Introduction of sulfhydryl groups and disulfide linkage to mungbean 8Sα globulin and effects on physicochemical and functional properties

Sulfhydryl groups and disulfide bond were introduced in mungbean's major storage protein, 8Sα globulin, by protein engineering to improve structural stability and functional properties. Five modified proteins or mutants (F59C, I99C, A213C, one free sulfhydryl group; I99C/A213, one disulfide bridge; F59C/I99C/A213C, one free sulfhydryl group and one disulfide linkage) were expressed in Escherichia coli at a yield similar to that of the unmodified protein or wild type (WT) in soluble form (38%). The number of introduced groups in the mutants was confirmed by Ellman analysis. Mutant and WT proteins exhibited similar elution patterns on gel filtration indicating their trimeric native conformation. Mutants had 2 to 3.8 °C higher T_m values than WT and were digested by chymotrypsin at 52–58% in 60 min but exhibited different digestion patterns. All mutants showed greater hardness of heat-induced gels than WT, especially I99C/A213C and F59C/I99C/A213C. Results indicate the improved structural stability of the modified 8Sα globulin.     

Spatial and temporal distribution of singlet oxygen in Lake Superior

A multiyear field study was undertaken on Lake Superior to investigate singlet oxygen ((1)O(2)) photoproduction. Specifically, trends within the lake were examined, along with an assessment of whether correlations existed between chromophoric dissolved organic matter (CDOM) characteristics and (1)O(2) production rates and quantum yields. Quantum yield values were determined and used to estimate noontime surface (1)O(2) steady-state concentrations ([(1)O(2)](ss)). Samples were subdivided into three categories based on their absorbance properties (a300): riverine, river-impacted, or open lake sites. Using calculated surface [(1)O(2)](ss), photochemical half-lives under continuous summer sunlight were calculated for cimetidine, a pharmaceutical whose reaction with (1)O(2) has been established, to be on the order of hours, days, and a week for the riverine, river-impacted, and open lake waters, respectively. Of the CDOM properties investigated, it was found that dissolved organic carbon (DOC) and a300 were the best parameters for predicting production rates of [(1)O(2)](ss). For example, given the correlations found, one could predict [(1)O(2)](ss) within a factor of 4 using a300 alone. Changes in the quantum efficiency of (1)O(2) production upon dilution of river water samples with lake water samples demonstrated that the CDOM found in the open lake is not simply diluted riverine organic matter. The open lake pool was characterized by low absorption coefficient, low fluorescence, and low DOC, but more highly efficient (1)O(2) production and predominates the Lake Superior system spatially. This study establishes that parameters that reflect the quantity of CDOM (e.g., a300 and DOC) correlate with (1)O(2) production rates, while parameters that characterize the absorbance spectrum (e.g., spectral slope coefficient and E2:E3) correlate with (1)O(2) production quantum yields.     

Extended study on the influence of z-value(s) of single and multicomponent time-temperature integrators on the accuracy of quantitative thermal process assessment

The possibilities and limitations of single- and multicomponent time-temperature integrators (TTIs) for evaluating the impact of thermal processes on a target food attribute with a Ztarget value different from the zTTI value(s) of the TTI is far from sufficiently documented. In this study, several thousand time-temperature profiles were generated by heat transfer simulations based on a wide range of product and process thermal parameters and considering a Ztarget value of 10 degrees C and a reference temperature of 121.1 degrees C, both currently used to assess the safety of food sterilization processes. These simulations included 15 different Ztarget=10 degrees CF121.1 degrees C values in the range 3 to 60 min. The integration of the time-temperature profiles with ZTTI values of 5.5 to 20.5 degrees C in steps of 1 degrees C allowed generation of a large database containing for each combination of product and process parameters the correction factor to apply to the process value FmultiTTI, which was derived from a single- or multicomponent TTI, to obtain the target process value 10 degrees CF121.1 degrees C. The table and the graph results clearly demonstrated that multicomponent TTIs with z-values close to 10 degrees C can be used as an extremely efficient approach when a single-component TTI with a z-value of 10 degrees C is not available. In particular, a two-component TTI with z1 and z2 values respectively above and below the Ztarget value (10 degrees C in this study) would be the best option for the development of a TTI to assess the safety of sterilized foods. Whatever process and product parameters are used, such a TTI allows proper evaluation of the process value 10 degrees CF121.1 degrees C.     

New developments in speciality polymers: polymeric stabilizers

Many new speciality polymers have been developed in the last few years. In this paper polymeric stabilizers (antioxidants, flame retardants and ultraviolet stabilizers) will be discussed. Polymeric antioxidants of the hindered-phenol type, copolymers of 2,6-ditertiarybutyl-4-vinyl(or isopropenyl)phenol with styrene, methyl methacrylate, or more importantly butadiene or isoprene have been prepared; hydrogenation of the latter copolymers gave copolymers of the two polymerizable phenolic antioxidants with ethylene or ethylene/propylene. The polymeric antioxidants have been blended with diene polymers and selected polyolefins and have improved the long-term oxidative stability of these polymers. Polymeric flame retardants have been prepared by copolymerizing styrene and/or acrylonitrile with acrylates and methacrylates of aliphatic bromine-containing alcohols or bromine-containing phenols. Polymers with polymer-bound flame retardants have a higher limiting oxygen index compared with the original polymer. A new class of polymerizable ultraviolet stabilizers has also been developed; these stabilizers are styryl, α-methylstyryl, acryloyl and methacryloyl derivatives of 2(2-hydroxyphenyl)2H-benzotriazoles. These monomers have been copolymerized with styrene, acrylates and methacrylates. 2(2-Hydroxyphenyl)2H-benzotriazoles substituted in the 4 position of the benzotriazole ring with hydroxyl, acetoxy or carboxyl groups suitable for incorporation into polyesters, polycarbonates, polyamides and epoxy resins have also been synthesized. All 2(2-hydroxyphenyl)2H-benzotriazole ultraviolet absorbers and the polymers into which they are incorporated have high light absorbency with γ_max between 330 and 350 nm and extinction coefficients in some cases as high as 4.5 × 10⁴ 1 mol^?1 cm^?1.     

The esterified plasma fatty acid profile is altered in early HIV-1 infection

The online version of the original article can be found at     

Analysis of volatiles evolved during high-temperature treatment of thermally stable polymers. I. Nitrogen-containing acetylene-terminated resin

Cured samples of a nitrogen-containing acetylene-terminated resin, N,N′-(1,3-phenylene-dimethylidene)bis(3-ethynylaniline), have been heated at 10°C/min up to 900°C in a pyroprobe attached to a gas chromatograph/mass spectrometer (GC/MS). Analysis of the volatiles evolved during heating identified both gases and higher boiling compounds. The major higher boiling compounds are benzene, toluene, xylene, aniline, benzonitrile, m-methylaniline, and m-methylbenzonitrile; the gases include ammonia, methane, and traces of carbon dioxide. Correlations between sample temperature and the evolution of each of these compounds have been made. The onset of all volatile formation occurs between 450 and 500°C. The higher boiling volatiles peak, then end by approximately 700°C, while the gases peak then fall off but are still being evolved at 900°C. Average weight loss measurements of 13.6% at 700°C and 15.7% at 900°C agree with previously published thermogravimetric analysis (TGA) data. © 1993 John Wiley & Sons, Inc.     

Efficient Total Chemical Synthesis of 13C=18O Isotopomers of Human Insulin for Isotope‐Edited FTIR

Isotope‐edited two‐dimensional Fourier transform infrared spectroscopy (2 D FTIR) can potentially provide a unique probe of protein structure and dynamics. However, general methods for the site‐specific incorporation of stable ¹³C=¹⁸O labels into the polypeptide backbone of the protein molecule have not yet been established. Here we describe, as a prototype for the incorporation of specific arrays of isotope labels, the total chemical synthesis—via a key ester insulin intermediate—of 97 % enriched [(1‐¹³C=¹⁸O)Phe^B24] human insulin: stable‐isotope labeled at a single backbone amide carbonyl. The amino acid sequence as well as the positions of the disulfide bonds and the correctly folded structure were unambiguously confirmed by the X‐ray crystal structure of the synthetic protein molecule. In vitro assays of the isotope labeled [(1‐¹³C=¹⁸O)Phe^B24] human insulin showed that it had full insulin receptor binding activity. Linear and 2 D IR spectra revealed a distinct red‐shifted amide I carbonyl band peak at 1595 cm^?1 resulting from the (1‐¹³C=¹⁸O)Phe^B24 backbone label. This work illustrates the utility of chemical synthesis to enable the application of advanced physical methods for the elucidation of the molecular basis of protein function.     

Effect of CO2 Exposure on the Chemical Stability and Mechanical Properties of BaZrO3‐Ceramics

The reactivity of BaZrO₃ with CO₂ has been addressed as one of the major challenges with BaZrO₃‐based electrolytes in protonic ceramic fuel cells. Here, we present a study of the effect of CO₂ exposure on BaZrO₃‐materials at elevated temperatures. Dense BaZr_1?xY_xO_3?x/2 (x = 0, 0.05, 0.1, 0.2) and BaCe_0.2Zr_0.7Y_0.1O_2.95 ceramics were prepared by sintering of powder prepared by spray pyrolysis. The Vickers indentation method was used to determine the hardness and estimate the fracture toughness of pristine materials as well as the corresponding materials exposed to CO₂. Formation of BaCO₃ on the surface of exposed ceramics was confirmed by X‐ray diffraction and electron microcopy. The reaction resulted in formation of Ba‐deficient perovskite at the exposed surface. The reaction with CO₂ was most pronounced at 650°C compared to the other temperatures applied in the study. The reactivity was also shown to depend on the Y‐content and the grain size and was most pronounced for BaZr_0.9Y_0.1O_2.95. The reaction with CO₂ was observed to have a profound effect on the fracture toughness of the ceramics, demonstrating a depression of the mechanical stability of the materials. The results are discussed with respect to the chemical and mechanical stability of BaZrO₃ materials, with particular emphasis on the composition and grain size.     

Solvent and Extraction Conditions Control the Assayable Phenolic Content and Antioxidant Activities of Seeds of Black Beans,Canola and Millet

The effects of extraction solvent and conditions on the total phenolic content (TPC) and antioxidant activity of black beans, canola and foxtail millet were investigated. The antioxidant activity was assayed using 2,2‐diphenyl‐1‐picrylhydrazyl radical scavenging activity (DRSA) and oxygen radical absorbance capacity (ORAC). Four solvent systems, namely 70 % acetone, 80 % ethanol, 80 % methanol and a mixture of acetone/methanol/water (7:7:6, v/v/v) were used. The extraction methods adopted in this study included refluxing, homogenization, cold extraction and sonication. The TPC as measured using the Folin Ciocalteu's method were 12.35–28.39, 2.43–16.73, and 1.78–5.06 µmol catechin equivalents/g dry matter (dm) for canola, black beans and foxtail millet, respectively. Aqueous acetone afforded the highest TPC for black beans and canola. Within the same solvent system used, the TPC, DRSA and ORAC obtained from different extraction techniques differed for black beans, canola and foxtail millet. The results demonstrated that the solvent system as well as method influenced the extraction of phenolic compounds and their antioxidant activities, depending on the type of matrix in which phenolics were embedded.     

Response of winter wheat to N-fertiliser: Quantitative relations for components of growth

Measurements were made of yield of dry matter, plant-N content, and the distribution of mineral-N down the soil profile in 10 fertiliser-N experiments. In one of them detailed measurements were made throughout growth. Rate of N-uptake by the crop was unaffected by the amount of mineral-N in the upper 90 cm of soil when it was above about 30 kg N ha^–1. The %N in plants that received ample N-fertiliser declined with increase in plant mass according to a previously derived equation. During senescence there was an apparent loss of N from the crop.N-nutrition in the different experiments had little effect on the partition of assimilate between grain and straw. At harvest grain and straw weights were well related by a linear model which had the same gradient but different intercepts for each experiment. Grain %N was about four times greater than straw %N. Regression analysis supported the view that high evaporative conditions or temperatures during the growing period induced earlier harvest dates, less grain relative to straw, and a higher %N in the plant when ample N-fertiliser was applied but not when N-fertiliser was withheld.Other analyses indicated that cereal roots were generally unable to extract mineral nitrogen from the soil when the concentration was less than about 0.18 kg N ha^–1 cm^–1, that at low levels of N-nutrition the recovery of available inorganic-N from soil by the grain and straw was about 80%, and that the average mineralisation rates from early spring to shortly after harvest date varied between 0.22 and 0.88 kg N ha^–1 d^–1 from site to site.