Editorial

The voltage dependence of the rat renal type II Na⁺/P_i cotransporter (NaP_i-2) was investigated by expressing NaP_i-2 in Xenopus laevis oocytes and applying the two-electrode voltage clamp. In the steady state, superfusion with inorganic phosphate (P_i) induced inward currents (I_p) in the presence of 96 mM Na⁺ over the potential range −140 ≤ V ≤ +40 mV. With P_i as the variable substrate, the apparent affinity constant (K _m ^Pi) was strongly dependent on Na⁺, increasing sixfold for a twofold reduction in external Na⁺. K _m ^Pi increased with depolarizing voltage and was more sensitive to voltage at reduced Na⁺. The Hill coefficient was close to unity and the predicted maximum I_p (I_pmax) was 40% smaller at 50 mM Na⁺. With Na⁺ as the variable substrate, K _m ^Na was weakly dependent on both P_i and voltage, the Hill coefficient was close to 3 and I_pmax was independent of P_i at −50 mV. The competitive inhibitor phosphonoformic acid suppressed the steady state holding current in a Na⁺-dependent manner, indicating the existence of uncoupled Na⁺ slippage. Voltage steps induced pre–steady state relaxations typical for Na⁺-coupled cotransporters. NaP_i-2-dependent relaxations were quantitated by a single, voltage-dependent exponential. At 96 mM Na⁺, a Boltzmann function was fit to the steady state charge distribution (Q-V) to give a midpoint voltage (V_0.5) in the range −20 to −50 mV and an apparent valency of ∼0.5 e⁻. V_0.5 became more negative as Na⁺ was reduced. P_i suppressed relaxations in a dose-dependent manner, but had little effect on their voltage dependence. Reducing external pH shifted V_0.5 to depolarizing potentials and suppressed relaxations in the absence of Na⁺, suggesting that protons interact with the unloaded carrier. These findings were incorporated into an ordered kinetic model whereby Na⁺ is the first and last substrate to bind, and the observed voltage dependence arises from the unloaded carrier and first Na⁺ binding step.     

Variations in the natural abundance of 15N in ryegrass/white clover shoot material as influenced by cattle grazing

Biological N2 fixation in clover is an important source of N in low external-N input farming systems. Using the natural 15N-abundance method, variations in N2 fixation were investigated in grazed and mowed plots of a ryegrass/white clover field. Ryegrass 15N varied considerably, from 0.2 to 5.6 under mowed conditions and from –3.3 to 11.6 under grazed conditions. Variations in 15N white clover were lower than in ryegrass, especially in the mowed plots (SE = 0.05, n = 20). The variations in the percentage of nitrogen derived from the atmosphere (%Ndfa) in white clover were highest in the grazed plots where it ranged from 12 to 96% (mean = 64%) compared with the mowed plots where it ranged from 64 to 92% (mean = 79%). Thus, the N2 fixation per unit white clover DM in the grazed ley was lower and more variable than under mowing conditions.Urine from dairy cows equivalent to 0, 200, 400 and 800 kg N ha-1 was applied to a ryegrass/white clover plot 6, 4 or 2 weeks before harvest. Without urine application 15N of ryegrass was positive. By increasing urine application (15N = –1) two weeks before sampling, the 15N of ryegrass decreased strongly to about –7 (P < 0.001). However, this effect was only observed when urine was applied two weeks before sampling. When applying 800 kg N four and six weeks before sampling, 15N in ryegrass was not significantly different from the treatment without urine application. White clover 15N was unaffected by whatever changes occurred in 15N of the plant-available soil N pool (reflected in 15N of ryegrass). This indicates that within the time span of this experiment, N2 fixation per unit DM was not affected by urine. Therefore, newly deposited urine may not be the main contributing factor to the variation in %Ndfa found in the grazed fields. This experiment suggested that the natural abundance method can be applied for estimating %Ndfa without disturbance in natural animal-grazed systems.     

Metabolism of Polyamines in Transgenic Cells of Carrot Expressing a Mouse Ornithine Decarboxylase cDNA

The metabolisms of arginine (Arg), ornithine (Orn), and putrescine were compared in a nontransgenic and a transgenic cell line of carrot (Daucus carota L.) expressing a mouse Orn decarboxylase cDNA. [¹⁴C]Arg, [¹⁴C]Orn, and [¹⁴C]putrescine were fed to cells and their rates of decarboxylation, uptake, metabolism into polyamines, and incorporation into acid-insoluble material were determined. Transgenic cells showed higher decarboxylation rates for labeled Orn than the nontransgenic cells. This was correlated positively with higher amounts of labeled putrescine production from labeled Orn. With labeled Arg, both the transgenic and the nontransgenic cells exhibited similar rates of decarboxylation and conversion into labeled putrescine. When [¹⁴C]putrescine was fed, higher rates of degradation were observed in transgenic cells as compared with the nontransgenic cells. It is concluded that (a) increased production of putrescine via the Orn decarboxylase pathway has no compensatory effects on the Arg decarboxylase pathway, and (b) higher rates of putrescine production in the transgenic cells are accompanied by higher rates of putrescine conversion into spermidine and spermine as well as the catabolism of putrescine.     

CuZn-Superoxide Dismutase, Extracellular Superoxide Dismutase, and Glutathione Peroxidase in Blood from Individuals Homozygous for Asp90Ala CuZn-Superoxide Dismutase Mutation

Abstract: The Asp⁹⁰Ala CuZn-superoxide dismutase mutation is associated with amyotrophic lateral sclerosis (ALS) in both homo- and heterozygous form. We analyzed antioxidant enzymes in blood from 44 individuals homozygous and 114 individuals heterozygous for the Asp⁹⁰Ala mutation as well as 66 blood relatives carrying the wild-type allele only. Erythrocyte CuZn-superoxide dismutase activity was reduced by 9% in the homozygous individuals, confirming previous findings on a smaller cohort. The specific activity of Asp⁹⁰Ala mutant CuZn-superoxide dismutase in erythrocytes was equal to that of isolated mutant enzyme and slightly higher than that of isolated wild-type enzyme. There was no evidence for the presence of inactive mutant molecules in erythrocytes, and the lower activity is due to the occurrence of fewer active molecules. There were no significant differences between the groups in plasma extracellular superoxide dismutase content, and the erythrocyte glutathione peroxidase activities were virtually identical. Also, there were no differences in these parameters between homozygous individuals with or without ALS. There was no evidence for any association with ALS of a polymorphic extracellular superoxide dismutase mutation, Arg²¹³Gly. The absence of response of the blood antioxidant enzymes to the Asp⁹⁰Ala CuZn-superoxide dismutase mutation does not support the theory that the ALS-linked CuZn-superoxide dismutase mutations cause disease by increased oxidant stress.     

A field study of nitrogen dynamics and spring barley growth as affected by the quality of incorporated residues from white clover and ryegrass

Biofumigation refers to the suppression of soil-borne pests and pathogens by biocidal compounds released by Brassicaceous green manure and rotation crops when glucosinolates (GSLs) in their tissues are hydrolysed. We investigated the effect of environment and ontogeny on the GSL production, and thus biofumigation potential, of eight entries from five Brassica species. The environments included autumn and spring sown field plots (FA and FS) and potted plants grown under ambient conditions (PAM) or in a temperature controlled glasshouse at 20 °C/12 °C (PTC). GSL concentration was measured in the root and shoot tissue at buds-raised, flowering and maturity. Of particular interest was the suitability of the pot-grown plants for screening large numbers of brassicas for GSL production. The type of GSLs present in the tissues and their relative proportions remained relatively constant across environments and at different growth stages, with the exception of an increase in indolyl GSLs in the FS environment suspected of being induced by insect attack. Total GSL concentration generally declined from buds-raised to flowering in all environments, and was lowest at maturity. The exceptions were B. campestris, which had higher GSL concentration at flowering than at buds-raised, and the PTC environment in which most species also showed an increase at flowering. Despite GSL types and their proportions remaining relatively constant, the total GSL concentration in the root and shoot tissue of all entries varied significantly with environment (3–10-fold) and was generally ranked FS>PAM>FA>PTC. Interactions between species and environments meant that the ranking of the Brassica entries for total shoot and root GSL concentration changed with environment. However within three entries from B. napus, the ranking was consistent across the environments. The added effect of environment on phenological development and biomass production further influenced GSL production (the product of GSL concentration and biomass) on a ground area basis. The results suggest that glasshouse environments can be used to determine the types and proportions of GSLs present, and to rank entries within, but not between species for the total concentration in the tissues. However the influence of the environment on both GSL concentration and biomass production suggests that an accurate estimate of GSL production on a ground area basis to assess biofumigation potential will require measurement in the target environment.     

Diet and prey consumption of Antarctic petrels Thalassoica antarctica at Svarthamaren, Dronning Maud Land, and at sea outside the colony

The diet of the Antarctic petrel Thalassoica antarctica was studied during two seasons at Svarthamaren, an inland colony in Dronning Maud Land, Antarctica, and in the pack ice off the coast of Svarthamaren. The most important food (wet mass) at Svarthamaren was crustaceans (67%), fish (29%) and squid (5%); however, individuals collected in the pack ice took mostly fish (87%). The prey composition and lengths of prey are comparable to what has been documented in other studies on this species. Estimates of food consumption by birds breeding at Svarthamaren (ca. 250,000 pairs) suggest that approximately 6500 tonnes of crustaceans, 2800 tonnes of fish and 435 tonnes of squid are consumed during the breeding season. The annual consumptions of these birds are estimated to be 34,100 tonnes of crustaceans, 14,700 tonnes of fish, and 2300 tonnes of squid. Satellite telemetry data indicate that Antarctic petrels from Svarthamaren may fly more than 3000 km during one foraging trip, and thus may cover a huge ocean area to obtain their prey. Received: 1 September 1997 / Accepted: 3 February 1998     

Recommendations for the routine sampling of diatoms for water quality assessments in Europe

Many methods for using diatoms for routine monitoring of water quality have been developed in Europe and, in some countries, these are being used to enforce environmental legislation. In order to facilitate their wider use, particularly with respect to European Union legislation, steps are being taken to harmonize methodology. In this paper, the principles and practice of sampling are described in relation to the main habitat types encountered in Europe. Although details of methods and sampling programmes have to be tailored to particular circumstances and the overall objectives of the monitoring, a number of generalizations can be made. Where available, rocks and other hard surfaces are the preferred substrates and methods for sampling these are described. If such substrata are not available, then introduced ('artificial') substrata have many applications. Various types of introduced substrata can be used successfully, so long as some basic precautions are described. Other types of substrata such as macrophytes and macroalgae may also be useful under certain circumstances, although there is less consensus in the literature on the most appropriate methods, and of the validity of comparisons between indices computed from epiphytic and epilithic communities. When designing surveys, it is recommended that as far as possible, extremes of non-water quality factors (e.g. shade, current speed, etc) are avoided, unless these are characteristic of the system under investigation. Detailed guidelines for sampling epilithon are described. Along with the recommendations for sampling other substrata, it is hoped that these provide a framework that can be adapted to most river types in Europe. This revised version was published online in June 2006 with corrections to the Cover Date.     

c-kit immunoreactive interstitial cells of Cajal in the human small and large intestine

 c-kit immunohistochemistry was performed on unfixed frozen sections of human small (duodenum, jejunum, and ileum) and large intestine (ascending, transverse, descending, and sigmoid colon). The c-kit immunoreactive cells in the muscularis externa of the intestinal wall were identified as interstitial cells of Cajal (ICC) and mast cells. ICC were identified by their morphology, localization, and organization based on previous light and electron microscopic studies. In the small intestine, ICC were located primarily in relation to the myenteric plexus of Auerbach, but also in septa between circular muscle lamellae. In the large intestine, ICC were seen in relation to Auerbach’s plexus, but also and in great numbers in the circular muscle layer and in teniae of the longitudinal muscle layer. The morphology of the ICC was similar in the small and large intestine, but the pattern of distribution was obviously different. c-kit immunoreactive mast cells were found predominantly in the inner part of the circular muscle layer. The anti-c-kit method is found to be an easy and reliable method to study at least most of the interstitial cells of Cajal and thereby contribute to further normal and pathological studies. Accepted: 31 July 1997