Co-planting can phytoextract similar amounts of cadmium and zinc to mono-cropping from contaminated soils

Co-planting crops normally decreases the main crop yield due to the reduced soil surface area occupied by the main crop. However, in our previous experiments, co-planting Sedum alfredii, a shade-requiring, Cd and Zn-hyperaccumulating plant, with corn increased the biomass and metal phytoextraction of S. alfredii. This experiment was conducted to verify if co-planting another hyperaccumulator, Thlaspi caerulescens, with ryegrass (Lolium perenne) in a pot-trial could obtain a similar result. The soil was separated by two permeable nets with a 2 mm interface soil layer to obtain a shared rhizosphere zone. Soluble metal concentrations in the soil in different rooting zones were measured using 0.01 mol L^?1 CaCl₂ extraction. The results showed that the growth of T. caerulescens was significantly promoted by co-planting, with a growth increase of about 2-fold compared with monoculture growth. The total uptake of Cd and Zn by T. caerulescens was not decreased by co-planting, and resulted in similar phytoextraction rates for Cd (about 26.6% of the soil total Cd) and Zn (about 2.4% of the soil total Zn) when compared with monoculture, though the T. caerulescens population was decreased by 50% because of co-planting. Analysis of soil samples showed that T. caerulescens substantially reduced the concentrations of 0.01 mol L^?1 CaCl₂ extractable Cd and Zn throughout the soil, even in the interface area and the ryegrass rooting area. The ryegrass roots did not mobilize more metals for the co-planted T. caerulescens. Based on these results, existing grass on contaminated land could be partly left while planting metal hyperaccumulators for phytoremediation in order to reduce runoff from the contaminated soil. However a field scale trial would be required for these results to be verified.     

Wool-degrading <Emphasis Type="Italic">Bacillus</Emphasis> isolates: extracellular protease production for microbial processing of fabrics

Wool is a natural animal fiber commonly used in fabrics, but requires physical and chemical processing treatment for such applications. With the aim of developing new woollen textile products using environmentally friendly treatments, proteolytic bacteria were isolated from raw wool samples of Merino sheep and screened for wool-degrading activity. Two isolates were identified as Bacillus megaterium L4 and Bacillus thuringiensis L11 by 16S rRNA gene sequence analysis. Both isolates grew on a minimal medium using wool-fiber or wool-fabric as sole carbon and nitrogen sources. Bacterial growth was correlated with extracellular protease activity, and maximal protease production was in early stationary phase. The exoprotease produced by L11 was found to be a thermo-tolerant metalloprotease stabilized by calcium or magnesium, and had optimum activity at pH 7.0 and temperature at 40°C. During bacterial growth the wool-fiber lost weight, but it did not show changes in diameter. When wool-fabric was used instead of wool-fiber weight loss and non-shrinking was found. These are encouraging results for textile processing that should be useful for development of new textile products by direct microbial processing. A potential alternative that could be suggested from our study would be to treat wool with wool-degrading microorganisms in order to develop environmentally friendly processes.     

Impact of Legume Flour Addition on Pasta Structure: Consequences on Its <Emphasis Type="Italic">In Vitro</Emphasis> Starch Digestibility

Pasta is popular for its ease of cooking and its low glycaemic index (GI). This interesting nutritional property can be attributed to its specific compact structure generally described as a protein network entrapping starch granules. Despite this low GI, pasta is poor in fibres and lack some essential amino acids. To enhance its nutritional composition, pasta can be fortified with non-traditional ingredients such as legume flours. The objective of this study was to investigate the impact of legume flour addition on pasta structure and the inherent consequences on the in vitro digestibility of starch. The addition of a high level (35%, w/w) of legume flour, especially split pea flour, induced some minor structural changes in pasta. The inclusion of fibres, the dilution of gluten proteins by albumins and globulins, and the larger amount of thin protein films (in split pea pasta) may have favoured higher susceptibility of starch to digestive enzymes. At the opposite, the presence of some partially gelatinised starch granules in the core of fortified pasta may have favoured the decrease in the in vitro starch digestibility. As a consequence, a high level of legume flour addition in pasta did not have any significant impact on its in vitro starch digestibility. A high level of split pea and faba bean flours can thus be added to pasta to increase its nutritional composition while keeping its low glycaemic index.     

Acute dyspnoea and single tracheal localisation of mantle cell lymphoma

We report a case of a 48-year-old Chinese female with end-stage renal disease and chronic anemia on hemodialysis. Clonazepam was prescribed for myoclonus disorder two weeks prior to her hospitalization. Subsequently, she was hospitalized for neutropenic fever with thrombocytopenia and worsening anemia. Bone marrow examination demonstrated a markedly hypocellular marrow (10-20% total cellularity). Clonazepam was discontinued, with gradual improvement of thrombocytopenia, and neutropenia in 1-2 weeks. To our knowledge, this is the first reported case of pancytopenia associated with clonazepam. We recommend patients taking clonazepam to be monitored with regular complete blood count to check for clinically significant pancytopenia or thrombocytopenia.     

Increase in cellular pool of low-molecular-weight iron during ethanol metabolism in rat hepatocyte cultures

Ethanol-induced lipid peroxidation was studied in primary rat hepatocyte cultures supplemented with ethanol at the concentration of 50 mM. Lipid peroxidation was assessed by two indices: (1) conjugated dienes by second-derivative UV spectroscopy in lipid extract of hepatocytes (intracellular content), and (2) free malondialdehyde (MDA) by HPLC-UV detection and quantitation for the incubation medium (extracellular content). In cultures supplemented with ethanol, free MDA increased significantly in culture media, whereas no elevation of conjugated diene level was observed in the corresponding hepatocytes. The cellular pool of low-mol-wt (LMW) iron was also evaluated in the hepatocytes using an electron spin resonance procedure. An early increase of intracellular LMW iron (≤1 hr) was observed in ethanol-supplemented cultures; it was inhibited by 4-methylpyrazole, an inhibitor of alcohol dehydrogenase, whereas α-tocopherol, which prevented lipid peroxidation, did not inhibit the increase of LMW iron. Therefore, the LMW iron elevation was the result of ethanol metabolism and was not secondarily induced by lipid hydroperoxides. Thus, ethanol caused lipid peroxidation in rat hepatocytes as shown by the increase of free MDA, although no conjugated diene elevation was detected. During ethanol metabolism, an increase in cellular LMW iron was observed that could enhance conjugated diene degradation.     

DNA integrity and transgene expression after passage through the NOGA needle catheter used for therapeutic myocardial angiogenesis

BACKGROUND: The NOGA (Biosense Webster, Markham, ON, Canada) injection catheter is an innovative navigational device that provides an ideal platform for intra-myocardial injection material. However, injection through a long (1.91 m), narrow (27G) nitinol needle could result in deterioration in the integrity and functionality of DNA. METHODS: To test this possibility, DNA in plasmid form (pcDNA3.1) containing the Lac Z transgene (250 micro l) was passed through the NOGA needle using a hand-held 1 cc syringe at a gentle hand injection pressure (43 +/- 3 PSI, 3.0 +/- 0.2 kg/cm(2)) or at maximal manual pressure (90 +/- 6 PSI, 6.3 +/- 0.4 kg/cm(2)), either once or 20 times. This DNA, compared to DNA not passed through the NOGA needle (control), was then used to transfect primary cultures of rat skin fibroblasts (FB) from Fisher 344 rats and the cells were subsequently stained for beta galactosidase (betagal). RESULTS: Transfection efficiency was significantly reduced by passing the DNA through the needle at both 43 +/- 3 PSI (78 +/- 4% of control, n = 10, P < 0.05 versus control) and 90 +/- 6 PSI (66 +/- 4 % of control, n = 10, P < 0.01 versus control, P < 0.02 versus 43 +/- 3 PSI). Passage of the DNA through the NOGA needle 20 times resulted in a transfection efficiency of only 5 +/- 1% of control (n = 20, P < 0.1 x 10(-11) versus control). Capillary Electrophoresis revealed that the reduction in transfection efficiency was due to a conformational change in the DNA from predominantly supercoiled to nicked and linearized DNA. Transfection efficiency as compared with control decreased as the concentration of the DNA solution which was passed through the needle was increased from 0.3 micro g/ micro l to 2.4 micro g/ micro l. Recovery experiments confirmed that the reduction in transfection efficiency was not due to loss of DNA by binding to the NOGA needle. CONCLUSION: These results suggest that DNA is susceptible to shear forces when injected through the NOGA needle even at nominal clinical injection pressures, suggesting that careful and controlled injections will be required to achieve optimal gene integrity and expression.     

Aphids at crossroads: when branch architecture alters aphid infestation patterns in the apple tree

Plant architecture highly constrains pest infestation but is rarely considered in studies on plant–insect interactions. We analysed the relationships between apple tree architectural traits manipulated by tree training and within-branch development of Dysaphis plantaginea (rosy apple aphid, RAA), a major apple pest, during its multiplication wingless phase in spring. We hypothesised that the degree of branching had an effect on RAA within-branch infestation. In an experimental apple orchard, the infestation by aphid wingless forms was surveyed in two consecutive spring seasons within branches manipulated to design contrasted architectures differing in shoot numbers, shoot density and branching orders. Whatever the branch management system, aphid infestation was higher on long versus short, fruiting versus vegetative, and growing versus non-growing shoots. Either less infested shoots or less severe infestation were observed in the most branched system. A pattern of within-branch short-distance infestation was confirmed. Moreover, the number of branching points between two shoots exerted a high constraint on this infestation pattern. Beside possible trophic effects due to plant growth patterns already documented in the literature, a high degree of branching is likely to be a key-architectural trait to constrain within-branch aphid infestation. This opens new perspectives on the manipulation of branch architecture as a mean giving partial control of pests towards sustainable fruit production.     

Applications of a new subspace clustering algorithm (COSA) in medical systems biology

A novel clustering approach named Clustering Objects on Subsets of Attributes (COSA) has been proposed (Friedman and Meulman, (2004). Clustering objects on subsets of attributes. J. R. Statist. Soc. B 66, 1–25.) for unsupervised analysis of complex data sets. We demonstrate its usefulness in medical systems biology studies. Examples of metabolomics analyses are described as well as the unsupervised clustering based on the study of disease pathology and intervention effects in rats and humans. In comparison to principal components analysis and hierarchical clustering based on Euclidean distance, COSA shows an enhanced capability to trace partial similarities in groups of objects enabling a new discovery approach in systems biology as well as offering a unique approach to reveal common denominators of complex multi-factorial diseases in animal and human studies. Doris Damian, Matej Orešič, and Elwin Verheij contributed equally to this work.     

ER–plasma membrane contact sites contribute to autophagosome biogenesis by regulation of local PI3P synthesis

The double‐membrane‐bound autophagosome is formed by the closure of a structure called the phagophore, origin of which is still unclear. The endoplasmic reticulum (ER) is clearly implicated in autophagosome biogenesis due to the presence of the omegasome subdomain positive for DFCP1, a phosphatidyl‐inositol‐3‐phosphate (PI3P) binding protein. Contribution of other membrane sources, like the plasma membrane (PM), is still difficult to integrate in a global picture. Here we show that ER–plasma membrane contact sites are mobilized for autophagosome biogenesis, by direct implication of the tethering extended synaptotagmins (E‐Syts) proteins. Imaging data revealed that early autophagic markers are recruited to E‐Syt‐containing domains during autophagy and that inhibition of E‐Syts expression leads to a reduction in autophagosome biogenesis. Furthermore, we demonstrate that E‐Syts are essential for autophagy‐associated PI3P synthesis at the cortical ER membrane via the recruitment of VMP1, the stabilizing ER partner of the PI3KC3 complex. These results highlight the contribution of ER–plasma membrane tethers to autophagosome biogenesis regulation and support the importance of membrane contact sites in autophagy.