Plant hormone homeostasis and the control of avocado fruit size

Control of plant hormone homeostasis is crucial for normal organdevelopment in plants. To elucidate the contribution of plant hormonehomeostasis to fruit growth, tissue distribution and activity of xanthinedehydrogenase (XDH), abscisic aldehyde (AB-ald)- and indole acetaldehyde(IA-ald) oxidase, and cytokinin oxidase (CKOX) were determined in seed, seedcoat and mesocarp of normal 'Hass avocado and its small-fruitphenotype during the linear phase of growth. Activity of these enzymes wasrelated to the tissue content of indole-3-acetic acid (IAA) and abscisic acid(ABA). IA-ald oxidase was present only in seed tissue whereas AB-ald oxidase andXDH activity was found in seed and mesocarp tissue. Seed of the small'Hass fruit had increased XDH and AB-ald oxidase activity and highendogenous ABA, but reduced IA-ald oxidase activity and adenine. There was nodifference in seed, seed coat and mesocarp CKOX activity between normal andsmall fruit. Inhibition of XDH activity in whole fruit by treatment withallopurinol decreased IAA and increased ABA of seed tissue. In mesocarp ofripening fruit allopurinol increased ABA and IAA but had no effect on levels ofiP. Results indicate that activity of IA-ald and AB-ald oxidases in avocadofruit contribute to maintenance of the IAA/ABA ratio in seed and mesocarp tissueand that increased AB-ald oxidase, or reduced IA-ald oxidase, may be part of thesyndrome associated with the appearance of a small-fruit phenotype.     

Xanthine accumulation and vacuolization inChlamydomonas reinhardtii cells

Summary Utilization of xanthine as the sole nitrogen source for growth byChlamydomonas reinhardtii cells involved the formation of a transient, intracellular pool of xanthine. Up to 20% of the total xanthine supplied to the medium was not assimilated after uptake but stored in the cells at concentrations that exceeded xanthine solubility in water. At the subcellular level, a massive accumulation of starch grains in the chloroplast and the appearance of many vacuoles in the cytoplasm distinguished xanthine-grown from ammonium-grown cells. Starch accumulation, but not development of vacuoles, was also observed in N-starved cells. Uptake experiments with radio-labelled xanthine showed that this accumulates only in the cytoplasm, most probably inside vacuoles. The electron-dense material observed in vacuoles of xanthine-grown cells suggests that the intracellular xanthine is in part solid xanthine.     

Improved method for measurement of human plasma xanthine oxidoreductase activity

The XOR activity in human plasma was measured by quantifying the XOR-derived uric acid (UA) in plasma using the high-performance liquid chromatography (HPLC) equipped with a UV detector. Chromatographic separation consisted of the mobile phase (a mixture of 0.1% trifluoroacetic acid in Milli-Q water and 0.085% trifluoroacetic acid in acetonitrile in a mix ratio of 99:1) running through a Zorbax StableBond SB-C(18) column at a flow-rate of 1 ml/min. Deproteinization with heat-treatment of plasma samples after the reaction was used in the assay to avoid splitting of the UA and xanthine peaks caused by acid deproteinization that could interfere the accurate determination of human plasma XOR activity in our case. Based on the examination of the dependence of XOR activity on added amounts of xanthine and reaction times, the amount of xanthine and reaction time for XOR activity assay were determined to prevent the errors caused by the limiting effect of substrates and plateau phase of the reaction. Using this method, human plasma XOR activities of 25 healthy people were measured. The average human plasma XOR activity was 2.1+/-0.8 (x10(-3) U/ml).     

The effect of rebamipide on gastric xanthine oxidase activity and type conversion in ethanol-treated rats

Rebamipide, a novel antipeptic ulcer drug, 2-(4-chlorobenzoylamino)-3-[2(1H)-quinolinone-4-yl]-propionic acid, was studied for its inhibitory effect on gastric xanthine oxidase activity and type conversion of the enzyme that has a profound role in free radical generation. Intraperitoneal administration of rebamipide at 60 mg/kg body weight reduced gastric mucosal hemorrhagic lesions and lipid peroxidation, which was proportional to the inhibitory effect of rebamipide on alcohol-induced xanthine oxidase-type conversion and enzyme activity. It was also observed that the activity of xanthine oxidase was significantly inhibited by administration of rebamipide at 60 mg/kg body weight, leading to a significant reduction of lipid peroxide content in alcohol-treated rats. The results suggest that alcohol-induced gastric mucosal lesions might be, in part, due to the increased activity of xanthine oxidase and type conversion rate of the enzyme and the protective effect of rebamipide on gastric mucosal lesions would result from its ability to protect against oxidative stress on gastric mucosal lesions of alcohol-treated rats.     

Characterization of aldehyde oxidase from <Emphasis Type="Italic">Brevibacillus</Emphasis> sp. MEY43 and its application to oxidative removal of glutaraldehyde

Bacterium MEY43, an isolate from soil, produced aldehyde oxidase when it was cultivated in a medium containing methanol as a sole source of carbon and energy. The methylotrophic bacterium was identified as Brevibacillus sp. Its cultivation in media containing other substrates, such as ethanol and glucose, resulted in little production of the enzyme. Aldehyde oxidase purified from a cell-free extract of the bacterium was a hetero-trimeric protein comprised of large, medium, and small subunits with molecular masses of 87, 35, and 19 kDa, respectively. Its UV/visible spectrum and the presence of molybdenum, 5′-CMP, flavin adenine dinucleotide, iron, and acid-labile sulfur suggested that the enzyme belonged to the xanthine oxidase family. The enzyme acted on a wide range of aliphatic and aromatic aldehydes. The K _m value for formaldehyde was 32 mM, whereas those for the other aldehydes tested were below 0.2 mM. When 10 mM glutaraldehyde was treated with 2.0 units of the enzyme ml⁻¹ in the presence of 100 units ml⁻¹ catalase for 120 min, the concentration of the aldehyde decreased to below a detectable level.     

Xanthine oxidoreductase: A journey from purine metabolism to cardiovascular excitation-contraction coupling

Xanthine oxidoreductase (XOR) is a ubiquitous complex cytosolic molybdoflavoprotein which controls the rate limiting step of purine catabolism by converting xanthine to uric acid. It is known that optimum concentrations of uric acid (UA) and reactive oxygen species (ROS) are necessary for normal functioning of the body. The ability of XOR to perform detoxification reactions, and to synthesize UA and reactive oxygen species (ROS) makes it a versatile intra- and extra-cellular protective “housekeeping enzyme”. It is also an important component of the innate immune system. The enzyme is a target of drugs against gout and hyperuricemia and the protein is of major interest as it is associated with ischemia reperfusion (I/R) injury, vascular disorders in diabetes, cardiovascular disorders, adipogenesis, metabolic syndrome, cancer, and many other disease conditions. Xanthine oxidoreductase in conjugation with antibodies has been shown to have an anti-tumor effect due to its ability to produce ROS, which in turn reduces the growth of cancer tissues. Apart from this, XOR in association with nitric oxide synthase also participates in myocardial excitation-contraction coupling. Although XOR was discovered over 100 years ago, its physiological and pathophysiological roles are still not clearly elucidated. In this review, various physiological and pathophysiological functional aspects of XOR and its association with various forms of cancer are discussed in detail.     

RNA interference-mediated suppression of xanthine dehydrogenase reveals the role of purine metabolism in drought tolerance in Arabidopsis

We have previously demonstrated that RNA interference-mediated suppression of xanthine dehydrogenase (XDH), the rate-limiting enzyme in purine degradation, causes defects in the normal growth and development of Arabidopsis thaliana. Here, we investigated a possible role for XDH in drought tolerance, since this enzyme is also implicated in plant stress responses and acclimatization. When XDH-suppressed lines were subjected to drought stress, plant growth was markedly reduced in conjunction with significantly enhanced cell death and H₂O₂ accumulation. This drought-hypersensitive phenotype was reversed by pretreatment with exogenous uric acid, the catalytic product of XDH. These results suggest that fully functional purine metabolism plays a role in the Arabidopsis drought acclimatization.     

Endophytic fungi from <Emphasis Type="Italic">Nerium oleander</Emphasis> L (Apocynaceae): main constituents and antioxidant activity

Diverse endophytic fungi exist within plant aerial tissues, with a global estimate of up to a million undescribed species. These endophytes constitute a rich bio-resource for exploration to discover new natural products. Here we investigate fungal endophytes associated with a medicinal plant, Nerium oleander L. (Apocynaceae). A total of 42 endophytic fungal strains were isolated from the host plant. Total antioxidant capacity, xanthine oxidase inhibitory activity, antimicrobial activity, and total phenolic content (TPC) were evaluated for 16 representative fungal cultures grown in improved Czapek’s broth and for the host plant. The total antioxidant capacities and phenolic contents of the fungal cultures ranged from 9.59 to 150.79 μmol trolox/100 mL culture, and from 0.52 to 13.95 mg gallic acid/100 mL culture, respectively. The fungal culture of an endophytic strain Chaetomium sp. showed the strongest antioxidant capacity, contained the highest level of phenolics, and to some extent inhibited xanthine oxidase activity with an IC₅₀ value of 109.8 μg/mL. A significant positive correlation was found between antioxidant capacity and TPC in the tested samples. Most of the endophytic fungal cultures tested have a wide range of antimicrobial activities, which were not very strong, but much better than those of the host plant. The major bioactive constituents of the fungal cultures were investigated using LC-ESI-MS and GC-MS, and preliminary identification detected phenolics (e.g. phenolic acids and their derivatives, flavonoids) and volatile and aliphatic compounds. This study shows that the endophytic fungi isolated from N. oleander can be a potential antioxidant resource.     

Synthesis of resveratrol analogues, and evaluation of their cytotoxic and xanthine oxidase inhibitory activities

Thirteen resveratrol (=5-[(E)-2-(4-hydroxyphenyl)ethenyl]benzene-1,3-diol) analogues with a CHO group have been prepared by partial synthesis from resveratrol. The synthesized compounds have been evaluated for their cytotoxic activity against a human nasopharyngeal epidermoid tumor cell line KB, as well as for their xanthine oxidase inhibitory activity. Compounds 2, 3, and 6a showed the most significant cytotoxic activities against the cell line KB, and compound 2 also exhibited strong xanthine oxidase inhibitory activity.     

Sodium nitrite downregulates vascular NADPH oxidase and exerts antihypertensive effects in hypertension

Dietary nitrite and nitrate are important sources of nitric oxide (NO). However, the use of nitrite as an antihypertensive drug may be limited by increased oxidative stress associated with hypertension. We evaluated the antihypertensive effects of sodium nitrite given in drinking water for 4 weeks in two-kidney one-clip (2K1C) hypertensive rats and the effects induced by nitrite on NO bioavailability and oxidative stress. We found that, even under the increased oxidative stress conditions present in 2K1C hypertension, nitrite reduced systolic blood pressure in a dose-dependent manner. Whereas treatment with nitrite did not significantly change plasma nitrite concentrations in 2K1C rats, it increased plasma nitrate levels significantly. Surprisingly, nitrite treatment exerted antioxidant effects in both hypertensive and sham-normotensive control rats. A series of in vitro experiments was carried out to show that the antioxidant effects induced by nitrite do not involve direct antioxidant effects or xanthine oxidase activity inhibition. Conversely, nitrite decreased vascular NADPH oxidase activity. Taken together, our results show for the first time that nitrite has antihypertensive effects in 2K1C hypertensive rats, which may be due to its antioxidant properties resulting from vascular NADPH oxidase activity inhibition.