Effects of Brussels sprout juice on the cell cycle and adhesion of human colorectal carcinoma cells (HT29) in vitro

Consumption of Brassica vegetables is associated with a reduced risk of cancer of the alimentary tract in animal models and human populations. We used raw juice extracted from Brussels sprouts rich in the glucosinolate sinigrin to explore the effect of naturally occurring glucosinolate breakdown products on cell cycle progression and apoptosis in human colorectal carcinoma cells (HT29). Juice was prepared from sprout tissue immediately before use, and the glucosinolate breakdown products were determined by gas chromatography mass spectrometry and liquid chromatography mass spectrometry. The cell cycle was analyzed by flow cytometry on detached and adherent cells, and apoptosis was measured in the detached population by annexin V staining. Twenty-four hours after challenge with juice (10 microL/mL), 7-13% of adherent cells had detached from the substratum but the majority (82%) of these cells had not entered apoptosis, whereas only 33% of detached control cells were not apoptotic (p < 0.05). The main glucosinolate breakdown products were as follows: the sinigrin breakdown product, 1-cyano-2,3-epithiopropane (ca. 38 mM); the gluconapin hydrolysis product, 3-butenyl isothiocyanate (ca. 2.2.mM); the glucobrassicin metabolite, ascorbigen (ca. 8 mM); and low concentrations of other indole glucosinolate-derived hydrolysis products such as neoascorbigen and 3,3'-diindolylmethane. A variety of biologically active glucosinolate breakdown products are released by mechanical disruption of raw Brussels sprout tissue, but contrary to previous assumptions, allyl isothiocyanate is not the main compound responsible for the inhibition of cell proliferation.     

Bayesian estimation of cluster-level test accuracy based on different sampling schemes

We develop Bayesian models to estimate cluster-level test characteristics, sensitivity, specificity, prevalence, and predictive values, based on four different sampling schemes: a single test case and three sequential test cases. The corresponding cluster-level characteristics are calculated and compared for different sample sizes, sampling schemes, individual-level sensitivities, specificities, and cut-off values. We compared posterior estimates of individual-level and cluster-level characteristics for these four sampling schemes with simulated data. Two illustrations, one for Johne’s disease in cattle and another for Salmonella in pig herds, are used to demonstrate application of the methods.     

Characterisation of endobacterial communities in ectomycorrhizas by DNA- and RNA-based molecular methods

The diversity of endobacteria associated with ectomycorrhizas of Suillus variegatus and Tomentellopsis submollis, in two Corsican pine (Pinus nigra) stands was analysed by cultivation-dependent and cultivation-independent molecular methods. Denaturing gradient gel electrophoresis (DGGE) analysis revealed the cultivable endobacterial communities associated with S. variegatus were similar within the same stand. The most abundant cultivable bacterial species belonged to the genera Pseudomonas and Burkholderia. Cultivation-independent molecular analysis indicated that the structure of the endobacterial communities in ectomycorrhizas was consistent across all samples regardless of ECM fungal species or the pine stand from which the samples were collected. However, comparison between rDNA- and rRNA-derived DGGE gels showed that metabolically active endobacterial species were not always detected in rDNA-based profiles. Clone libraries constructed from rRNA molecules indicated that Pseudomonas and Burkholderia spp. were metabolically active bacteria. As some of the most abundant cultivable bacteria, including Bacillus/Paenibacillus spp., were not detected in cultivation-independent DGGE profiles, a combination of cultivation-dependent and -independent approaches provided a more complete assessment of the diversity of endobacteria associated with ectomycorrhizas.     

Ca2+ Effects on K+ Fluxes in Arabidopsis Seedlings Exposed to Al3+

Potassium transport was investigated in the root elongation zone of Arabidopsis seedlings during the first minutes of Al³⁺ exposure, using the non-invasive MIFE microelectrode technique. To prevent pH changes during Al³⁺ application, and to separate aluminium from acidic stress, plants were pre-treated with 5 mM homoPIPES before addition of AlCl₃ (pH 4.2). The 30-min treatment with 50 or 500 μM AlCl₃ led to a significant increase in K⁺ efflux in solutions containing 100 μM CaCl₂. This efflux was suppressed by high concentrations of Ca²⁺ (10 mM) in the bathing solution. Our results suggest that elevated external Ca²⁺ activities can sustain K⁺ influx in the root elongation zone during Al³⁺ exposure either by maintaining [Ca²⁺]_cyt or by affecting Al³⁺ uptake across the plasma membrane.     

Hydroxylysyl pyridinoline cross-link concentration affects the textural properties of fresh and smoked Atlantic salmon (Salmo salar L.) flesh

A simple HPLC method is presented to quantify the low concentration of hydroxylysyl pyridinoline (PYD) cross-links in Atlantic salmon (Salmo salar L.) muscle. The method involved the extraction of tissue with NaOH prior to hydrolysis, which greatly reduced the amount of protein to be hydrolyzed and made downstream operations easier and more reproducible. The concentration of PYD was 426 pmol g(-)(1) dry mass muscle in post-rigor muscle stored at 0 degrees C and sampled 3 d after death. Hydroxproline (HYP) concentration was determined following NaOH extraction as a measure of collagen content. In post-rigor samples, the alkaline-insoluble HYP fraction comprised 18.3% of the total HYP. Scanning electron microscopy revealed shrinkage of muscle fibers and a retraction of the connective tissue matrix in smoked salmon. PYD concentration was relatively resistant to processing to the smoked product, decreasing by around 11.7%, as compared to a 22.2% decrease in HYP. There was a positive correlation between PYD concentration and the firmness of post-rigor muscle samples as measured by an instrumental texture analyzer, explaining 25% of the total variation. A weaker but still significant correlation was found between PYD concentration and firmness in the smoked product. There was no relationship between fillet firmness and total collagen concentration, although the correlation with HYP in the alkaline-insoluble fraction was significant at the 6% level (P = 0.057). Our results indicate that only 1-3% of collagen molecules are linked by nonreducible mature cross-links in harvest size farmed Atlantic salmon and that PYD concentration is an important raw material characteristic for flesh quality.     

Note on the methods for determination of chromic oxide in shrimp feeds.

Chromic oxide is used as an inert marker to measure apparent digestibility of feeds in insects, terrestrial, and aquatic animals. Quantitative determination of chromic oxide content in the sample requires the oxidation of water insoluble trivalent chromic oxide to its water-soluble hexavalent form. The two commonly used oxidizing agents are 70% perchloric acid or a mixture of sodium molybdate, sulfuric, and perchloric acid. Chromic oxide content of the oxidized solution is then measured against known standards either directly by spectrophotometry in the visible range at 350, 370, or 440 nm or after forming a colored complex with diphenylcarbazide (DPC) by colorimetry at 540 nm. This study compared the two methods of oxidation followed by spectrophotometry at the three wavelengths and by DPC colorimetry. DPC colorimetry gave precise results than the direct measurement of dichromate ion, irrespective of the method of oxidation used. Ash from samples oxidized by perchloric acid and quantified by DPC colorimetry gave a better measure of actual chromic oxide content as compared to the other methods tested.     

Effect of spraying techniques on the brown rot of peach fruit,and on black spot,powdery mildew and the two-spotted mite of apple trees

Application by hand-held equipment of the commercially recommended doses of proprietary formulations of captan (9.0 g a. i. per tree), mancozeb (10.8 g a. i. per tree), oxythioquinox (1.1 g a. i. per tree), and cyhexatin (1.8 g a. i. per tree), in spray volumes ranging from 0.225 to 9.0 litres per tree, gave commercial control of the brown rot of peach fruit, and of black spot (apple scab), powdery mildew and the two-spotted mite (Tetranychus urticae) of apple trees. Commercial control was also obtained using a quarter of the recommended doses when applied in 0.225 or 2.25 litres per tree. The residue of oxythioquinox on the leaves, immediately after applying 1.1 g a. i. per tree, doubled as the volume of spray was reduced from 9.0 to 2.25 litres per tree, and increased further when the volume was reduced to 0.225 litres per tree. It is suggested that some of the oxythioquinox ran off the trees at the higher volumes, and that less run-off at lower volumes resulted in higher chemical residues.     

Novel chemical approaches to crop protection: Needs and solutions

The examination of conventional chemical methods of crop protection suggests problems and opportunities that call for alternative approaches. The need for such novel treatments is considered in relation to damaging organisms not controlled by traditional pesticides, biological availability, resistance, selectivity and compatibility with other methods within the concept of integrated pest, disease or weed management. Analysis of these needs and opportunities suggests directions in which to seek novel chemical agents including those which can attack vital processes not previously exploited as target sites, the modification of physicochemical properties determining mobility and availability, chemical formulation, symptom suppression and the manipulation of chemically-mediated processes other than direct toxic action, and the use of agents which extend the usefulness of existing toxic agents. Examples illustrating these approaches are discussed.