Evaluation of a commercial enzyme-based serum cholesterol test kit for analysis of phytosterol and phytostanol products

Plant sterols (phytosterols) have been shown to possess serum cholesterol-lowering properties. In recent years, several phytosterol-enriched functional food products have been developed and marketed. Some phytosterol products contain common unsaturated sterols and some contain a subset of phytosterols called phytostanols (saturated sterols, also called plant stanols). Current methods for the quantitative analysis of plant sterols are labor intensive and require sophisticated gas or liquid chromatographs. In this study, a popular commercial spectrophotometric serum cholesterol test kit was evaluated for the analysis of plant sterols. The results indicate that the method could be modified to analyze phytosterols and phytostanols by increasing the incubation time. Both free phytosterols and fatty acyl phytosteryl esters were quantitatively analyzed, but ferulate phytosteryl esters, such as those that are found in corn and other cereals, were not hydrolyzed by the enzymes in the test kit and therefore were not detected.     

Supplementation with grape seed polyphenols results in increased urinary excretion of 3-hydroxyphenylpropionic Acid, an important metabolite of proanthocyanidins in humans

Grape seed extract provides a concentrated source of polyphenols, most of which are proanthocyanidins. Polymeric proanthocyanidins are poorly absorbed in the small intestine of humans, and exposure may result from metabolism to phenolic acids by colonic bacteria. Any biological effects of proanthocyanidins may be due to the phenolic acid metabolites. Several phenolic acids have been identified as proanthocyanidin metabolites, but these may be derived from a range of other dietary sources. The aim of this study was to determine if 24-h urinary excretion of specific phenolic acids increased significantly and consistently following regular supplementation with grape seed extract. In a randomized, double-blind placebo-controlled trial, 69 volunteers received grape seed extract (1000 mg/day total polyphenols) or placebo for 6 weeks. Supplementation with grape seed polyphenols resulted in a consistent increase in the excretion of 3-hydroxyphenylpropionic acid (3-HPP, P < 0.001) and 4-O-methylgallic acid (P < 0.001) and a less consistent increase in the excretion of 3-hydroxyphenylacetic acid (P = 0.002). The observed increase in 3-HPP is in line with the suggestion that this compound is a major phenolic acid breakdown product of proanthocyanidin metabolism in vivo.     

Photooxidative degradation of beer bittering principles: a key step on the route to lightstruck flavor formation in beer

Isohumulones, dihydroisohumulones, tetrahydroisohumulones, and humulinones, important hop-derived bittering compounds in beer, were shown to give rise to reactive triacylmethyl radicals on interaction with triplet-excited riboflavin after spin trapping by 5,5-dimethyl-1-pyrroline N-oxide or 2-methyl-2-nitrosopropane, followed by electron paramagentic resonance spectroscopy combined with spectral simulation. Electron abstraction from the ionized beta-tricarbonyl chromophore, which is common to all five-membered ring hop derivatives, is the initial event on photoinduced degradation. Radicaloid decomposition of isohumulones leads to precursors for 3-methylbut-2-ene-1-thiol, the lightstruck constituent in beer. Interaction of reduced derivatives of isohumulones with triplet-excited riboflavin furnished radical precursors of volatile aldehydes, which may lead to the development of unpleasant stale or cardboard flavors.     

Micromorphology of a “welded” Sangamonian to Wisconsinan age paleosol in southwestern Wisconsin

Micromorphological characteristics indicate that in southwestern Wisconsin pedogenesis transgressed the Sangamonian-Wisconsinan chronostratigraphic boundary in conjunction with colluvial and eolian sedimentation. The use of micromorphology helps to distinguish between pedological, colluvial, and eolian components in the paleosol horizon sequence, which can be difficult to resolve and interpret solely from field investigations and from other laboratory analyses. Results show that a basal loess “mixed zone” within the welded paleosol profile contains features that are the products of colluvial reworking processes. We present micromorphology data which support stratigraphic relationships suggesting that colluvial processes were important in the formation of basal loess “mixed zones” in addition to other possible mixing processes such as pedoturbation and bioturbation.     

The origin and deposition history of polycyclic aromatic hydrocarbons in the finger lakes region of New York

Studies aimed at determining the deposition history of energy related, potentially carcinogenic, polycyclic aromatic hydrocarbons (PAHs) deposited in the sediments of Cayuga Lake, in the Finger Lakes Region of central New York State, are described. PAH fluxes in the 1850's were at least an order of magnitude less than the maximum rates in the period of 1940 to 1955. PAH fluxes began to decline before the Milliken coal fired power plant, located on Cayuga Lake, started service in 1955. Correlation between the PAH flux into the sediments and fossil fuel usage and emissions for the states of New York, Ohio and Pennsylvania and the United States as a whole, suggests that modern, high efficiency fossil fuel fired power plants are minor sources of PAHs compared to coal related residential heating and industrial combustion. Similar comparisons with historic emissions data do not support the hypothesis that PAH flux is a useful indicator of the combustion sources which produce the `acidifying' compounds SO₂ and NO_x for the Finger Lakes region.     

Hydrochemical modelling of a stream dominated by organic acids and organically bound aluminium

An existing hydrogeochemical model, the Birkenes model (BIM), has been extended to include production of dissolved organic carbon (DOC) in the soil/water system and its chemical interactions with H⁺ and A1³⁺. The model has been calibrated and verified using precipitation and runoff data from the Svartberget catchment in northern Sweden. The catchment is impacted by moderate amounts of sulphur deposition (6 kg S ha^?1 a^?1; the stream is high in DOC (7–35 mg L^?1) and experiences episodically low pH-values (～4.0). The refined BIM is able to simulate the main variations in major chemical components of the stream water (H⁺, Ca²⁺, Mg²⁺, Al_i, Al₀, Cl^?, SO₄ ^?2 and DOC), as well as stream discharge and ¹⁸O, while Na⁺ is not well reproduced. Although very simplified relationships are used for the chemical interactions between DOC and H⁺ and aluminium, the model shows that these interactions are essential to stream acidity and aluminiumspecies content. Some of the model parameters are poorly determined by calibration with runoff data only. Soil and soil water investigations may contribute to the evaluation and development of the model structure and the representation of chemical processes. Further improvements of the model should emphasize DOC-production/absorption, detailed studies on DOC chemical behaviour and hydrological structure.     

Population and behavioural level responses of arable soil earthworms to boardmill sludge application

The response of earthworms to soil application of boardmill waste sludge was quantified in field and laboratory experiments. The influence of one application of 6×10⁴ tonnes ha⁻¹ of unamended sludge was tested against no application on silty-clay arable soil. After 2 years, results in stubble-cultivated soil showed a 1.7 times lower density of earthworms where sludge was added, whereas there was no difference in average earthworm fresh weight. In direct-drilled soil, there was an indication of lower average numbers of Lumbricus terrestris middens when sludge was applied. Negative impacts in the field may have been due to indirect negative effects of the sludge application. In the laboratory, habitat choice tests were undertaken with two common species from the field site using sludge-based mulch and fertiliser products. Aporrectodea caliginosa showed no discernible preference between soil and soil mixed with unamended sludge (mulch), but favoured soil over soil mixed with sludge and chicken manure (fertiliser). Tests with L. terrestris showed a similar pattern. The preference of L. terrestris for the two products was compared with that for chopped barley straw by direct observation of foraging behaviour. Differences in foraging time of L. terrestris for different feeds were not significant, but mass of straw collected was significantly greater compared with either type of sludge treatment.     

The genetic diversity of Rhizobium leguminosarum bv. viciae in cultivated soils of the eastern Canadian prairie

Soil populations of Rhizobium leguminosarum bv. viciae (Rlv) that are infective and symbiotically effective on pea (Pisum sativum L.) have recently been shown to be quite widespread in agricultural soils of the eastern Canadian prairie. Here we report on studies carried out to assess the genetic diversity amongst these endemic Rlv strains and to attempt to determine if the endemic strains arose from previously used commercial rhizobial inoculants. Isolates of Rlv were collected from nodules of uninoculated pea plants from 20 sites across southern Manitoba and analyzed by plasmid profiling and PCR-RFLP of the 16S-23S rDNA internally transcribed spacer (ITS) region. Of 214 field isolates analyzed, 67 different plasmid profiles were identified, indicating a relatively high degree of variability among the isolates. Plasmid profiling of isolates from proximal nodules (near the base of the stem) and distal nodules (on lateral roots further from the root crown) from individual plants from one site suggested that the endemic strains were quite competitive relative to a commercial inoculant, occupying 78% of the proximal nodules and 96% of the distal nodules. PCR-RFLP of the 16S-23S rDNA ITS also suggested a relatively high degree of genetic variability among the field isolates. Analysis of the PCR-RFLP patterns of 15 selected isolates by UPGMA indicated two clusters of three field isolates each, with simple matching coefficients (SMCs) ≥0.95. However, to group all field isolates together, the SMC has to be reduced to 0.70. Regarding the origin of the endemic Rlv strains, there were few occurrences of the plasmid profiles of field isolates being identical to the profiles of inoculant Rlv strains commonly used in the region. Likewise, the plasmid profiles of isolates from nodules of wild Lathyrus plants located near some of the sites were all different from those of the field isolates. However, comparison of PCR-RFLP patterns suggested an influence of some inoculant strains on the chromosomal composition of some of the field isolates with SMCs of ≥0.92. Overall, plasmid profiles and PCR-RFLP patterns of the isolates from endemic Rlv populations from across southern Manitoba indicate a relatively high degree of genetic diversity among both plasmid and chromosomal components of endemic strains, but also suggest some influence of chromosomal information from previously used inoculant strains on the endemic soil strains.     

The restoration and re-creation of species-rich lowland grassland on land formerly managed for intensive agriculture in the UK

Intensive agriculture has resulted in the loss of biodiversity and the specialist flora and fauna associated with the semi-natural grasslands of low-intensity pastoral systems throughout northwest Europe. Techniques employed to restore and re-create these grasslands on agricultural land in the UK are reviewed. Extensive cutting and grazing management have been shown to diversify improved swards and facilitate re-colonisation on ex-arable soils, although rates of re-assembly of plant communities with affinity to existing semi-natural grasslands have generally been slow. On former agriculturally improved swards, nutrient depletion has accelerated this process, especially where “gaps” for establishment have been created. Similarly, on ex-arable soils “nutrient stripping” and sowing with diverse seed mixtures has led to the rapid development of species-rich swards. On free draining brown earths such an approach may be required to restore grassland communities where soil phosphorous concentrations exceed semi-natural levels by more than 10 mg/l (using Olsen's bicarbonate extractant). However, the appropriateness of this threshold for other soil types requires further sampling. Although restored grasslands are likely to contribute to national biodiversity targets success will ultimately depend on the reinstatement of the communities and ecological functions of semi-natural references. Although this is technically feasible for a few plant assemblages, less is known about the re-assembly of microbial and faunal communities, or the importance of trophic interactions during grassland succession. As a consequence, more research is required on the functional attributes of semi-natural grasslands, as well as the methods required to restore localised types, novel nutrient depletion techniques, the “phased” introduction of desirable but poor-performing species and the performance of different genotypes during grassland restoration.