Cell and nuclear degradation in sweetpotato [Ipomoea batatas (L.) Lam.] root meristems following exposure to salinity

Sodium chloride-induced cell and nuclear degradation in the root meristems of sweetpotato [Ipomoea batatas (L.) Lam.] were determined using fluorescent microscopy and flow cytometry analysis. Two sweetpotato cultivars were grown in liquid Murashige and Skoog medium and subjected to 0 mM and 500 mM NaCl, with or without 15 mM CaCl2, for periods up to 24 h. Changes to the nuclei of root meristematic cells showed a similar pattern of damage to the nuclei using both fluorescent microscopy and flow cytometry analysis. Damage occurring after only a few hours was followed by nuclear degradation at 24 h. Flow cytometry histograms showed a reduction in G1 and G2 nuclei and an increase in degraded nuclei in NaCl-stressed roots. Salinity-induced nuclear degradation was alleviated by the addition of CaCl2.

Optimisation of protoplast production in white lupin

The influence, was investigated, of abiotic parameters on the isolation of protoplasts from in vitro seedling cotyledons of white lupin. The protoplasts were found to be competent in withstanding a wide range of osmotic potentials of the enzyme medium, however, -2.25 MPa (0.5 M mannitol), resulted in the highest yield of protoplasts. The pH of the isolation medium also had a profound effect on protoplast production. Vacuum infiltration of the enzyme solution into the cotyledon tissue resulted in a progressive drop in the yield of protoplasts. The speed and duration of orbital agitation of the cotyledon tissue played a significant role in the release of protoplasts and a two step (stationary-gyratory) regime was found to be better than the gyratory-only system.

Effect of biotic factors on the isolation of Lupinus albus protoplasts

Several tissue types of Lupinus albus L. were investigated as sources for the isolation of protoplasts. Cotyledons from in vitro seedlings were found to yield the highest number of protoplasts compared with leaves, hypocotyls and roots. A combination of the protoplast isolation enzymes, cellulase and Pectolyase Y23, was capable of releasing the highest number of protoplasts compared with a combination of cellulase and Macerase. Protoplast yield increased with increasing cotyledon age but was accompanied by a progressive decline in protoplast viability. The optimal combination of protoplast yield and viability occurred when the protoplasts were isolated from 14- to 18-day-old cotyledons. The ratio between the volume of enzyme solution and the tissue biomass did not affect the protoplast production significantly. This is the first report of the isolation of protoplasts from a lupin cotyledon and, following the procedure described in this paper, an average yield of 1.2 x 10(6) protoplasts per gram of fresh tissue was obtainable.

Arthropod diversity and community structure in relation to land use in the mekong delta, vietnam

Declining biodiversity in agro-ecosystems, caused by intensification of production or expansion of monocultures, is associated with the emergence of agricultural pests. Understanding how land-use and management control crop-associated biodiversity is, therefore, one of the key steps towards the prediction and maintenance of natural pest-control. Here we report on relationships between land-use variables and arthropod community attributes (for example, species diversity, abundance and guild structure) across a diversification gradient in a rice-dominated landscape in the Mekong delta, Vietnam. We show that rice habitats contained the most diverse arthropod communities, compared with other uncultivated and cultivated land-use types. In addition, arthropod species density and Simpson's diversity in flower, vegetable and fruit habitats was positively related to rice cover in the local landscape. However, across the landscape as a whole, reduction in heterogeneity and the amount of uncultivated cover was associated, generally, with a loss of diversity. Furthermore, arthropod species density in tillering and flowering stages of rice was positively related to crop and vegetation richness, respectively, in the local landscape. Differential effects on feeding guilds were also observed in rice-associated communities with the proportional abundance of predators increasing and the proportional abundance of detritivores decreasing with increased landscape rice cover. Thus, we identify a range of rather complex, sometimes contradictory patterns concerning the impact of rice cover and landscape heterogeneity on arthropod community attributes. Importantly, we conclude that that land-use change associated with expansion of monoculture rice need not automatically impact diversity and functioning of the arthropod community.

In planta proteomics and proteogenomics of the biotrophic barley fungal pathogen blumeria graminis f. sp hordei

To further our understanding of powdery mildew biology during infection, we undertook a systematic shotgun proteomics analysis of the obligate biotroph Blumeria graminis f. sp. hordei at different stages of development in the host. Moreover we used a proteogenomics approach to feed information into the annotation of the newly sequenced genome. We analyzed and compared the proteomes from three stages of development representing different functions during the plant-dependent vegetative life cycle of this fungus. We identified 441 proteins in ungerminated spores, 775 proteins in epiphytic sporulating hyphae, and 47 proteins from haustoria inside barley leaf epidermal cells and used the data to aid annotation of the B. graminis f. sp. hordei genome. We also compared the differences in the protein complement of these key stages. Although confirming some of the previously reported findings and models derived from the analysis of transcriptome dynamics, our results also suggest that the intracellular haustoria are subject to stress possibly as a result of the plant defense strategy, including the production of reactive oxygen species. In addition, a number of small haustorial proteins with a predicted N-terminal signal peptide for secretion were identified in infected tissues: these represent candidate effector proteins that may play a role in controlling host metabolism and immunity. Molecular & Cellular Proteomics 8: 2368-2381, 2009.

Oestrogenic activity of benzyl salicylate, benzyl benzoate and butylphenylmethylpropional (Lilial) in MCF7 human breast cancer cells in vitro

Benzyl salicylate, benzyl benzoate and butylphenylmethylpropional (Lilial) are added to bodycare cosmetics used around the human breast. We report here that all three compounds possess oestrogenic activity in assays using the oestrogen-responsive MCF7 human breast cancer cell line. At 3 000 000-fold molar excess, they were able to partially displace [H-3]oestradiol from recombinant human oestrogen receptors ER alpha and ER beta, and from cytosolic ER of MCF7 cells. At concentrations in the range of 5 x 10(-5) to 5 x 10(-4) M, they were able to increase the expression of a stably integrated oestrogen-responsive reporter gene (ERE-CAT) and of the endogenous oestrogen-responsive pS2 gene in MCF7 cells, albeit to a lesser extent than with 10(-8) M 17 beta-oestradiol. They increased the proliferation of oestrogen-dependent MCF7 cells over 7 days, which could be inhibited by the antioestrogen fulvestrant, suggesting an ER-mediated mechanism. Although the extent of stimulation of proliferation over 7 days was lower with these compounds than with 10(-8) M 17 beta-oestradiol, given a longer time period of 35 days the extent of proliferation with 10(-4) M benzyl salicylate, benzyl benzoate or butylphenylmethylpropional increased to the same magnitude as observed with 10(-8) M 17 beta-oestradiol over 14 days. This demonstrates that benzyl salicylate, benzyl benzoate and butylphenylmethylpropional are further chemical components of cosmetic products which give oestrogenic responses in a human breast cancer cell line in culture. Further research is now needed to investigate whether oestrogenic responses are detectable using in vivo models and the extent to which these compounds might be absorbed through human skin and might enter human breast tissues. Copyright (C) 2009 John Wiley & Sons, Ltd.

HDC poinsettia tracker: flexible graphical tracking software

Growing pot poinsettia and similar crops involves careful crop monitoring and management to ensure that height specifications are met. Graphical tracking represents a target driven approach to decision support with simple interpretation. HDC (Horticultural Development Council) Poinsettia Tracker implements a graphical track based on the Generalised Logistic Curve, similar to that of other tracking packages. Any set of curve parameters can be used to track crop progress. However, graphical tracks must be expected to be site and cultivar specific. By providing a simple Curve fitting function, growers can easily develop their own site and variety specific ideal tracks based on past records with increasing quality as more seasons' data are added. (C) 2009 Elsevier B.V. All rights reserved.

Pliocene seasonality across the North Atlantic inferred from cheilostome bryozoans

Previous studies have shown an inverse correlation between zooid size in cheilostome bryozoans and ambient water temperature. This relationship underlies the MART technique which uses intracolonial variation in zooid size to predict mean annual range in temperature experienced by bryozoan colonies during their life. Here we apply the MART technique to study Early and Mid Pliocene bryozoans from Central America (Panama, Costa Rica), the USA (Florida, South Carolina, North Carolina, Virginia) and the UK (Suffolk) to reconstruct palaeoseasonality across a range of latitudes for the North Atlantic during the Pliocene Epoch. Compared to the present-day, our analyses suggest greater seasonality (ca 4.5 degrees C) in the southern Caribbean at the time of Cayo Agua Formation deposition (ca 4.25 Ma), in keeping with inferred upwelling prior to the closure of the isthmian barrier at 2.7 Ma. Bryozoans also indicate seasonal upwelling on the Gulf Coast of Florida in a similar manner to the present-day. Because upwelling can be highly localised and prone to spatial and temporal variation in the Gulf of Mexico today, it contributes little to a broad understanding of Pliocene North Atlantic waters. However, MART estimates for the coastal plain region indicate a general reduction in the annual range in temperature relative to the present, suggesting that the colder surface waters that today reach south to Cape Hatteras had less influence in Early to Mid Pliocene times. These results, along with evidence from other proxies, strongly support reduced seasonality and warmer conditions along the eastern seaboard of the USA in the Early to Mid Pliocene. Finally, the MART estimates amongst Coralline Crag localities provide evidence for an increased annual range in temperature in the southern North Sea than at present. Our study shows that bryozoan MART estimates provide a powerful, independent proxy for palaeoseasonality and is the first to demonstrate that the MART technique can be applied to infer palaeoclimates across a wide range of latitudes focusing on a variety of geological formations and geographical regions. Crown Copyright (C) 2009 Published by Elsevier B.V. All rights reserved.

Concentration of aluminium in breast cyst fluids collected from women affected by gross cystic breast disease

Gross cystic breast disease (GCBD) is the most common benign breast disorder, but the molecular basis of cyst formation remains to be identified. If the use of aluminium-based antiperspirant salts is involved in the etiology of gross breast cyst formation, it might be expected that aluminium would be at elevated levels in human breast cyst fluid (BCF). Aluminium was measured by ICP-MS in 48 samples of BCF, 30 samples of human blood serum and 45 samples of human breast milk at different stages of lactation (colostrum, intermediate, mature). The median level of aluminium in apocrine type I BCF (n:= 27, 150 mu g I-1) was significantly higher than in transuclative type II BCF (n = 21, 32 mu g I-1; P < 0.0001). By comparison, aluminium measurements gave a median concentration of 6 mu g I-1 in human serum and 25 mu g I-1 in human breast milk, with no difference between colostrum, intermediate and mature milk. Levels of aluminium were significantly higher in both types of BCF than in human serum (P < 0.0001). However when compared with human breast milk, aluminium levels were only significantly higher in apocrine type I BCF (P < 0.0001) and not in transudative type II BCF (P = 0.152). It remains to be identified why such high levels of aluminium were found in the apocrine type I BCF and from where the aluminium originated. However, if aluminium-based antiperspirants are found to be the source and to play any causal role in development of breast cysts, then it might become possible to prevent this common breast disorder. Copyright (C) 2008 John Wiley & Sons, Ltd.

Low molecular weight heparin significantly reduces embolisation after carotid endarterectomy - a randomised controller trial

Objectives The administration of unfractionated heparin (UFH) prior to carotid clamping during carotid endarterectomy (CEA) transiently increases the platelet aggregation response to arachidonic acid (AA) despite the use of aspirin. We hypothesized that this phenomenon might be reduced by using low molecular weight heparin (LMWH) resulting in fewer emboli in the early post-operative period. Methods 183 aspirinated patients undergoing CEA were randomised to 5000 IU UFH (n = 91) or 2500 IU LMWH (dalteparin, n = 92) prior to carotid clamping. End-points were: transcranial Doppler (TCD) measurement of embolisation, effect on bleeding and platelet aggregation to AA and adenosine 5′-diphosphate (ADP). Results Patients randomised to UFH had twice the odds of experiencing a higher number of emboli in the first 3 h after CEA, than those randomised to LMWH (p = 0.04). This was not associated with increased bleeding (mean time from flow restoration to operation end: 23 min (UFH) vs. 24 min (LMWH), p = 0.18). Platelet aggregation to AA increased significantly following heparinisation, but was unaffected by heparin type (p = 0.90). The platelets of patients randomised to LMWH exhibited significantly lower aggregation to ADP compared to UFH (p < 0.0001). Conclusions Intravenous LMWH is associated with a significant reduction in post-operative embolisation without increased bleeding. The higher rate of embolisation seen with UFH may be mediated by increased platelet aggregation to ADP, rather than to AA.

Use of global gene expression patterns in mechanistic studies of oestrogen action in MCF7 human breast cancer cells

Over the years, the MCF7 human breast cancer cell line has provided a model system for the study of cellular and molecular mechanisms in oestrogen regulation of cell proliferation and in progression to oestrogen and antioestrogen independent growth. Global gene expression profiling has shown that oestrogen action in MCF7 cells involves the coordinated regulation of hundreds of genes across a wide range of functional groupings and that more genes are down regulated than upregulated. Adaptation to long-term oestrogen deprivation, which results in loss of oestrogen-responsive growth, involves alterations to gene patterns not only at early time points (0-4 weeks) but continuing through to later times (20-55 weeks), and even involves alterations to patterns of oestrogen-regulated gene expression. Only 48% of the genes which were regulated >= 2-fold by oestradiol in oestrogen-responsive cells retained this responsiveness after long-term oestrogen deprivation but other genes developed de novo oestrogen regulation. Long-term exposure to fulvestrant, which resulted in loss of growth inhibition by the antioestrogen, resulted in some very large fold changes in gene expression up to 10,000-fold. Comparison of gene profiles produced by environmental chemicals with oestrogenic properties showed that each ligand gave its own unique expression profile which suggests that environmental oestrogens entering the human breast may give rise to a more complex web of interference in cell function than simply mimicking oestrogen action at inappropriate times. (C) 2009 Elsevier Ltd. All rights reserved.

A breakthrough in lupin biotechnology: prolific protocolonisation in recalcitrant white lupin (Lupinus albus) triggered by bovine serum albumin

Six nutrient formulations were studied for their efficacy in inducing mitosis in white lupin seedling cotyledon protoplasts of which the formulations of Schafer-Menuhr & Sturmer (AS) and Kao (K8p) were found to be superior over the other four when supplemented with 6-benzylaminopurine and alpha-naphthaleneacetic acid (alpha-NAA). An unltrafiltration treatment of K8p increased mitotic frequency by 130% when compared with the untreated control. Medium enrichment with 0.2% bovine serum albumin (BSA) brought about a dramatic 1341% rise in protoplast division in comparison with BSA-free medium but only when the enrichment was carried out in Kao and Michayluk (KM8p) background containing 2, 4-dichlorophenoxyacetic acid, alpha-NAA and zeatin. A higher number of protocolonies (each proliferating from single protoplast following multiple divisions) were seen in 0.4% BSA. With this breakthrough in white lupin protoplast research, it is now possible to reproducibly obtain protocolonies that was hitherto not possible.

Paraben esters: review of recent studies of endocrine toxicity, absorption, esterase and human exposure, and discussion of potential human health risks

This toxicology update reviews research over the past four years since publication in 2004 of the first measurement of intact esters of p-hydroxybenzoic acid (parabens) in human breast cancer tissues, and the suggestion that their presence in the human body might originate from topical application of bodycare cosmetics. The presence of intact paraben esters in human body tissues has now been confirmed by independent measurements in human urine, and the ability of parabens to penetrate human skin intact without breakdown by esterases and to be absorbed systemically has been demonstrated through studies not only in vitro but also in vivo using healthy human subjects. Using a wide variety of assay systems in vitro and in vivo, the oestrogen agonist properties of parabens together with their common metabolite (p-hydroxybenzoic acid) have been extensively documented, and, in addition, the parabens have now also been shown to possess androgen antagonist activity, to act as inhibitors of sulfotransferase enzymes and to possess genotoxic activity. With the continued use of parabens in the majority of bodycare cosmetics, there is a need to carry out detailed evaluation of the potential for parabens, together with other oestrogenic and genotoxic co-formulants of bodycare cosmetics, to increase female breast cancer incidence, to interfere with male reproductive functions and to influence development of malignant melanoma which has also recently been shown to be influenced by oestrogenic stimulation. Copyright (C) 2008 John Wiley & Sons, Ltd.