Optimized LC/MS/MS analysis of morphine and codeine in poppy seed and evaluation of their fate during food processing as a basis for risk analysis

The opiate alkaloids present in poppy seed intended for use in food recently have raised major concerns. An efficient method for routine analysis of morphine and codeine using liquid chromatography in combination with tandem mass spectrometry on a triple quadrupole instrument (LC/MS/MS) was therefore developed. The optimal sample preparation was found to be cold extraction of 10 g of unground poppy seed with 30 mL of methanol containing 0.1% acetic acid for 60 min shaken at 250 rpm. The fate of morphine during food processing was also studied. All experiments led to a significant reduction of morphine and codeine. For poppy cake only 16-50% of the morphine was recovered, and in poppy buns at the highest temperature (220 degrees C) only 3% of the original morphine content was found. Ground poppy seed showed significantly lower recoveries than untreated seed. Morphine elimination during food processing has to be taken into account in the current discussion about its maximum limits in poppy seed.     

Expression of TOLL-like receptors (TLR) by bovine antigen-presenting cells-potential role in pathogen discrimination?

Invading pathogens are controlled by the innate and adaptive arms of the immune system. Adaptive immunity, mediated by B and T lymphocytes, recognises pathogens via high affinity receptors. However, the establishment of a primary adaptive immune response is not rapid enough to eradicate invading microorganisms as it involves cell proliferation, gene activation and protein synthesis. More rapid defence mechanisms are provided by innate immunity, which recognises invading pathogens by germ-line-encoded pattern recognition receptors. Recent evidence shows that this recognition can mainly be attributed to the family of TOLL-like receptors (TLR). Binding of pathogen-associated molecular patterns to TLR induces the production of reactive oxygen and nitrogen intermediates, pro-inflammatory cytokines, and up-regulates expression of co-stimulatory molecules, subsequently initiating the adaptive immunity. In this paper, we will discuss the current knowledge with regards to the TLR, and in particular the bovine family of TLR. In addition, we will show the expression of TLR mRNA in bovine antigen-presenting cell subsets, summarise the discovery and the critical roles of TLR2 in host defence against Mycobacteria, and provide evidence for a mycobacteria species-specific response of bovine macrophages.     

The application of sexual coral recruits for the sustainable management of ex situ populations in public aquariums to promote coral reef conservation — SECORE Project

• 1. SECORE (SExual COral REproduction) Project is an initiative of public aquariums and research institutions to produce and exchange sexual coral recruits for the sustainable management of ex situ populations. Here we present the results of the initial three years (2002–2004).
• 2. Primary polyps (n=501) of corals (Acropora tenuis, Agaricia humilis, Favia fragum) were transported from Rotterdam Zoo to Cologne, Burgers', Hagenbeck and London Zoos, where development of juveniles was monitored for 10 months. All polyps were produced at Rotterdam Zoo from laboratory colonies (A. humilis, F. fragum), and from larvae generated from field collected gametes at Akajima, Okinawa, Japan (A. tenuis). Additionally, planulae of A. tenuis (n=1440) were transported from Rotterdam Zoo to Burgers' Zoo and to London Zoo to obtain primary polyps.
• 3. Larval settlement (A. tenuis) was observed to be 3.00 ± 2.57% (mean ± SD; n=1480) in 2002 and 17.36 ± 6.01% (mean ± SD; n=1480) in 2003, significantly lower compared to settlement at Rotterdam Zoo (57.84 ± 11.01% in 2003; mean ± SD, n=1480). High post‐transport survival rates of 95.18 ± 4.86% (mean ± SD; n=501) were observed in primary polyps of all species.
• 4. Juvenile survival (t=10 months; A. tenuis: 18.4–86.2%; A. humilis: 0–19.7%; F. fragum: 13.3–72.7%) differed significantly between institutions. Mean colony sizes (measured 10 months after transportation) were, in all cases, similar or higher to those reported from literature.
• 5. The results demonstrate the potential of this method to serve as an economical and sustainable alternative to existing mostly exploitative techniques for aquarium stocking. The use of sexual recruits provides an effective and low cost alternative, which is, in principle, applicable to all coral species.
• 6. The project was extended from 9 to 28 institutions across Europe, the USA and Japan in 2004.

Copyright © 2006 John Wiley & Sons, Ltd.     

Estimating genetic effects in maternal and paternal half-sibs from tetraploid-diploid crosses in <Emphasis Type="Italic">Musa</Emphasis> spp.

Ploidy manipulations and interspecific crosses have allowed considerable genetic progress in Musa breeding, but estimation of genetic parameters for parental selection remains a major challenge. This study aimed to determine the components of genetic variance and the relative contribution of genetic effects to phenotypic variation of yield and phenological traits in secondary triploid hybrids from tetraploid–diploid crosses. The hybrids were evaluated in two consecutive growth cycles on a tropical forest site. Non-genetic effects accounted for a large fraction of the variation observed for most traits, except bunch weight. Partitioning of genetic variance into additive and dominance components confirmed the predominant role of additive genetic effects on the expression of bunch weight, fruit filling time, fruit length, plant height, and number of leaves while primarily non-additive effects accounted for suckering behavior and fruit circumference. Maternal general combining ability (GCA) effects accounted for the additive genetic variation in plant height and number of leaves, suggesting that selection for these traits should be carried out in tetraploid clones. Conversely, paternal GCA effects were the primary causes of genetic variation for fruit filling time, bunch weight, and fruit length, suggesting that these characters should be selected for in diploid clones. Specific combining ability (SCA) effects were observed for all traits, except fruit filling time, suggesting that additional genetic gain could be achieved through recombinative heterosis for these traits.     

Structural and physiological changes associated with the skin spot disorder in apple

Skin spot is an important physiological disorder of ‘Elstar’ apples (Malus × domestica Borkh.) that occurs after fruit have been removed from controlled atmosphere storage. Skin spots are irregular patches of small, round, brown blemishes. Cross-sections reveal a browning of protoplasts (coagulated) and of cell walls that extends into the hypodermis. Skin spots are always associated with linear, gaping and non-gaping microcracks in the cuticle. Staining of apple skin with calcofluor white usually results in white fluorescence of cell walls but, within a skin spot, the white fluorescence is weak or absent. Cell walls within, and in the immediate vicinity of skin spots stain with phloroglucin/HCl indicating the presence of lignin. The area of skin affected by skin spots was positively and linearly correlated with the area of the non-blush fruit surface infiltrated by acridine orange. In general, skin spots were limited to the non-blush fruit surface and occurred more frequently near the stem-end than the calyx region of the fruit. Skin spot areas were correlated with a 2.5-fold increase in water vapour permeability compared with unaffected areas (23.8 ± 4.0 m s⁻¹ with skin spots, 9.6 ± 2.1 × 10⁻⁵ m s⁻¹ without skin spots). Strips of the fruit skin from regions with skin spots had an increased maximum force and modulus of elasticity. Dipping fruit in ascorbic acid (0.1 or 0.3 mM for 10 min) before storage decreased the area affected by skin spots. There was no effect of dipping in ethanol/water (70%, v/v, 15 min) or in solutions of captan (1.5 g L⁻¹, 10 min) or trifloxystrobin (0.1 g L⁻¹, 10 min). In contrast, prestorage treatment with 1-methylcyclopropene (630 nL L⁻¹ for 24 h) or poststorage incubation in H₂O₂ (10% for 2, 6, 10 and 16 h) increased skin spots. Our data are consistent with a typical cell response to an oxidative burst that seems to be focussed on particular regions of the ‘Elstar’ fruit surface by concentrations of cuticular microcracks, and that is possibly caused by reoxygenation injury upon removal from CA storage.     

Identification of quantitative trait loci (QTLs) associated with maintenance of wheat (Triticum aestivum Desf.) quality characteristics under heat stress conditions

The goal of this study was to identify quantitative trait loci (QTLs) associated with the maintenance of wheat grain quality following post-anthesis heat stress in a recombinant inbred line (RIL) population. The response to heat stress was measured using the sodium dodecyl sulfate sedimentation test (SDSS), a significant predictor of bread baking quality. SDSS scores were used to identify QTLs associated with grain quality and QTLs associated with quality stability were identified based on percent change in SDSS score between the heat stress and control treatments. Four QTLs were identified, located one each on linkage groups 1B, 1D, 4A, and 7A. The 1B, 1D, and 4A QTLs were associated with grain quality; the QTL on linkage group 7A was associated with quality stability. To confirm the detected QTLs, eighty advanced lines grown at three Texas nurseries were tested for relationships between allelic polymorphism at QTL linked markers and quality traits. Quality trait stability in the advanced lines was estimated using the coefficient of variability (CV%) of quality traits between nurseries. The analysis supported the relationship of the predicted QTLs on linkage groups 1B, 1D, and 4A with quality traits. The confirmed QTLs may be used in marker assisted selection (MAS) to develop wheat lines possessing superior quality traits. In addition, identification of genetic regions associated with this trait will aid the identification of the underlying genes.     

Vp1 expression profiles during kernel development in six genotypes of wheat, triticale and rye

During the last few decades, the physiological and genetic background of dormancy, and correlated pre-harvest sprouting (PHS) have been intensively investigated. Special attention has often been paid to genetic factors that may explain and predict PHS susceptible behaviour. A major candidate is the Vp1 gene which is involved in embryo development and maturation as well as in dormancy establishment. In this study, Vp1 gene expression during kernel development was studied in wheat, triticale and rye as a potential biomarker for selecting PHS tolerant varieties in cereal breeding programs. Plants of known PHS tolerant and PHS susceptible varieties were grown under controlled conditions from flowering until harvest ripeness. During that period, kernels were regularly harvested for RNA extraction and cDNA synthesis. Calibrated and normalized relative Vp1 expression levels were obtained in an RT-qPCR assay. During kernel development, Vp1 expression levels generally showed a typical peak during the soft dough stage, after which they decreased and remained low until harvest maturity. Differences in Vp1 expression levels could be observed between the PHS susceptible and PHS tolerant varieties of wheat, with the PHS tolerant variety showing higher levels of relative Vp1 expression compared to the PHS susceptible variety. In triticale, however, this difference was only seen once and could not be confirmed in further experiments. It seems that the Vp1 gene in triticale behaves in a similar way as in rye, in which no specific trends could be observed.     

Comment on "Nonreciprocal light propagation in a silicon photonic circuit"

We show that the structure demonstrated by Feng et al. (Reports, 5 August 2011, p. 729) cannot enable optical isolation because it possesses a symmetric scattering matrix. Moreover, one cannot construct an optical isolator by incorporating this structure into any system as long as the system is linear and time-independent and is described by materials with a scalar dielectric function.     

Innovations in the diagnosis of infectious agents of veterinary importance--a seven-league boots fairy tale?

The development of infectiological diagnostic methodology is characterized by saltatoric, revolutionary leaps. Improved sensitivity and specificity, reduction of analytical sample volumes, increased sample throughput, reduced test complexity associated with improved user friendliness are hallmarks of such developments. Apart from purely technical amenities some innovations also enable conceptual adaptations of monitoring strategies. These include the use of diagnostic assays on-site, e. g. in the stable or in the practice. Selecting the assay which is most "fit-for-purpose" among the plethora of sometimes loudly advertised new technology may be difficult even for the well trained and experienced diagnostician. Standardized licensing procedures for infectiological diagnostic kits may be required to guarantee minimal norms of diagnostic quality. At the same time availability of a diversity of diagnostic kits is needed to broaden the base of diagnostic reliability. New infectiological diagnostic innovations will only be useful if the (expanded) results are transposable to the improved practical care of companion animals and farm animal holdings. In addition, the judgment of results, e. g. for epidemiological plausibility, remains a very important issue even when the test results have been produced by novel high technology diagnostics.