in Escherichia coli were investigated andcompared with those of the corresponding Fv fragment containingno joining peptide between the V domains. The hapten bindingproperties of different antibody fragments were studied by ELISAand BIAcore^TM. The interdomain linker peptide improved the haptenbinding properties of the antibody fragment when compared withFv fragment, but slightly increased its susceptibility to proteases.Single-chain antibodies with short CBHI linkers of 11, six andtwo residues had a tendency to form multimers which led to ahigher apparent affinity. The fragments with linkers longerthan 11 residues remained monomeric.     

Introduction of lysine residues on the light chain constant domain improves the labelling properties of a recombinant Fab fragment

Europium chelates provide a non-radioactive alternative forsensitive labelling of antibodies for diagnostic immunoassays.Lysine residues at antibody surfaces are ready targets for labellingby an isothiocyanate derivative of the europium chelate (Eu³⁺).Here the labelling efficiency of a recombinant anti-human -fetoprotein(hAFP) Fab fragment has been improved by increasing its lysinecontent by protein engineering. Molecular modelling was usedto identify three light chain constant domain surface arginineresidues, R154, R187 and R210, which were mutated to lysineresidues. The mutations did not influence the affinity of thelysine-enriched Fab fragment and its labelling efficiency wasfound to be 40% higher than that of the wildtype Fab fragmentWith low degree of labelling, the affinities of the two Fabfragments were identical and comparable with that of the originalmonoclonal anti-hAFP IgG. With a higher degree of labellingthe affinities of both Fab fragments decreased more than thatof the intact IgG since more lysine residues are available forlabelling in the additional heavy chain constant domains ofthe larger molecule. Electrostatic adsorption and covalent immobilizationof the Fab fragments were characterized by BIAcore^TM and thelysine-enriched Fab fragment was found to be more efficientlyimmobilized to an activated carboxymethyl surface.     

Volatile monocarbonyl compounds of carrot roots at various stages of maturity

Summary Volatile carbonyl compounds of carrot varietyFeonia Hunderup S-64 at various stages of maturity were analysed as their 2,4-DNPH's by a combined TLC-GLC-MS method. Twenty-three different carbonyl compounds were identified, of which undecanal, buten-2-al, methylbutenal, pentan-2-one, 6-methyl-5-hepten-2-one and 5-methylfurfural have not previously been found in carrot. During maturation the content of acetaldehyde and acetone increased significantly, while that of methylbutenal decreased.

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Die leichtflüchtigen Monocarbonylverbindungen der Karotten in verschiedenen Stufen der Reife

Zusammenfassung Die leichtflüchtigen Carbonylverbindungen der Karotten Varietät Feonia Hunderup S-64 wurden in verschiedenen Stufen der Reife als 2,4-DNPH mit der kombinierten DC-GC-MS-Methode analysiert. Dreiundzwanzig verschiedene Carbonylverbindungen wurden nachgewiesen. Von diesen sind Undecanal, 2-Butenal, Methylbutenal, 2-Pentanon, 6-Methyl-5-hepten-2-on und 5-Methylfurfural noch nicht in Karotten aufgefunden worden. Während des Reifens nahm der Gehalt an Acetaldehyd und Aceton bedeutend zu. Der Gehalt an Methylbutenal verminderte sich.

     

Changes in soil carbon stock after cropland conversion to grassland in Russian temperate zone: measurements versus model simulation

The collapse of Soviet Union in early 1990s led to abandonment of large area of arable land which is assumed to act as a carbon (C) sink. We studied the ability of two dynamic soil C models (Yasso07 and RothC) to predict changes in soil C content after cropland abandonment. The performance of the models was compared using the results of a long-term experiment in Pushchino, Moscow region (54°50′N, 37°35′E) in Russia. The experiment was divided in four combinations of fertilizer or mowing treatments on former cropland soil. The soil C content was determined in the year of establishment (1980) and thereafter in 1999 and 2004. The soil C stocks increased by about 1.5- to 1.8-fold during the study period. Both models predicted the overall change in soil C relatively well (modelling efficiency of Yasso07 and RothC were 0.60 and 0.73, respectively). According to the models, the soil gained on average 140–150 g C m^?2 year^?1 during the first 5 years after conversion of cropland to grassland. The C sequestration rate decreased to 40–50 g C m^?2 year^?1 after 20 years of land use change. The sequestration rates estimated in this study are comparable to the rates observed in other studies.     

Impact toughness,viscoelastic behavior,and morphology of polypropylene–jute–viscose hybrid composites

In this investigation, we studied the impact toughness and viscoelastic behavior of polypropylene (PP)–jute composites. In this study, we used viscose fiber as an impact modifier and maleated PP as a compatibilizer. The toughness of the composites was studied with conventional Charpy and instrumental falling‐weight impact tests. The composites’ viscoelastic properties were studied with dynamic mechanical analysis. The results show that the incorporation of viscose fibers improved the impact strength and toughness to 134 and 65% compared to those of the PP–jute composites. The tan δ peak amplitude also increased with the addition of the impact modifier and indicated a greater degree of molecular mobility. The thermal stability of the composites was evaluated with thermogravimetric analysis. The addition of 2 wt % maleated polypropylene (MAPP) to the impact‐modified composite improved the impact strength and toughness to 144 and 93%, respectively. The fiber–matrix morphology of the fracture surface and the Fourier transform infrared spectra were also studied to ascertain the existence of the type of interfacial bonds. Microstructural analysis showed the retention of viscose fibers in the composites compared to the more separated jute fibers. © 2015 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2016 , 133, 42981.     

Bio-methanol: How energy choices in the western United States can help mitigate global climate change

Converting available biomass from municipal, agricultural and forest wastes to bio-methanol can result in significant environmental and economic benefits. Keeping these benefits in mind, one plausible scenario discussed here is the potential to produce energy using bio-methanol in five states of the western United States. In this scenario, the bio-methanol produced is from different biomass sources and used as a substitute for fossil fuels in energy production. In the U.S. West, forest materials are the dominant biomass waste source in Idaho, Montana, Oregon and Washington, while in California, the greatest amount of available biomass is from municipal wastes. Using a 100% rate of substitution, bio-methanol produced from these sources can replace an amount equivalent to most or all of the gasoline consumed by motor vehicles in each state. In contrast, when bio-methanol powered fuel cells are used to produce electricity, it is possible to generate 12–25% of the total electricity consumed annually in these five states.As a gasoline substitute, bio-methanol can optimally reduce vehicle C emissions by 2–29 Tg of C (23–81% of the total emitted by each state). Alternatively, if bio-methanol supported fuel cells are used to generate electricity, from 2 to 32 Tg of C emissions can be avoided. The emissions avoided, in this case, could equate to 25–32% of the total emissions produced by these particular western states when fossil fuels are used to generate electricity. The actual C emissions avoided will be lower than the estimates here because C emissions from the methanol production processes are not included; however, such emissions are expected to be relatively low. In general, there is less carbon emitted when bio-methanol is used to generate electricity with fuel cells than when it is used as a motor vehicle fuel.In the state of Washington, thinning “high-fire-risk” small stems, namely 5.1–22.9 cm diameter trees, from wildfire-prone forests and using them to produce methanol for electricity generation with fuel cells would avoid C emissions of 3.7–7.3 Mg C/ha. Alternatively, when wood-methanol produced from the high-fire-risk wood is used as a gasoline substitute, 3.3–6.6 Mg C/ha of carbon emissions are avoided. If these same “high-fire-risk” woody stems were burned during a wildfire 7.9 Mg C/ha would be emitted in the state of Washington alone. Although detailed economic analyses of producing methanol from biomass are in its infancy, we believe that converting biomass into methanol and substituting it for fossil-fuel-based energy production is a viable option in locations that have high biomass availability.     

Surface modification of Scots pine: the effect of process parameters on the through thickness density profile

This study evaluated the significance of different process parameters (press temperature, closing time, holding time, moisture content and compression ratio) on solid wood surface densification and its effect on the density profile generated in Scots pine sapwood. Changes in the microstructure of the wood were also evaluated microscopically. The results showed that with a shorter closing time, densification occurred closer to the sample surface than with an extended closing time. At a compression temperature of 150 °C, the vertical density profile exhibited a sharp peak in density that was close to the wood surface. A higher temperature of 200 °C resulted in a slightly broader density peak that was less intense and further from the surface. A holding time of 10 min resulted in the wood compressing to a slightly greater extent than when using a holding time of 1 min. Higher moisture content led to more extensive deformation. The results indicate that surface modification by densification is a viable method of enhancing wood properties.     

Effect of Trichoderma reesei tyrosinase on rheology and microstructure of acidified milk gels

The aim of this work was to study the potential of tyrosinase enzymes in structural engineering of acid-induced milk protein gels. Fat free raw milk, heated milk or a sodium caseinate solution were treated with tyrosinases from Trichoderma reesei (TrTyr) and Agaricus bisporus (AbTyr) and the reference enzyme transglutaminase (TG) prior to acid-induced gelation. TrTyr treatment increased the firmness of raw milk and sodium caseinate gels, but not that of heated milk gels, even though protein cross-linking was detected in heated milk. AbTyr did not cross-link proteins in any of the studied milk protein systems. TG was superior to TrTyr in gels prepared of heated milk. In acidified heated milk and sodium caseinate, TrTyr and TG treatment resulted in a decrease of the pore size. Scanning electron microscopy revealed more extensive particle interactions in the heated milk gels with TG than with TrTyr.     

Micromorphological studies of surface densified wood

Scots pine (Pinus sylvestris L.) wood was surface densified in its radial direction in an open press with one heated plate to obtain a higher density on the wood surface whilst retaining the overall thickness of the sample. This study investigated the effect of temperature (100, 150 and 200 °C) and press closing speed (5, 10 and 30 mm/min, giving closing times of 60, 30 and 10 s, respectively) on the micromorphology of the cell-wall, as well as changes occurring during set-recovery of the densified wood. The micromorphology was analysed using scanning electron microscopy (SEM) combined with a sample preparation technique based on ultraviolet-excimer laser ablation. Furthermore, the density profiles of the samples were measured. Low press temperature (100 °C) and short closing time (10 s) resulted in more deformation through the whole thickness, whilst increasing the temperature (150 and 200 °C) and prolonging the closing time (30 and 60 s) enabled more targeted deformation closer to the heated plate. The deformation occurred in the earlywood regions as curling and twisting of the radial cell-walls, however, no apparent cell-wall disruption or internal fracture was observed, even at low temperatures and fast press closing speed, nor after soaking and drying of the samples. In the SEM-analysis after soaking and drying, it was noticed that the cells did not completely recover their original form. Thus, part of the deformation was considered permanent perhaps due to viscoelastic flow and plastic deformation of the cell-wall components.