Developmental regulation of aldoxime formation in seedlings and mature plants of Chinese cabbage (Brassica campestris ssp. pekinensis) and oilseed rape (Brassica napus): Glucosinolate and IAA biosynthetic enzymes

The first steps in the biosynthesis of glucosinolates and indole-3-acetic acid (IAA) in oilseed rape (Brassica napus L.) and Chinese cabbage (Brassica campestris ssp. pekinensis) involve the formation of aldoximes. In rape the formation of aldoximes from chain-extended amino acids, for aromatic and aliphatic glucosinolate biosynthesis, is catalysed by microsomal flavin-containing monooxygenases. The formation of indole-3-aldoxime from l-tryptophan, the potential precursor of both indole-3-acetic acid and indolyl-glucosinolates, is catalysed by several microsomal peroxidases. The biosynthesis of glucosinolates and indole-3-acetic acid was shown to be under developmental control in oilseed rape and Chinese cabbage. No monooxygenase activities were detected in cotyledons or old leaves of either species. The highest monooxygenase activities were found in young expanding leaves; as the leaves reached full expansion and matured the activities decreased rapidly. The indole-aldoxime-forming activity was found in all of the tissues analysed, but there was also a clear decrease in foliar activity with maturity in leaves of rape and Chinese cabbage. Partial characterisation of the Chinese cabbage monooxygenases showed that they have essentially identical properties to the previously characterised rape enzymes; they are not cytochrome P450-type enzymes, but resemble flavin-containing monooxygenases. No monooxygenase inhibitors were detected in microsomes prepared from either cotyledons or old leaves.Abbreviations DHMet dihomomethionine - FMO flavin-containing monooxygenase - HPhe homophenylalanine - IAA indole-3-acetic acid - l-Phe l-phenylalanine - l-Trp l-tryptophan - MO monooxygenase - IAALD indole-3-acetaldehyde - IAOX indole-3-aldoxime - THMet trihomomethionine     

Sprouty proteins are targeted to membrane ruffles upon growth factor receptor tyrosine kinase activation. Identification of a novel translocation domain

Sprouty (Spry) was first identified in a genetic screen in Drosophila to be an antagonist of fibroblast growth factor and epidermal growth factor (EGF) signaling, seemingly by inhibiting the Ras/MAP kinase pathway. Data base searches lead to the identification and cloning of, to date, four mammalian sprouty genes. The primary sequences of the mammalian sprouty gene products share a well conserved cysteine-rich C-terminal domain with the Drosophila protein. The N-terminal regions, however, do not exhibit significant homology. This study aimed at determining the disposition of Spry proteins in intact cells before and after stimulation of the EGF receptor tyrosine kinase. Full-length or deletion mutants of Spry, tagged at the N termini with the FLAG-epitope, were expressed in COS-1 cells by transient transfection and analyzed by immunofluorescence microscopy before and after EGF stimulation of the cells. In unstimulated cells, the Spry proteins were distributed throughout the cytosol except for human Sprouty2 (hSpry2), which, although generally located in the cytosol, co-localized with microtubules. In all cases, the Spry proteins underwent rapid translocation to membrane ruffles following EGF stimulation. The optimal translocation domain was identified by deletion and immunofluorescence analysis to be a highly conserved 105-amino acid domain in the C-terminal half of the hSpry2 protein. The translocation of this conserved domain, based on hSpry2 data, was independent of the activation of phosphatidylinositol-3 kinase.     

Malaria and urbanization in sub-Saharan Africa

There are already 40 cities in Africa with over 1 million inhabitants and the United Nations Environmental Programme estimates that by 2025 over 800 million people will live in urban areas. Recognizing that malaria control can improve the health of the vulnerable and remove a major obstacle to their economic development, the Malaria Knowledge Programme of the Liverpool School of Tropical Medicine and the Systemwide Initiative on Malaria and Agriculture convened a multi-sectoral technical consultation on urban malaria in Pretoria, South Africa from 2nd to 4th December, 2004. The aim of the meeting was to identify strategies for the assessment and control of urban malaria. This commentary reflects the discussions held during the meeting and aims to inform researchers and policy makers of the potential for containing and reversing the emerging problem of urban malaria.     

Hydrology is reflected in the functioning and community composition of methanotrophs in the littoral wetland of a boreal lake

In lake ecosystems a major proportion of methane (CH(4) ) emissions originate from the littoral zone, which can have a great spatial variability in hydrology, soil quality and vegetation. Hitherto, spatial heterogeneity and the effects it has on functioning and diversity of methanotrophs in littoral wetlands have been poorly understood. A diagnostic microarray based on the particulate methane monooxygenase gene coupled with geostatistics was used to analyse spatial patterns of methanotrophs in the littoral wetland of a eutrophic boreal lake (Lake Kev?t?n, Eastern Finland). The wetland had a hydrology gradient with a mean water table varying from -8 to -25 cm. The wettest area, comprising the highest CH(4) oxidation, had the highest abundance and species richness of methanotrophs. A high water table favoured the occurrence of type Ib methanotrophs, whereas types Ia and II were found under all moisture conditions. Thus the spatial heterogeneity in functioning and diversity of methanotrophs in littoral wetlands is highly dependent on the water table, which in turn varies spatially in relation to the geomorphology of the wetland. We suggest that changes in water levels resulting from regulation of lakes and/or global change will affect the abundance, activity and diversity of methanotrophs, and consequently CH(4) emissions from such systems.     

Left ventricular torsion, energetics, and diastolic function in normal human aging

This study determined, for the first time, whether the effects of normal aging on systolic and diastolic left ventricular function in subjects without cardiovascular disease are related to underlying energetic defects. Cardiac magnetic resonance imaging with tissue tagging and (31)P spectroscopy was used to determine global structure, function, myocardial strains, and the phosphocreatine-to-ATP ratio (PCr/ATP) in 49 healthy subjects aged 20-69 yr. The three major abnormalities that developed with increasing age were the early filling percentage (EFP, the left ventricular volume increase from end systole to mid-diastole divided by stroke volume × 100), which decreased with age, indicating impaired early diastolic filling (r = -0.72, P < 0.0001), the torsion-to-shortening ratio (TSR, measure of subepicardial torsion exerting mechanical advantage over subendocardial shortening), which increased with age indicating relative subendocardial dysfunction (r = 0.44, P < 0.02), and the PCr/ATP (decreased with increasing age, r = -0.52, P < 0.003). EFP and TSR were strongly correlated (r = -0.63, P < 0.0001), although they were not related to PCr/ATP [EFP vs. PCr/ATP: r = 0.34, not significant (NS) and TSR vs. PCr/ATP: r = -0.3, P = NS]. In normal aging, changes in EFP and TSR likely share the same pathophysiology, although it is unlikely that energetics have a major role in the functional effects of aging.     

Comparative effects of insecticides with different mechanisms of action on Chrysoperla externa (Neuroptera: Chrysopidae): Lethal,sublethal and dose–response effects

The comprehensive knowledge that the delayed systemic and reproduction side effects can be even more deleterious than acute toxicity, has caused a shift in focus toward sublethal effects assessment on physiology and behavior of beneficial insects. In this study, we assessed the risks posed by some insecticides with different mode of action through lethal and delayed systemic sublethal effects on the pupation, adult emergence, and reproduction of the chrysopid Chrysoperla externa (Hagen, 1861; Neuroptera: Chrysopidae), an important predator in pest biological control. The maximum field recommended dose (MFRD) and twice (2×MFRD) for chlorantraniliprole, tebufenozide, and pyriproxyfen were harmless to C. externa. In contrast, all the tested chitin synthesis inhibitors (CSIs) were highly detrimental to the predator, despite of their lack of acute lethal toxicity. Therefore, the safety assumed by using IGRs toward beneficial insects is not valid for chrysopids. Dose–response data showed that although all CSIs have a similar mechanism of action, the relative extent of toxicity may differ (novaluron > lufenuron > teflubenzuron). For CSIs, the delayed systemic effects became obvious at adult emergence, where the predicted no observable effect dose (NOED) was 1/2 048 of the MFRD for novaluron (0.085 ng/insect), and 1/256 of the MFRD for both lufenuron (0.25 ng/insect) and teflubenzuron (0.6 ng/insect). Finally, this work emphasized the significance of performing toxicity risk assessments with an adequate posttreatment period to avoid underestimating the toxicities of insecticides, as the acute lethal toxicity assays may not provide accurate information regarding the long‐range effects of hazardous compounds.     

Alterations in carbohydrate metabolism and its regulation in PPARalpha null mouse hearts

Although a shift from fatty acids (FAs) to carbohydrates (CHOs) is considered beneficial for the diseased heart, it is unclear why subjects with FA beta-oxidation defects are prone to cardiac decompensation under stress conditions. The present study investigated potential alterations in the myocardial utilization of CHOs for energy production and anaplerosis in 12-wk-old peroxisome proliferator-activating receptor-alpha (PPARalpha) null mice (a model of FA beta-oxidation defects). Carbon-13 methodology was used to assess substrate flux through energy-yielding pathways in hearts perfused ex vivo at two workloads with a physiological substrate mixture mimicking the fed state, and real-time RT-quantitative polymerase chain reaction was used to document the expression of selected metabolic genes. When compared with that from control C57BL/6 mice, isolated working hearts from PPARalpha null mice displayed an impaired capacity to withstand a rise in preload (mimicking an increased venous return as it occurs during exercise) as reflected by a 20% decline in the aortic flow rate. At the metabolic level, beyond the expected shift from FA (5-fold down) to CHO (1.5-fold up; P < 0.001) at both preloads, PPARalpha null hearts also displayed 1) a significantly greater contribution of exogenous lactate and glucose and/or glycogen (2-fold up) to endogenous pyruvate formation, whereas that of exogenous pyruvate remained unchanged and 2) marginal alterations in citric acid cycle-related parameters. The lactate production rate was the only measured parameter that was affected differently by preloads in control and PPARalpha null mouse hearts, suggesting a restricted reserve for the latter hearts to enhance glycolysis when the energy demand is increased. Alterations in the expression of some glycolysis-related genes suggest potential mechanisms involved in this defective CHO metabolism. Collectively, our data highlight the importance of metabolic alterations in CHO metabolism associated with FA oxidation defects as a factor that may predispose the heart to decompensation under stress conditions even in the fed state.