Determination of total N-nitroso compounds by chemical denitrosation using CuCl

A method for the determination of total N-nitroso compounds (NOC) by chemical denitrosation and subsequent chemiluminescence detection of evolved NO is described. Denitrosation was accomplished with CuCl in HCl at 70 degrees C. The detection limit for N-nitrosoproline (NPRO) was 1 pmol. NO formation from NPRO was linear (R(2) = 0.999) from 4 pmol to 2 nmol. Among the possible interfering compounds tested, only S-nitroso compounds contribute any significant interference. This method had several advantages over other similar methods: (1) A commercially available one-piece reaction vessel and a NO analyzer with software were used. (2) NO release occurred rapidly and was easily measured and quantified. (3) Compared to HBr or HI, CuCl was more convenient to work with and safe. (4) CuCl was suitable for samples in aqueous and most organic solvents. The application of this method to food, personal care products, and human body fluids demonstrates its utility.     

Activity and process stability of purified green pepper (Capsicum annuum) pectin methylesterase

Pectin methylesterase (PME) from green bell peppers (Capsicum annuum) was extracted and purified by affinity chromatography on a CNBr-Sepharose-PMEI column. A single protein peak with pectin methylesterase activity was observed. For the pepper PME, a biochemical characterization in terms of molar mass (MM), isoelectric points (pI), and kinetic parameters for activity and thermostability was performed. The optimum pH for PME activity at 22 degrees C was 7.5, and its optimum temperature at neutral pH was between 52.5 and 55.0 degrees C. The purified pepper PME required the presence of 0.13 M NaCl for optimum activity. Isothermal inactivation of purified pepper PME in 20 mM Tris buffer (pH 7.5) could be described by a fractional conversion model for lower temperatures (55-57 degrees C) and a biphasic model for higher temperatures (58-70 degrees C). The enzyme showed a stable behavior toward high-pressure/temperature treatments.     

Chemical discrimination of arabica and robusta coffees by Fourier transform Raman spectroscopy

This article deals with the potential of Fourier transform (FT) Raman spectroscopy in discrimination of botanical species of green and roasted coffees. There are two species of commercial importance: Coffea arabica (arabica) and Coffea canephora (robusta). It is recognized that they differ in their lipid fraction, especially in the content of the diterpene kahweol, which is present at 0.1-0.3% dry matter basis in arabica beans and only in traces (<0.01%) in robusta. The visual examination of the Raman spectra of the lipid fraction extracted from arabica, robusta and liberica samples shows differences in the mid-wavenumbers region: arabica spectra have two characteristic scattering bands at 1567 and 1478 cm(-1). The spectrum of the pure kahweol shows the same bands. Principal component analysis is applied to the spectra and reveals clustering according to the coffee species. The first principal component (PC1) explains 93% of the spectral variation and corresponds to the kahweol concentration. Using the PC1 score plot, two groups of arabica can be distinguished as follows: one group with high kahweol content and another group with low kahweol content. The first group includes samples coming from Kenya and Jamaica; the second group includes samples from Australia. The main difference between these coffees is that those from Kenya and Jamaica are well-known for growing at a high altitude whereas those ones from Australia are grown at a low altitude. To our knowledge, the application of Raman spectroscopy has never been used in coffee analysis.     

Microbial community changes in heathland soil communities along a geographical gradient: interaction with climate change manipulations

Climate change constitutes a serious threat for European heathlands as unlike other sources of damage, such as over-grazing, local remediation is not a possibility. Within the large pan-European projects, CLIMOOR and VULCAN, the effect of periodic drought and increased temperature were investigated in four heathland ecosystems along a geographical and climatic gradient across Europe. Fluorogenically labelled substrates for four enzymes (glucosidase, sulphatase, phosphatase, leucine amino peptidase) were used to measure extra-cellular enzyme activity in soil samples from each of the CLIMOOR sites. Microbial extra-cellular enzyme production is linked to microbial activity as well as soil physico-chemical properties, making soil enzymes one of the more reactive components of terrestrial ecosystems and potentially excellent indicators of soil microbial functional status and diversity.Across all sites and over all the substrates, organic matter content was exponentially, inversely related to enzyme activity. Although the increase in temperature produced by the CLIMOOR roofs was small (on average 0.9 °C), this was sufficient to increase enzyme activity in all sites (on average by 45%). The increase was within the range of seasonal variability at each of the sites. The effect of drought on enzyme activity was more pronounced in the Northern European sites than the southern European, and most moisture limited, site. This suggests that the effect of temperature increases may be observed across all regions; however, the soils of northern Europe may be more sensitive to changes in rainfall patterns than more moisture limited Southern European soils.     

Agronomic performance of no-tillage relay intercropping with maize under smallholder conditions in Central Brazil

In the context of conservation agriculture on small scale farms of the Brazilian Cerrado, we hypothesized that planting a cover crop in relay with a commercial crop improves the efficiency of use of available natural resources, increasing biomass for use as fodder without reducing the grain yield of the main crop. The objective of this study was to measure the performance of two intercropped systems in terms of total above-ground biomass production and maize (Zea mays) grain yield: pigeon pea (Cajanus cajan) and Brachiaria (Brachiaria ruziziensis) sown as cover crops in established maize under a no-tillage management. The cover crops were sown at two different dates and a comparison was made with the three crops sown as a sole crop at the early sowing date. The experiment was conducted during the 2007-2008 and 2008-2009 growing seasons. Maize grain yield was not reduced by the presence of the relay cover crops in comparison with maize as the sole crop, even when the cover crop was sown soon after maize emergence. In contrast, the production of above-ground biomass by the cover crop was significantly lower when grown with maize than it was when grown as a sole crop. In the intercropped systems, when sown early, the cover crop produced higher total biomass than when sown late. Total above-ground biomass production of maize intercropped with a cover crop was much higher than that of any of the crops sown alone: the total biomass (average of the two growing seasons) produced by maize and pigeon pea was more than double that of maize grown alone. The land equivalent ratio (LER) of maize grain yield and biomass production was higher than one whatever the intercropped system used. It was particularly high when maize was intercropped with early sown pigeon pea; grain yield LER and biomass LER reaching, respectively, 1.72 and 1.73 in 2007-2008 and 2.02 and 2.03 in 2008-2009. These high LER values provide evidence for the complementary and the high efficiency of use of available resources by the intercropped plants and thus the advantage of such systems to produce both maize grain and cover crop forage under the conditions of our study.     

Morphinandienone alkaloids from Dehaasia longipedicellata

Five morphinoid alkaloids have been isolated from Dehaasia longipedicellata, namely (-) pallidine, a new alkaloid (+) pallidinine (1), (+)-milonine, (-) 8,14-dehydrosalutaridine and (-) sinoacutine.     

Infrared spectroscopy of comet 81P/Wild 2 samples returned by Stardust

Infrared spectra of material captured from comet 81P/Wild 2 by the Stardust spacecraft reveal indigenous aliphatic hydrocarbons similar to those in interplanetary dust particles thought to be derived from comets, but with longer chain lengths than those observed in the diffuse interstellar medium. Similarly, the Stardust samples contain abundant amorphous silicates in addition to crystalline silicates such as olivine and pyroxene. The presence of crystalline silicates in Wild 2 is consistent with mixing of solar system and interstellar matter. No hydrous silicates or carbonate minerals were detected, which suggests a lack of aqueous processing of Wild 2 dust.     

Diagnostic imaging for the assessment of acquired abdominal vascular diseases in small animals: A pictorial review

Advances in interventional radiology and surgical techniques now allow complex abdominal diseases to be more successfully treated in small animals. Abdominal vascular alterations, acquired as individual process or as complication of other lesions such as neoplasia, can be life‐threatening or at least greatly limit curative interventions of underlying diseases. Computed tomography (CT) and high‐definition ultrasonography are now readily available in veterinary referral centers. Yet, there is little information currently available on the use of these modalities for the diagnosis and characterization of these vascular alterations. The purpose of this article is to review the CT and ultrasonographic findings of acquired vascular diseases in the abdomen of dogs and cats, using both the veterinary and human medicine literature as references, and highlighting essential concepts through figures.     

Detection of Escherichia coli strains producing cytotoxic necrotizing factor type two (CNF2) by enzyme-linked immunosorbent assay

Sheep and rabbit antisera were produced against lysates of E. coli strain 711 (pVir). This K-12 strain carries the Vir plasmid which codes for Cytotoxic Necrotizing Factor type 2 (CNF2). Immunoglobulin G (IgG) fractions of both immune sera were subsequently purified by a two-step precipitation method. To increase the specificity for CNF2, the sheep IgG preparation was extensively adsorbed against both a sonicated extract of isogenic K-12 strain 711 and intact phenol-treated cells of vaccine strain 711 (pVir). An enzyme-linked immunosorbent assay (ELISA) was developed to detect clinical isolates of E. coli producing CNF2, using the final preparations of rabbit and sheep IgG in a double sandwich technique. The results obtained with this CNF2-ELISA were compared to those obtained with the conventional HeLa cell cytotoxicity assay. The testing of 133 E. coli strains (49 CNF2 positive strains and 84 negative strains) resulted in no false-negative and no false-positive. Therefore, the CNF2-ELISA offers a good alternative to the HeLa cell culture assay for the detection of CNF2-producing strains where facilities for and experience with cell cultures is lacking.