Toxicity of three rare earth elements, and their combinations to algae, microcrustaceans, and fungi.

Rare earth elements (REEs) are naturally distributed in the environment, and are increasingly being used in agriculture and high technology materials worldwide, thereby increasing anthropogenic contamination and environmental risks. There exists scarce and contradictory toxicity information about REEs; hence, more studies are required, especially on their mixtures. Thus, this study aimed to assess the toxicities of La3+, Nd3+, Sm3+, and the combinations of these elements (binary 1:1 and ternary 1:1:1), to organisms from different trophic levels: producers (the microalgae Chlorella vulgaris and Raphidocelis subcapitata), primary consumers (the microcrustaceans Daphnia similis and Artemia salina), and decomposers (the fungi Penicillium simplicissimum and Aspergillus japonicus). Ecotoxicological bioassays were performed, and toxic concentrations were determined. Thereafter, toxicities of single and mixture REEs were classified as slightly to highly toxic according to their toxic units. Finally, a concentration addition (CA) model was used to estimate how REEs interact upon combining. Nd3+ was the most toxic element for all organisms, especially D. similis (48 h LC50 9.41 mg.L-1), and was therefore classified as highly toxic. Sm3+ promoted cell agglomeration in Chlorella vulgaris and was the most toxic of the tested elements for this organism (72 h IC50 25.78 mg.L-1). The CA model revealed synergistic responses for most of the combinations, principally Nd3+ + Sm3+, which was the most toxic combination for the tested organisms. Both fungi were the most resistant organisms, and A. japonicus produced exudate and sclerotia, which help in the detoxification of chemicals. Owing not only to the fact that fungi displayed a higher resistance to REEs, but also due to the absence of regulations for REEs released from the agricultural or industrial sector, and the lack of methods to treat effluents or to dispose of technological items containing REEs, these organisms should be considered as a model for the biosorption or bioremediation of REEs. Finally, the toxic effects of REEs, particularly Nd3+, on the biota and human health should be the focus of future studies due to their increased use in technology.

Endothelium-dependent relaxations to adenosine in juvenile rabbit pulmonary arteries and veins.

We studied the actions of adenosine and its analogues 5'-(N-ethylcarboxamido)-adenosine (NECA) and N6-cyclohexyladenosine (CHA) in pulmonary vessels isolated from juvenile rabbits. Pulmonary arteries relaxed in a concentration-dependent fashion to all three compounds. Pretreatment with the methylxanthine 8-p-sulfophenyltheophylline shifted the concentration-response curves to adenosine and NECA rightward, indicating that the vasodilator effects were mediated by the adenosine receptor. The order of potency of adenosine compounds was NECA > adenosine > CHA, indicating that the A2-receptor mediates relaxations to adenosine in rabbit pulmonary arteries. Endothelium rubbing attenuated relaxations to adenosine at concentrations of < or = 3 x 10(-7) M and to all NECA concentrations. Inhibition of nitric oxide synthase with NG-nitro-L-arginine (L-NNA) similarly attenuated relaxations at concentrations of < or = 3 x 10(-7) M for adenosine and < or = 3 x 10(-8) M for NECA. With the use of the same methods, a substantial endothelial contribution was additionally observed in pulmonary veins to the vasodilator effects of NECA. We conclude that adenosine, and the more specific A2-receptor agonist NECA, dilate pulmonary arteries and veins isolated from young rabbits via a mechanism that is partially dependent on endothelium-derived nitric oxide.

Developmental differences in endothelium-dependent responses in isolated ovine pulmonary arteries and veins.

Despite evidence for an important role for endothelium-derived relaxing factor (EDRF) in transitional circulation, previous in vitro studies of newborn pulmonary arteries have demonstrated diminished EDRF activity when compared with arteries from older animals. We studied pulmonary arteries and veins isolated from early newborn and juvenile sheep using standard tissue bath techniques. Incubation of vessels with the nitric oxide synthase inhibitor N omega-nitro-L-arginine (L-NNA) constricted veins but not arteries from both age groups. Further studies using preconstricted vessels revealed that arteries relaxed to acetylcholine (ACh), with significantly greater responses observed in juvenile arteries. Veins from both age groups contracted to ACh. Pretreatment with prostaglandin inhibitors (indomethacin and SQ 29,548) diminished ACh relaxations in pulmonary arteries from both age groups, greatly enhanced relaxations to ACh in newborn pulmonary veins, and depressed contractions in juvenile pulmonary veins. Removal of endothelium mechanically or functionally with prostaglandin inhibitors and L-NNA eliminated relaxations to ACh in pulmonary arteries from both age groups and resulted in contractions in veins. We conclude that isolated pulmonary veins from newborn sheep exhibit both baseline and stimulated release of EDRF, and we speculate that these venous responses may be important in the transitional pulmonary circulation.

Characterization and mechanisms of H2O2-induced contractions of pulmonary arteries.

We studied H2O2-induced contractions of isolated rabbit intrapulmonary arteries mounted in standard tissue baths. All vessels were pretreated with a thromboxane A2/prostaglandin H2 receptor antagonist, SQ 29,548, to block immediate transient contractions to H2O2 and to isolate slowly developing sustained contractions. When exposed to H2O2 (0.1, 0.2, 0.3, 0.6, and 1.0 mM) for 30 min, vessels contracted in (0.1, 0.2, 0.3, 0.6, and 1.0 mM) for 30 min, vessels contracted in a concentration-dependent fashion between 0.1 and 0.3 mM H2O2; contractions at 0.6 and 1.0 mM H2O2 were not significantly different from those at 0.3 mM H2O2. During recovery (90 min) from H2O2 exposures, baseline tension was significantly greater, but active tension (10 microM phenylephrine) was significantly less for vessels previously exposed to 0.6 and 1.0 mM H2O2. Contractions to 0.3 mM H2O2 were not blunted by the following interventions: 1) endothelium rubbing, 2) incubation in Ca(2+)-free 100 microM ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA) Krebs-Ringer solution, 3) incubation in the Ca(2+)-free solution and depletion of ryanodine (20 microM)-sensitive Ca(2+) stores, or 4) pretreatment with the protein kinase C inhibitor 1-(5-isoquinolinylsulfonyl)-3-methyl-piperazine (20 microM). However, contractions were depressed by approximately 50% when vessels were pretreated with the phospholipase C/serine esterase inhibitor 2-nitro-4-carboxy-phenyl-N,N-diphenylcarbamate (50 microM). These results suggest that slow-developing contractions to H2O2 are concentration dependent and may result, in part, from activation of a serine esterase(s) and/or phospholipase C.

Neuronal type N calcium channels mediate norepinephrine release in isolated rabbit pulmonary artery.

Several types of voltage sensitive calcium channels (VSCC) including types N and Ln are present in varying degrees in neurons from different locations and they appear to differ from the Lm type VSCC on muscle cells. Omega conotoxin blocks N and Ln channels but not Lm channels. Verapamil blocks Ln and Lm but not N channels. The purpose of this study was to determine whether N or Ln channels mediate norepinephrine (NE) release in isolated rings of rabbit pulmonary artery. The release of endogenous NE from intramural sympathetic neurons was evoked by field electrical stimulation (ES) of the tissues. Conotoxin (10(-6) M) had no effect on contractions of rabbit pulmonary artery induced by 40 mM K+ or exogenous NE (10(-9)-10(-5) M) but significantly depressed the response to ES (21V, 0.5 msec pulse duration) at all stimulation frequencies (1-20Hz). Thus, the effects of conotoxin are restricted to the N and/or Ln calcium channels on neurons in this issue. Verapamil (5 x 10(-6) M) depressed 40 mM K(+)-induced contractions by 86 +/- 2% as well as contractions induced by endogenous (ES at 1-20 Hz) and exogenous NE (10(-9)-10(-5) M). Moreover, for contractions of equal magnitude, verapamil depressed the response to endogenous (ES) and exogenous NE to the same degree. Had Ln channels mediated NE release in the tissue, verapamil should have had a greater effect on ES-induced contractions because ES activates both pre- and post-junctional calcium channels whereas exogenous NE activates only post-junctional channels on smooth muscle cells. Thus, it appears that N-type calcium channels mediate NE release in rabbit pulmonary artery.