Comet assay application in environmental monitoring: DNA damage in human leukocytes and plant cells in comparison with bacterial and yeast tests.
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Urban airborne particulate is a complex mixture of air pollutants, many of which have not been identified. However, short-term mutagenesis tests together with chemicophysical parameter analysis are able to better assess air quality and genotoxic load. The findings of continuous monitoring (January 1991-August 1998) of urban air genotoxicity of a Po Valley town (Italy) on Salmonella typhimurium and Saccharomyces cerevisiae are reported. During this period, various measures (catalytic devices, unleaded fuels, annual vehicle overhaul, etc.) to improve air-dispersed pollutant control were enforced. However, a continuous presence of genotoxic compounds is shown and more qualitative than quantitative changes are evident. We also demonstrate the ability of the Comet assay to detect DNA-damaging agents in airborne particulate samples. We applied the test to human leukocytes and, with major improvements, to plant cells (Allium cepa roots and epigean tissues of Impatiens balsamina). The first findings on human leukocytes confirm the sensitivity of this assay, its peculiarity and its applicability in assessing genotoxicity in environmental samples. The capability of plants to show the response of multicellular organisms to environmental pollutants largely counterbalances a probable lowering in sensitivity. Moreover, application of the Comet test to epigean tissues could be useful in estimating the bioavailability of and genotoxic damage by air pollutants, including volatile compounds (ozone, benzene, nitrogen oxides, etc.) to higher plants. (+info)
Polyphosphates in intraradical and extraradical hyphae of an arbuscular mycorrhizal fungus, Gigaspora margarita.
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The amount of polyphosphate in the intraradical and extraradical hyphae of Gigaspora margarita was estimated from successive extractions with trichloroacetic acid (TCA), EDTA, and phenol-chloroform (PC). In the intraradical hyphae, most of the polyphosphate was present in TCA- and EDTA-soluble (short-chain and long-chain) fractions, whereas most of the polyphosphate in the extraradical hyphae was present in EDTA- and PC-soluble (long-chain and granular) fractions. (+info)
One of the origins of plasma membrane phosphatidylserine in plant cells is a local synthesis by a serine exchange activity.
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In plant cells, as in animal cells, the endoplasmic reticulum (ER) is considered to be the major site of phospholipid synthesis, and it has been shown that phosphatidylserine (PS) reaches the plasma membrane via the vesicular ER-Golgi-plasma membrane pathway in leek cells. However, it has never been determined whether the plasma membrane of leek cells is able to synthesize PS. We have analyzed the distribution of PS synthesizing enzymes along the vesicular pathway. In ER, Golgi and plasma membrane fractions isolated from leek cells, we have measured the activity of the two biosynthetic pathways leading to the synthesis of PS, i.e. serine exchange and CTP cytidylyltransferase plus PS synthase. We have found a high serine exchange activity in the plasma membrane fraction, and then determined that this membrane is able to synthesize both long chain fatty acid- and very long chain fatty acid-containing PS. Therefore, the PS in the plasma membrane of leek cells has two different origins: the intracellular vesicular pathway from the ER and a local synthesis in the plasma membrane. (+info)
Chronic consumption of raw but not boiled Welsh onion juice inhibits rat platelet function.
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Welsh onion has been consumed for prevention of cardiovascular disorders. To study if it has antithrombotic effects, 9-wk-old male Sprague-Dawley rats were studied. Some rats were fed raw or boiled Welsh onion juice (2 g. kg(-1). d(-1)) for 4 wk, and the remaining acted as the control. Before and after feeding, their systolic blood pressure was measured by a tail-cuff method. Two days after the treatment period, tail bleeding time, platelet function (including platelet aggregation and adhesion), plasma levels of prostaglandins, and platelet cyclic nucleotide levels were determined. In comparison to the control, raw Welsh onion juice consumption significantly (1) lowered resting systolic blood pressure; (2) prolonged the bleeding time; (3) diminished platelet adhesion on a fibrinogen-coated surface, ADP-evoked platelet aggregation and ADP-stimulated thromboxane release; (4) elevated the concentration of cyclic AMP, but not cyclic GMP, in platelets; (5) increased the plasma level of 6-keto-prostaglandin F(1alpha), the stable prostacyclin metabolite, but not the plasma nitrite level. On the contrary, boiled Welsh onion juice consumption was totally ineffective. In conclusion, consuming raw Welsh onion juice, but not boiled juice, has blood pressure lowering and antithrombotic effects in rats. These effects may be mediated by PGI(2)-cAMP pathway. (+info)
A novel alliinase from onion roots. Biochemical characterization and cDNA cloning.
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We have purified a novel alliinase (EC 4.4.1.4) from roots of onion (Allium cepa L.). Two isoforms with alliinase activity (I and II) were separated by concanavalin A-Sepharose and had molecular masses of 52.7 (I) and 50.5 (II) kD on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and 51 (I) and 57.5 (II) kD by gel filtration fast-protein liquid chromatography. Isoform I had an isoelectric point of 9.3, while isoform II had isoelectric points of 7.6, 7.9, 8.1, and 8.3. The isoforms differed in their glycosylation. Both contained xylose/fucose containing complex-type N-linked glycans, and isoform II also contained terminal mannose structures. Both isoforms had activity with S-alk(en)yl-L-cysteine sulfoxides. Unlike other allium alliinases, A. cepa root isoforms had cystine lyase activity. We cloned a gene from A. cepa root cDNA and show that it codes for A. cepa root alliinase protein. Homology to other reported allium alliinase genes is 50%. The gene coded for a protein of mass 51.2 kD, with two regions of deduced amino acid sequence identical to a 25- and a 40-amino acid region, as determined experimentally. The A. cepa root alliinase cDNA was expressed mainly in A. cepa roots. The structure and function of the alliinase gene family is discussed. (+info)
Dose estimations of fast neutrons from a nuclear reactor by micronuclear yields in onion seedlings.
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Irradiations of onion seedlings with fission neutrons from bare, Pb-moderated, and Fe-moderated 252Cf sources induced micronuclei in the root-tip cells at similar rates. The rate per cGy averaged for the three sources, , was 19 times higher than rate induced by 60Co gamma-rays. When neutron doses, Dn, were estimated from frequencies of micronuclei induced in onion seedlings after exposure to neutron-gamma mixed radiation from a 1 W nuclear reactor, using the reciprocal of as conversion factor, resulting Dn values agreed within 10% with doses measured with paired ionizing chambers. This excellent agreement was achieved by the high sensitivity of the onion system to fast neutrons relative to gamma-rays and the high contribution of fast neutrons to the total dose of mixed radiation in the reactor's field. (+info)
Radial hydraulic conductivity along developing onion roots.
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Although most studies have shown that water uptake varies along the length of a developing root, there is no consistent correlation of this pattern with root anatomy. In the present study, water movement into three zones of onion roots was measured by a series of mini-potometers. Uptake was least in the youngest zone (mean hydraulic conductivity, Lpr = 1.5 x 10(-7) +/- 0.34 x 10(-7) m MPa-1 s-1; +/- SE, n = 10 roots) in which the endodermis had developed only Casparian bands and the exodermis was immature. Uptake was significantly greater in the middle zone (Lpr = 2.4 x 10(-7) +/- 0.43 x 10(-7) m MPa-1 s-1; +/- SE, n = 10 roots) which had a mature exodermis with both Casparian bands and suberin lamellae, and continued at this level in the oldest zone in which the endodermis had also developed suberin lamellae (Lpr = 2.8 x 10(-7) +/- 0.30 x 10(-7) m MPa-1 s-1; +/- SE, n = 10 roots). Measurements of the hydraulic conductivities of individual cells (Lp) in the outer cortex using a cell pressure probe indicated that this parameter was uniform in all three zones tested (Lp = 1.3 x 10(-6) +/- 0.01 x 10(-6) m MPa-1 s-1; +/- SE, n = 60 cells). Lp of the youngest zone was lowered by mercuric chloride treatment, indicating the involvement of mercury-sensitive water channels (aquaporins). Water flow in the older two root zones measured by mini-potometers was also inhibited by mercuric chloride, despite the demonstrated impermeability of their exodermal layers to this substance. Thus, water channels in the epidermis and/or exodermis of the older regions were especially significant for water flow. The results of this and previous studies are discussed in terms of two models. The first, which describes maize root with an immature exodermis, is the 'uniform resistance model' where hydraulic resistances are evenly distributed across the root cylinder. The second, which describes the onion root with a mature exodermis, is the 'non-uniform resistance model' where resistances can be variable and are concentrated in a certain layer(s) on the radial path. (+info)
Effects on the growth of carrots (Daucus carota L.), cabbage (Brassica oleracea var. capitata L.) and onion (Allium cepa L.) of restricting the ability of the plants to intercept resources.
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The objective of this paper is to assess the size and penetration of edge effects in carrot, cabbage and onion field crops and the extent to which these edge effects are modified by the presence of aerial or soil competition between the crop rows. In all three crops, large weight differences developed between the plants in the edge rows and those in the central rows. There was no indication of plant weight fluctuating between large and small values with each successive row in from the edge, as suggested by others. In carrot and onion, edge effects were greatly reduced by the presence of either white reflective aerial partitions or soil partitions, indicating that these species competed for both light and soil resources in UK field conditions. In cabbage, the mere presence of clear aerial partitions between rows reduced edge effects and there was little effect of soil partitions. This indicates the predominance of shoot over root competition in this species. The differences between species are possibly related to the architectural flexibility of their shoots. These results suggest that, within crops, carrot and onion plants compete for light over a distance of about 20 cm in each direction and for below-ground resources over a distance of about 50 cm in each direction. For cabbage, interactions between plants appeared to be dominated by the requirement for sufficient space to deploy the shoots for efficient light interception. (+info)