Polyphenol Oxidase in Wheat Grain: Whole Kernel and Bran Assays for Total and Soluble Activity

Color is a key quality trait of wheat products, and polyphenol oxidase (PPO) is implicated as playing a significant role in darkening and discoloration. In this study, total and soluble PPO activities were characterized in whole kernel assays and bran extracts. In whole kernel assays similar to AACC Approved Method 22–85, four wheat cultivars were ranked the same for both total and soluble (leached) PPO activity with L‐DOPA (diphenol) as the substrate. Total kernel PPO activity was much greater than soluble PPO activity in three hexaploid wheat cultivars, indicating that insoluble PPO was the major contributor to kernel PPO measurements. Tyrosine (monophenol) was an excellent PPO substrate in kernel assays as expected but had no activity as a substrate for soluble PPO. However, soluble PPO activity with tyrosine was activated by the addition of the diphenols chlorogenic acid and caffeic acid. When PPO was assayed in homogenized bran, 89–95% of total PPO activity remained insoluble, associated with the bran particles. The kernel assay detected <2% of PPO measured in an equivalent amount of homogenized bran. However, total PPO activity was 2‐fold higher in Klasic than in ID377s, both when measured in the kernel assay and in homogenized bran, indicating that the kernel assay was an accurate predictor of relative total extracted PPO activity in these two cultivars. Adding detergents (0.1% SDS plus 0.2% NP‐40) to the bran extraction buffer increased both soluble and insoluble PPO activity. Results indicate that relative PPO activities among wheat cultivars are similar in whole kernel and kernel leachate assays, and that the predominant insoluble fraction of PPO, which is relatively uncharacterized, may be largely responsible for wheat product discoloration.     

Characterization of a Unique “Super Soft” Kernel Trait in Wheat

Kernel texture in wheat (Triticum sp.) is central to end‐use quality and utilization. Here we report the discovery of a novel soft kernel trait in soft white winter wheat (T. aestivum L.). Two heritable kernel phenotypes were selected among F₃‐derived sibs, hereafter designated “normal soft” (wild‐type) and “super soft.” Normal soft lines exhibited single kernel characterization system (SKCS) hardness index (HI) values typical of soft wheat (HI ≈ 20), whereas the super soft lines were unusually soft (HI ≈ 5). Under some environments, individual super soft lines exhibited HI values as low as HI = –4. The super soft trait was manifested in reduced SKCS kernel texture and higher break flour yields, with some increase in sodium carbonate SRC (solvent retention capacity) values and sponge cake volumes. Straight‐grade flour yield, flour ash, milling score, and cookie diameter were largely unaffected. With the possible exception of the sodium carbonate SRC values, we observed no indication that the super soft trait conferred any negative aspects to commercial soft wheat quality. As such, the super soft trait may provide wheat breeders with new opportunities to modify the end‐use quality of wheat.     

Compressive Strength of Wheat Endosperm: Comparison of Endosperm Bricks to the Single Kernel Characterization System

The three major classes of endosperm texture (grain hardness) of soft and hard common, and durum wheat represent and define one of the leading determinants of the milling and end‐use quality of wheat. Although these three genetic classes are directly related to the Hardness locus and puroindoline gene function, much less is known about the kernel‐to‐kernel variation within pure varietal grain lots. Measurement of this variation is of considerable interest. The objective of this research was to compare kernel texture as determined by compression failure testing using endosperm bricks with results of whole‐kernel hardness obtained with the Single Kernel Characterization System 4100 hardness index (SKCS HI). In general terms, the variation obtained with the SKCS HI was of similar magnitude to that obtained using failure strain and failure energy of endosperm brick compression. Objective comparisons included frequency distribution plots, normalized frequency distribution plots, ANOVA model R², and coefficients of variation. Results indicated that compression testing and SKCS HI similarly captured the main features of texture classes but also reflected notable differences in texture properties among and within soft, hard, and durum classes. Neither brick compression testing nor the SKCS HI may be reasonably expected to correctly classify all individual kernels as to genetic texture class. However, modest improvements in correct classification rate or, more importantly, better classification related to end‐use quality may still be achievable.     

The effects of low pH,low calcium concentrations and elevated aluminium concentrations on sodium fluxes in brown trout,Salmo Trutta L.

The effects of pH (c. 7.0, 5.4, 4.5 and 4.0), nominal Al levels (0 and 8 μmol L^?1) and Ca levels (10 and 50 μmol L^?1) on Na influx, efflux and netflux of brown trout have been investigated using artificial lake water of known composition. Low pH had little effect on influx, but tended to increase efflux, particularly in the low Ca treatments. A nominal addition of 8 μmol Al L^?1 at pH 4.5 and 4.0 reduced influx significantly. Efflux was unaffected. Aluminium addition at pH c. 7.0 and 5.4 had no such effect. The measured Al concentrations at the end of the static 8 hr flux measuring experiments were markedly lower than the nominal amount of A1 added to the start.     

Effect of germination, seed abrasion and seed size on polyphenol oxidase assay activity in wheat

Polyphenol oxidase (PPO) is one of the major enzymes responsible for browning of wheat food products. Wheat cultivars differ in PPO activity and plant breeders wish to select germplasm and cultivars with low PPO activities. Prior to harvest, seeds may be exposed to conditions conducive to preharvest sprouting and mechanical damage may occur during harvesting and subsequent handling. Sprouting, mechanical damage and seed size may affect PPO assay activity and therefore impinge upon the breeder's ability to accurately select low PPO genotypes. The effects of incipient germination, mechanical abrasion and seed size on PPO assay activity were investigated using the cultivars ‘Klasic’(hard white spring, high PPO), ‘Penawawa’(soft white spring, high PPO) and ‘ID377s’(hard white spring, low PPO). The influence of embryo vs. distal portions of the seed on PPO assay activity was also examined. PPO activity was assayed with constant mixing using L‐3,4‐dihydroxyphenyl alanine (L‐DOPA) substrate at pH 6.5 at room temperature; absorbance of the reaction solution was measured at 475 nm. There was no change in PPO assay activity for seeds imbibed in water for up to 8 h. PPO assay activity increased in seeds imbibed from 8 to 16 h, and then gradually declined with increasing imbibing time. PPO assay activity initially increased as a result of mechanical abrasion but then gradually declined with increasing abrasion time. Large seeds had a higher total PPO assay activity than small seeds, but the difference in activity was not proportional to seed weight. There was only a slight difference in PPO assay activity between half‐seeds with or without the embryo. Overall, PPO assay activity in dry seeds was stable and exposure of seeds to injury and moisture did not make any significant change in PPO assay activity among the cultivars or their relative rankings.     

Nuclear Protective Action Advisories: A Policy Analysis and Evaluation

Widespread spontaneous evacuation, the tendency for people to evacuate even when not advised to do so, has been highlighted as one of the likely behavioral responses to a nuclear power plant emergency. Utility company representatives contend that protective action advisories can be structured so as to stifle the magnitude and geographic extent of spontaneous evacuation. Data from a utility-company-sponsored telephone survey of households on Long Island, New York, where the Shoreham Nuclear Power Station is located, are used in this paper to test this proposition. Analyses of responses given to three sets of increasingly serious reactor accident scenarios, with and without information instructing people what protective actions to take, raise serious questions about the ability of emergency notification messages to affect human behavior in a radiological emergency. The results suggest firstly, that even if people me specifically advised not to evacuate, most would be inclined to do so; and secondly, that attempts to stifle the propensity to evacuate among those who are not at risk are likely to increase the propensity to stay behind among those who are at risk and should evacuate.     

Detection of silica-rich deposits on Mars

Mineral deposits on the martian surface can elucidate ancient environmental conditions on the planet. Opaline silica deposits (as much as 91 weight percent SiO2) have been found in association with volcanic materials by the Mars rover Spirit. The deposits are present both as light-toned soils and as bedrock. We interpret these materials to have formed under hydrothermal conditions and therefore to be strong indicators of a former aqueous environment. This discovery is important for understanding the past habitability of Mars because hydrothermal environments on Earth support thriving microbial ecosystems.     

Crystal structure of a neutralizing human IGG against HIV-1: a template for vaccine design

We present the crystal structure at 2.7 angstrom resolution of the human antibody IgG1 b12. Antibody b12 recognizes the CD4-binding site of human immunodeficiency virus-1 (HIV-1) gp120 and is one of only two known antibodies against gp120 capable of broad and potent neutralization of primary HIV-1 isolates. A key feature of the antibody-combining site is the protruding, finger-like long CDR H3 that can penetrate the recessed CD4-binding site of gp120. A docking model of b12 and gp120 reveals severe structural constraints that explain the extraordinary challenge in eliciting effective neutralizing antibodies similar to b12. The structure, together with mutagenesis studies, provides a rationale for the extensive cross-reactivity of b12 and a valuable framework for the design of HIV-1 vaccines capable of eliciting b12-like activity.     

Role of chromosome 3A in stomatal resistance of winter wheat

Summary Leaf stomatal resistance, through transpiration and photosynthesis control, constitutes a major factor of productivity and adaptation in wheat. The aim of the investigations reported here was to identify chromosomal effects on the expression of the maximum stomatal resistance, determined under optimum conditions of irradiance and water supply. Leaf stomatal resistance was measured, on wheat grown in pots under natural and well-watered conditions, using a LI-COR LI-6200 portable photosynthesis system under a saturating light>1400 mol m^-1 s^-2. Reciprocal sets of chromosome substitution lines between two hard red winter wheat cultivars, Wichita and Cheyenne, were used to identify the chromosomes involved in the expression of this trait. The two parental cultivars were significantly different for the parameter investigated. Chromosome 3A appeared to be involved in the expression of the stomatal resistance value under optimum conditions. Taking into account the relationships previously established between this parameter and some agronomic traits, chromosome 3A might be involved directly in productivity-determining processes or in the adaptation to water conditions, in wheat.     

Insulin signaling in health and disease

The signaling pathways used by insulin have been identified [M. White, Insulin Signaling Pathway, Sci. STKE (Connections Map, as seen November 2003), http://stke.sciencemag.org/cgi/cm/cmp_12069]. Now our challenge is to understand how the failure of these signals is associated with obesity and the progressive failure of pancreatic beta cells that leads to diabetes. Whether better management of chronic inflammation can improve insulin action is an important area of investigation. Drugs that stimulate IRS2 (insulin receptor substrate protein 2) synthesis or signaling might be a good starting point. This knowledge will lead to rational strategies that prevent or cure diabetes.