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Immunogold labelling revealed the presence of lipoxygenase (LOX) in different parts and types of anther cells of Gagea lutea. LOX was found in the cytoplasm and close to ER elements in epidermal and endothecial cells, and close to the cell walls of the latter. The positive immunoreaction to LOX was less intense in the middle layers and the loculus of the anther, where single immunogold particles were concentrated at the cell walls of these layers and in the protoplast masses, in vacuoles, close to mitochondria, inside plastids, and in the liquid of the anther cavity. LOX occurred in the cytoplasm and around ER elements of pollen grains as well as in the exine layer, particularly in contact regions between the outer and inner exine layers. The correlations between LOX localization in different anther cells and the functioning of particular anther parts are discussed.
Lipid peroxidation, which is a natural and essential process, can occur in a non-enzymatic and/or enzymatic way in plant cells. Some of its products have cytotoxic effects on cells, but others function as plant effectors. The lipid peroxidation in plants exposed to heavy metal stress depends on the metal, plant organ, plant species and its genotype.
The activity of lipoxygenase (EC 1.13.11.12) in Arabidopsis thaliana (L.) Heynh seedlings and mature plants was estimated spectrophotometrically at 234 nm. Linoleic acid was used as a substrate. Lipoxygenase activity showed two pH optima: at 7.0 and 10.0 in seedlings, and at pH 8.0 and 10.0 in leaves of mature plants. Seven-week-old plants were transferred to a hydroponic system and treated with different concentrations of Cd2+ or Cu2+ [in μM]: 0, 5, 25, 50, 100 for 7 days. The lipoxygenase activities at pH 8.0 and 10.0 depended on the metal that was added to the nutrient solution. The main change in lipoxygenase activity was under Cd2+ stress at pH 8.0 and under Cu2+ excess at pH 10.0.
Nitric oxide (NO) is a potent extracellular and intracellular physiological messenger. However, NO liberated in excessive amounts can be involved in macromolecular and mitochondrial damage in brain aging and in neurodegenerative disorders. The molecular mechanism of its neurotoxic action is not fully understood. Our previous data indicated involvement of NO in the release of arachidonic acid (AA), a substrate for cyclo- and lipoxygenases (COX and LOX, respectively). In this study we investigated biochemical processes leading to cell death evoked by an NO donor, sodium nitroprusside (SNP). We found that SNP decreased viability of pheochromocytoma (PC12) cells in a concentration- and time-dependent manner. SNP at 0.1 mM caused a significant increase of apoptosis-inducing factor (AIF) protein level in mitochondria. Under these conditions 80% of PC12 cells survived. The enhancement of mitochondrial AIF level might protect most of PC12 cells against death. However, NO released from 0.5 mM SNP induced massive cell death but had no effect on protein level and localization of AIF and cytochrome c. Caspase-3 activity and poly(ADP-ribose) polymerase-1 (PARP-1) protein levels were not changed. However, PARP activity significantly decreased in a time-dependent manner. Inhibition of both COX isoforms and of 12/15-LOX significantly lowered the SNP-evoked cell death. We conclude that AIF, cytochrome cand caspase-3 are not responsible for the NO-mediated cell death evoked by SNP. The data demonstrate that NO liberated in excess decreases PARP-1 activity. Our results indicate that COX(s) and LOX(s) are involved in PC12 cell death evoked by NO released from its donor, SNP.
The effect of exogenous salicylic acid (SA) on the vase life of cut rose flowers was investigated. 'Yellow Island' roses were obtained from a commercial grower. The roses were pulse treated with SA (0, 50, 100, 150, 200 mg l-1) for 18 hours. Com­pared to the control treatment (DW), the greatest delay in senescence was obtained in cut roses treated with 150 mg l-1 SA. With this treatment, flower vase life was dou­bled. The physiological characteristics such as protein concentration, lipid peroxida- tion (MDA) level and enzyme activity such as lipoxygenase (LOX), superoxide dis- mutase (SOD) and peroxidase (POD) were measured only in the control (DW) and those kept in a solution of 150 mg l-1 salicylic acid (SA) for over 8 days of vase life. Degradation of protein and accumulation of MDA during vase life was shown to be suppressed by 150 mg l-1 SA. The activity of LOX and POD gradually increased with progressing flower senescence, while SOD activity declined. The flowers which had been pulsed with SA exhibited lower LOX and POD activity, and a higher activity of SOD. Overall, the results suggest that SA increases vase life by improving the anti- oxidant system and reducing oxidative stress damages during rose flower senescence.
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Leukotrienes in mucosal damage and protection

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Exposure of the rat gastric mucosa to ethanol stimulates the generation of leukotriene (LTC₄) and 15-hydroxyeicosatetraenoic acid, hut not of thromboxanes and prostaglandins. Lipoxygenase activation is not found with other topical irritants or nonsteroidal anti-inflammatory drugs. A number of gastroprotective drugs dose-dependently inhibit the stimulatory action of ethanol on mucosal LTC₄ formation closely parallel to their protective activity suggesting that ethanol-induced damage and activation of lipoxygenases may involve common targets which are simultaneously counteracted by certain types of protective agents. Selective inhibition of 5-lip- oxygenase, however, does not confer protection against gastric mucosal damage caused by topical irritants or non-steroidal anti-inflammatory drugs. Thus, although leukotrienes may mediate certain reactions elicited by gastric ulcerogens such as submucosal venular constriction and mucosal micro vascular engorgement, they do not appear to be major mediators of ulcerogen-induced tissue necrosis. The contribution of other products of the various pathways of arachidonic acid metabolism to gastric mucosal injury and the mechanism underlying the close interrelationship between protection and inhibiton of LTC₄ formation observed with certain compounds remains to be investigated.
Lipoxygenase was localized in the primary leaves of Phaseolus coccineus (L.), seedlings treated with 25 µM Cd and in control plants using the immunogold method. The enzyme was localized mainly in the peripheral parts of protoplast of control plant cells. It was found in the cell wall, along the ER elements, at plastid lamellae and inside the mitochondria. In Cd-treated seedlings the elements of parenchyma cells showed an atypical inner structure. The immunolabelling of LOX was less intensive in comparison with control. The enzyme was found in the cytoplasm, at the cell wall area, vacuoles and in the plastid stroma as single gold particles. LOX activity optima were determined at pH 7.0 and 8.0 for both linoleic and linolenic acid used as substrates. After 2 days of seedlings exposure to Cd the activity of LOX decreased at pH 7.0 and 8.0 when linoleic acid was used as substrate, and strongly declined at pH 7.0 after 4 days of the metal treatment. When linolenic acid was the substrate LOX activity slightly increased after 2 days of the plants exposure to Cd, but after 4 days it rapidly decreased at pH 7.0. The changes in LOX activity are discussed.
The experiment studied the synergetic effect of casein and its enzymatic hydrolysate in combination with ascorbic acid and beta-carotene in slowing down the oxidation of linoleic acid in model oxidation systems. The obtained results indicate that the antioxidant efficiency of applied antioxidants depends on the type of the system in which the lipid oxidation occurs and from the mechanism of oxidation (autooxidation, photosensitization, oxidation by lipooxygenase). For the applied preparations the best antioxidant activities were obtained for emulsion system of linoleic acid in the reaction of the autooxidation, where the co-operation between antioxidants of different physical properties and mechanisms of the antioxidant action in various emulsion phases was possible
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