Bilirubin: an endogenous scavenger of nitric oxide and reactive nitrogen species

Abstract

Bilirubin is the final product of heme metabolism. Until recently, bilirubin was considered as a mere by-product of heme degradation but, in the last 20 years, many papers have appeared in the literature demonstrating that this bile pigment is endowed with a strong antioxidant activity, being able to counteract the cellular damage elicited by reactive oxygen species in many in vitro and in vivo experimental systems. Interestingly, compelling evidence has shown that BR can serve as an endogenous scavenger of both nitric oxide and reactive nitrogen species, thus widening the protective role of bilirubin to other reactive species originating within the cellular milieu. The aim of this paper is to give an overview of the interaction between bilirubin and nitric oxide/reactive nitrogen species; furthermore, the possible pathophysiological and clinical relevance of this interaction will be discussed.     

Dipyrone and aminopyrine are effective scavengers of reactive nitrogen species

Abstract

Reactive nitrogen species (RNS), namely nitric oxide (NO^?) and peroxynitrite (ONOO^?) are produced in the inflammatory sites and may contribute to the deleterious effects of inflammation. The aim of the present study was to evaluate the putative scavenging effect of a particular group of non-steroidal anti-inflammatory drugs (NSAIDs), the pyrazolone derivatives dipyrone, aminopyrine, isopropylantipyrine, and antipyrine against RNS, using in vitro non-cellular screening systems. The results obtained showed that dipyrone and aminopyrine were highly potent scavengers of NO^? and ONOO^? while antipyrine exerted little effect and isopropylantipyrine no effect whatsoever against these two RNS and that, in the presence of bicarbonate, the scavenging potencies of both dipyrone and aminopyrine were slightly decreased. It could thus be inferred that the observed scavenging effects may be of therapeutic benefit for patients under anti-inflammatory treatment with dipyrone and aminopyrine in the case of overproduction of RNS. On the other hand, the possible depletion of physiological NO^? concentrations, namely at the gastrointestinal tract as well as the formation of reactive derivatives of aminopyrine and/or dipyrone, resulting from their reaction with RNS, may otherwise be harmful for these patients.     

NO• chemistry: a diversity of targets in the cell

Abstract

NO^? alone is a poorly reactive species; however, it is able to undergo secondary reactions to form highly oxidizing and nitrating species, NO₂^?, N₂O₃, and ONOO^?. These secondary reactive nitrogen species (RNS) are capable of modifying a diversity of biomolecular structures in the cell. The chemical properties of individual RNS will be discussed, along with their ability to react with amino acids, metal cofactors, lipids, cholesterol, and DNA bases and sugars. Many of the identified RNS-induced modifications have been observed both in vitro and in vivo. Several of these chemical modifications have been attributed with a functional role in the cell, such as the modulation of enzyme activity. Other areas in the field will be discussed, including the ability of RNS to react with metabolites, RNA, and substrates in the mitochondrion, and the cellular removal/repair of RNS-modified structures.     

Biological regulation through protein disulfide bond cleavage

Abstract

It is thought that disulfide bonds in secreted proteins are inert because of the oxidizing nature of the extracellular milieu. We have suggested that this is not necessarily the case and that certain secreted proteins contain one or more disulfide bonds that can be cleaved and that this cleavage is central to the protein's function. This review discusses disulfide bond cleavage in the secreted soluble protein, plasmin. Cleavage of plasmin disulfide bond(s) triggers peptide bond cleavage and formation of the tumour angiogenesis inhibitor, angiostatin. Tumour cells secrete phosphoglycerate kinase which facilitates cleavage of the plasmin disulfide bond(s). Phosphoglycerate kinase is not a conventional disulfide bond reductase. We propose that phosphoglycerate kinase facilitates cleavage of a particular plasmin disulfide bond by hydroxide ion, which results in formation of a sulfenic acid and a free thiol. The free thiol is then available to exchange with another nearby disulfide bond resulting in formation of a new disulfide and a new free thiol. The reduced plasmin is then susceptible to discreet proteolysis which results in release of angiostatin.     

Lanthanum chloride bidirectionally influences calcification in bovine vascular smooth muscle cells

Vascular calcification (VC) is frequent prevalence in patients with chronic kidney disease (CKD) and atherosclerosis. Lanthanum carbonate is used as an orally administered phosphate-binding agent to reduce the gastrointestinal absorption of phosphate and ameliorate VC in advanced CKD. In this study, we used bovine vascular smooth muscle cells as a model VC in vitro and studied the effects of lanthanum chloride on calcium deposition. Exposure of cells to LaCl(3) at the concentration of 0.1 μM suppressed the β-glycerophosphate-induced alkaline phosphatase activity and calcium deposition. Furthermore, LaCl(3) upregulated the β-glycerophosphate-suppressed expression of calcium-sensing receptor. In contrast to the inhibitory effect of LaCl(3) on calcium deposition, higher level lanthanum (50 μM) was found to promote immediately precipitation of calcium phosphate in cell culture medium. At this concentration, LaCl(3) was found to induce cell apoptosis which involves caspases-9 and -3. These data indicate that the promotory effect of LaCl(3) on calcium deposition is likely mediated by induction of apoptosis. Our in vitro findings do suggest that, in the context of raised lanthanum, greater attention should be paid to potential toxic effects associated to the use of lanthanide-based drugs.     

The misapplication of Hedwigia integrifolia P. Beauv. and identity of Gymnostomum imberbe Sm. (Hedwigiaceae,Bryopsida)

Abstract

We conducted a 6 year field experiment in an evergreen Quercus ilex forest where we simulated the increased drought projected by Global Circulation Models (GCM) and ecophysiological models for the immediate decades. We tested the hypothesis that enhanced drought will change C, N, P, K, Ca, Fe, Mg, Mo and S concentrations of the widespread moss Hypnum cupressiforme Hedw. and its capacity to absorb nutrients of soil-borne or airborne origin. During the period of study, from 1999 to 2005, the soil moisture in the drought treatment was on average 9% lower than the soil moisture in the control plots. Drought increased the K concentration by 10% and the C concentration by 6%, and decreased the Fe and Mo concentrations by 33% and 18%, respectively, and the N/P content ratio by 15%. A principal component analysis showed that 69% of the variation in moss elemental concentrations is explained by the drought treatment. Drought increased the enrichment factors of several elements, mainly of P, K, Ca, Mg, S and Mo, relative to bedrock extracts, thus showing that the proportion of these elements absorbed from the atmosphere had been increased by drought. The results show that drought increased the concentration of elements linked to drought resistance such as C and K, and decreased the contents of others important for plant productivity such as Mo and Fe. Drought thereby changed moss stoichiometry, and this could also affect the palatability and quite probably, the moss–herbivore relationships and decomposition rates.     

Seasonal phosphatase activities of mosses from Upper Teesdale,northern England

Abstract

Changes in tissue nutrient concentrations and surface phosphatase activities of eight moss species were measured over one year in terrestrial and semi-aquatic environments on Widdybank Fell, Upper Teesdale National Nature Reserve, northern England. Rates of phosphatase activity in apical regions of moss shoots differed markedly between species, but were generally greatest in the winter and least in the summer in most species. Mean values for phosphomonoesterase activity (µmol para-nitrophenol g^-1 d.wt h^-1) ranged from 18.2 for Polytrichum commune to 85.8 for Palustriella commutata var. falcata. Mean phosphodiesterase activity ranged from 3.1 for Polytrichum commune to 86.2 for Hylocomium splendens. In contrast, tissue nitrogen and phosphorus concentrations remained relatively constant throughout the year. Phosphatase activities were negatively correlated with tissue phosphorus concentration for several species, although few relationships were detected between ambient nutrient concentrations and phosphatase activity, tissue nitrogen, or tissue phosphorus concentration. These results demonstrate that phosphatase activities can provide a sensitive indicator of nutrient stress in terrestrial and semi-aquatic mosses, notably in the ectohydric Hylocomium splendens. However, further studies at sites with a wide range of nutrient levels are required to determine whether the technique can be used to indicate ambient nutrient status.     

Contrasting effects of ammonium enrichment on fen bryophytes

Abstract

In the second half of the 20th century, frequency and diversity of brown mosses strongly declined in Dutch fens while species of Sphagnum and Polytrichum increased markedly. We hypothesized that high ammonium (NH₄⁺) concentrations in Dutch precipitation have promoted these species changes. To test this hypothesis, we examined the effects of varying concentrations (1–1000 μM) of NH₄⁺ on a brown moss (Calliergonella cuspidata), two species of Sphagnum (S. contortum and S. squarrosum) and Polytrichum commune in a hydroponic experiment. A second goal was to identify possible interspecific differences in the mechanisms responsible for NH₄⁺ detoxification. Dry matter production and tissue cation concentrations indicated decreasing sensitivity to NH₄⁺ in the order C. cuspidata>S. contortum>S. squarrosum and P. commune. In S. squarrosum and P. commune, the highest NH₄⁺ treatments induced strong increases in tissue nitrogen (N) and amino acid concentrations. Sphagnum contortum showed a similar, but less marked response. In contrast, C. cuspidata showed a negative relationship between external NH₄⁺ concentrations and both tissue N and amino acid concentrations (except arginine). Our findings support the hypothesis that the present rates of NH₄⁺ deposition in The Netherlands are detrimental to brown mosses, but not to fast-growing Sphagnum and Polytrichum species. The efficiency of the NH₄⁺ assimilation apparatus, producing N-rich amino acids, probably plays a key role in determining sensitivity to increased NH₄⁺ deposition.     

Expansion of Sphagnum fallax in bogs: striking the balance between N and P availability

Abstract

Nitrogen deposition may cause shifts in the Sphagnum species composition of bogs, ultimately affecting the conservation value of these systems. We studied the effects of N and P on the expansion of S. fallax and S. flexuosum in bogs. We related historical census data of S. fallax, S. flexuosum, and four of their accompanying species to changes in N deposition. In addition, we conducted two fertilization experiments with N and P; one at a low deposition site with S. flexuosum and one at two high deposition sites with S. fallax. Finally, we related existing data on capitulum N and P concentrations of S. fallax to its abundance in the field.

A relative increase in observed frequency of S. fallax coincided with an historical increase in N deposition in the Netherlands. There was no indication that S. fallax consistently outcompeted one of the other five Sphagnum species; the observed frequency of the Sphagnum species analysed was rather stable with time. The census data on S. flexuosum did not indicate a response to N deposition, but the species expanded at the low N deposition site when extra N was applied. In contrast, the expansion of S. fallax at the high deposition sites was limited by P. Organic nutrient concentrations suggested that when S. fallax can maintain a capitulum N concentration of 7 mg g^-1 or higher and a P concentration of 0.7 mg g^-1 or higher the species can grow to dominate.

We conclude that S. fallax will gradually colonize an increasing number of new habitats in areas with a low, albeit increasing, N deposition, but will only grow to dominate when P supply is adequate. Then, the expansion of S. fallax may lead to ousting of the other Sphagnum species present.