Development of an array of ion-selective microelectrodes aimed for the monitoring of extracellular ionic activities

In this study, we present the development and the characterization of a generic platform for cell culture able to monitor extracellular ionic activities (K+, NH4+) for real-time monitoring of cell-based responses, such as necrosis, apoptosis, or differentiation. The platform for cell culture is equipped with an array of 16 silicon nitride micropipet-based ion-selective microelectrodes with a diameter of either 2 or 6 microm. This array is located at the bottom of a 200-microm-wide and 350-microm-deep microwell where the cells are cultured. The characterization of the ion-selective microelectrode arrays in different standard and physiological solutions is presented. Near-Nernstian slopes were obtained for potassium- (58.6 +/- 0.8 mV/pK, n = 15) and ammonium-selective microelectrodes (59.4 +/- 3.9 mV/pNH4, n = 13). The calibration curves were highly reproducible and showed an average drift of 4.4 +/- 2.3 mV/h (n = 10). Long-term behavior and response after immersion in physiological solutions are also presented. The lifetime of the sensors was found to be extremely long with a high recovery rate.

Unprecedented Trapping of Difluorooctamolybdate Anions within an α-Polonium Type Coordination Network

New fluorinated hybrid solids [Mo2F2O5(tr2pr)] (1), [Co3(tr2pr)2(MoO4)2F2]·7H2O (2), and [Co3(H2O)2(tr2pr)3(Mo8O26F2)]·3H2O (3) (tr2pr = 1,3-bis(1,2,4-triazol-4-yl)propane) were prepared from the reaction systems consisting of Co(OAc)2/CoF2 and MoO3/(NH4)6Mo7O24, as CoII and MoVI sources, in water (2) or in aqueous HF (1, 3) employing mild hydrothermal conditions. The tr2pr ligand serves as a conformationally flexible tetradentate donor. In complex 1, the octahedrally coordinated Mo atoms are linked in the discrete corner-sharing {Mo2(μ2-O)F2O4N4} unit in which a pair of tr-heterocycles (tr = 1,2,4-triazole) is arranged in cis-positions opposite to “molybdenyl” oxygen atoms. The anti−anti conformation type of tr2pr facilitates the tight zigzag chain packing motif. The crystal structure of the mixed-anion complex salt 2 consists of trinuclear [Co3(μ3-MoO4)2(μ2-F)2] units self-assembling in CoII-undulating chains (Co···Co 3.0709(15) and 3.3596(7) Å), which are cross-linked by tr2pr in layers. In 3, containing condensed oxyfluoromolybdate species, linear centrosymmetric [Co3(μ2-tr)6]6+ SBUs are organized at distances of 10.72−12.45 Å in an α-Po-like network using bitopic tr-linkers. The octahedral {N6} and {N3O3} environments of the central and peripheral cobalt atoms, respectively, are filled by triazole N atoms, water molecules, and coordinating [Mo8O26F2]6− anions. Acting as a tetradentate O-donor, each difluorooctamolybdate anion anchors four [Co3(μ2-tr)6]6+ units through their peripheral Co-sites, which consequently leads to a novel type of a two-nodal 4,10-c net with the Schläfli symbol {32.43.5}{34.420.516.65}. The 2D and 3D coordination networks of 2 and 3, respectively, are characterized by significant overall antiferromagnetic exchange interactions (J/k) between the CoII spin centers on the order of −8 and −4 K. The [Mo8O26F2]6− anion is investigated in detail by quantum chemical calculations.

The nitrogen cycle of tropical montane forest in Ecuador turns inorganic under environmental change

Water-bound nitrogen (N) cycling in temperate terrestrial ecosystems of the Northern Hemisphere is today mainly inorganic because of anthropogenic release of reactive N to the environment. In little-industrialized and remote areas, in contrast, a larger part of N cycling occurs as dissolved organic N (DON). In a north Andean tropical montane forest in Ecuador, the N cycle changed markedly during 1998–2010 along with increasing N deposition and reduced soil moisture. The DON concentrations and the fractional contribution of DON to total N significantly decreased in rainfall, throughfall, and soil solutions. This inorganic turn of the N cycle was most pronounced in rainfall and became weaker along the flow path of water through the system until it disappeared in stream water. Decreasing organic contributions to N cycling were caused not only by increasing inorganic N input but also by reduced DON production and/or enhanced DON decomposition. Accelerated DON decomposition might be attributable to less waterlogging and higher nutrient availability. Significantly increasing NO3-N concentrations and NO3-N/NH4-N concentration ratios in throughfall and litter leachate below the thick organic layers indicated increasing nitrification. In mineral soil solutions, in contrast, NH4-N concentrations increased and NO3-N/NH4-N concentration ratios decreased significantly, suggesting increasing net ammonification. Our results demonstrate that the remote tropical montane forests on the rim of the Amazon basin experienced a pronounced change of the N cycle in only one decade. This change likely parallels a similar change which followed industrialization in the temperate zone of the Northern Hemisphere more than a century ago.

Twentieth Century Increase of Atmospheric Ammonia Recorded in Mount Everest Ice Core

An NH4+ record covering the period A.D. 1845-1997 was reconstructed using an 80.4 m ice core from East Rongbuk Glacier at an elevation of 6450 m on the northern slope of Mount Everest. Variations in NH4+ are characterized by a dramatic increase since the 1950s. The highest NH4+ concentrations occur in the 1980s. They are about twofold more than those in the first half of twentieth century. Empirical orthogonal function (EOF) analysis on the eight major ion (Na+,K+,Mg2+,NH4+,Ca2+,NO3-,SO42- and Cl-) series from this core indicates that NH4+ is loaded mainly on EOF3 (60% of NH4+ variance), suggesting that NH4+ has a unique signature. Instrumental sea level pressure (SLP) and regional temperatures are used to explore the relationship between NH4+ variations and both atmospheric circulation and natural source strength over Asia. Higher NH4+ concentrations are associated with an enhanced winter Mongolian High and a deepened summer Mongolian Low. A positive relationship also exists between NH4+ concentrations and regional temperature changes of the GIS Box 36 (Indian subcontinent), indicating that an increase in temperature may contribute to the strengthening of natural ammonia emissions (e. g., from plants and soils). A close positive correlation between NH4+ and acidic species (SO42- plus NO3-) concentrations suggests that a portion of the increase in NH4+ concentrations could be contributed by enhanced atmospheric acidification. Anthropogenic ammonia emissions from enhanced agricultural activities and energy consumption over Asia in concert with population increase since the 1950s appear also to be a significant factor in the dramatic increase of NH4+ concentrations during the last few decades.

Preliminary Results from the Chemical Records of an 80.4 M Ice Core Recovered from East Rongbuk Glacier, Qomolangma (Mount Everest), Himalaya

High-resolution chemical records from an 80.4 m ice core from the central Himalaya demonstrate climatic and environmental changes since 1844. The chronological net accumulation series shows a sharp decrease from the mid-1950s, which is coincident with the widely observed glacier retreat. A negative correlation is found between the ice-core delta(18)O record and the monsoon precipitation for Indian region 7. The temporal variation of the terrestrial ions (Ca2+ and Mg2+) is controlled by both the monsoon precipitation for Indian regions 3,7 and 8, located directly south and west of the Himalaya, and the dust-storm duration and frequency in the northern arid regions, such as the Taklimakan desert, China. The NH4+ profile is fairly flat until the 1940s, then substantially increases until the end of the 1980s, with a slight decrease during the 1990s which may reflect new agricultural practices. The SO42- and NO3- profiles show an apparent increasing trend, especially during the period 1940s-80s. Moreover, SO42- concentrations for the East Rongbuk Glacier core are roughly double that of the nearby Dasuopu core at Xixabangma, Himalaya, due to local human activity including that of climbing teams who use gasoline for cooking, energy and transport.

Springtime Nutrient and Phytoplankton Dynamics on Georges Bank

The dynamics of phytoplankton and nutrients before, during and after the winter-spring bloom on Georges Bank were studied on 6 monthly survey cruises from January to June 1999. We measured hydrography, phytoplankton cell densities, chlorophyll a, dissolved inorganic nutrients (NO3 + NO2, NH4, Si(OH)(4), PO4), dissolved organic nitrogen (DON) and phosphorus (DOP), particulate organic carbon (POC) and nitrogen (PON) and total particulate phosphorus (TPP). We present evidence that phytoplankton production may be significant year-round, and that the winter-spring bloom may have started in January. From January to April the phytoplankton was comprised almost exclusively of diatoms, reaching cell densities in March and April of ca. 450 cells ml(-1); chlorophyll a concentrations exceeded 10 mug l(-1) in April. Diatoms decreased to relatively low levels in May (< 50 x 10(3) cells l(-1)) and increased again in June (>300 x 10(3) cells l(-1)). Densities of dinoflagellates and nanoflagellates were low (< 10 x 10(3) cells l(-1)) from January to April, and increased in May and June to nearly 300 x 10(3) cells l(-1). Nitrate + nitrite concentrations in January were <3 muM in the shallow, central portion of the bank and decreased steadily each month. Silicate was also <3 muM over an even larger area of the central bank in January and declined to <1.5 muM over most of the Bank in April. The data suggest that silicate depletion, not DIN, contributed to the cessation of the diatom bloom. Regeneration of silicate occurred in May and June, presumably as a result of rising water temperatures in late spring which increased the dissolution rate of diatom frustules from the earlier diatom bloom. Dissolved organic nitrogen may have been utilized at the start of the winter-spring bloom; concentrations were ca, 14 muM in January, dropping to < 6 mug l(-1) in February, after which DON concentrations steadily rose to > 15 mug l(-1) in June. Overall micro-and nanoplankton biomass, measured as POC, PON and TPP, increased over the 6 mo period, as did nutritional quality of that biomass as indicated by declining C:N ratios. Our results suggest there may have been an increase in the heterotrophic component of the plankton in May and June which coincided with a second burst in diatom abundance. We discuss general features of planktonic production and nutrient dynamics with respect to year-round production on the Bank.

Does fog chemistry in Switzerland change with altitude?

During two extended summer seasons in 2006 and 2007 we operated two battery driven versions of the Caltech active strand cloud water collector (MiniCASCC) at the Niesen mountain (2362 m a.s.l.) in the northern part of the Swiss Alps, and two devices at the Lägeren research tower (690 m a.s.l.) at the northern boundary of the Swiss Plateau. During these two field operation phases we gained weekly samples of fog water, where we analyzed the major anions and cations, and the isotope ratios of fog water (in form of δ2H and δ18O). Dominant ions in fog water at all sites were NH4+, NO3−, and SO42 −. Compared to precipitation, the enrichment factors in fog water were in the range 5–9 at the highest site, Niesen Kulm. We found considerably lower summertime ion loadings in fog water at the two Alpine sites than at lower elevations above the Swiss Plateau. The lowest ion concentrations were found at the Niesen Kulm site at 2300 m a.s.l., whereas the highest concentrations (a factor 7 compared to Niesen Kulm) were found in fog water at the Lägeren site. Occult nitrogen deposition was estimated from fog frequency and typical fog water flux rates. This pathway contributes 0.3–3.9 kg N ha− 1 yr− 1 to the total N deposition at the highest site on Niesen mountain, and 0.1–2.2 kg N ha− 1 yr− 1 at the lower site. These inputs are the reverse of ion concentrations measured in fog due to the 2.5 times higher frequency of fog occurrence at the mountain top (overall fog occurrence was 25% of the time) as compared to the lower Niesen Schwandegg site. Although fog water concentrations were on the lower range reported in earlier studies, fog water is likely to be an important N source for Northern Alpine ecosystems and might reach values up to 16% of the total N deposition and up to 75% of wet N deposition by precipitation.

Relationships between functional traits and inorganic nitrogen acquisition among eight contrasting European grass species

Backgrounds and Aims Leaf functional traits have been used as a basis to categoize plants across a range of resource-use specialization, from those that conserve available resources to those that exploit them. However, the extent to which the leaf functional traits used to define the resource-use strategies are related to root traits and are good indicators of the ability of the roots to take up nitrogen (N) are poorly known. This is an important question because interspecific differences in N uptake have been proposed as one mechanism by which species coexistence may be determined. This study therefore investigated the relationships between functional traits and N uptake ability for grass species across a range of conservative to exploitative resource-use strategies.Methods Root uptake of NH4+ and NO3-, and leaf and root functional traits were measured for eight grass species sampled at three grassland sites across Europe, in France, Austria and the UK. Species were grown in hydroponics to determine functional traits and kinetic uptake parameters (Imax and Km) under standardized conditions.Key Results Species with high specific leaf area (SLA) and shoot N content, and low leaf and root dry matter content (LDMC and RDMC, respectively), which are traits associated with the exploitative syndrome, had higher uptake and affinity for both N forms. No trade-off was observed in uptake between the two forms of N, and all species expressed a higher preference for NH4+.Conclusions The results support the use of leaf traits, and especially SLA and LDMC, as indicators of the N uptake ability across a broad range of grass species. The difficulties associated with assessing root properties are also highlighted, as root traits were only weakly correlated with leaf traits, and only RDMC and, to a lesser extent, root N content were related to leaf traits.

Biochemical changes in barley plants after excessive supply of copper and manganese

The present study was undertaken to identify changes in some important proteins involved in CO2 fixation (Rubisco, Rubisco activase (RA), Rubisco binding protein (RBP)), NH4+ assimilation (glutamine synthetase (GS) and glutamate synthase (GOGAT)), using immunoblotting, and in the antioxidative defense as a result of Cu or Mn excess in barley leaves (Hordeum vulgare L. cv. Obzor). Activities and isoenzyme patterns of superoxide dismutase (SOD), ascorbate peroxidase (APX), guaiacol peroxidase (GPX) and catalase (CAT), as well as the levels of ascorbate (ASC), non-protein sulfhydryl groups, hydrogen peroxide and oxidative damage to proteins were determined. Data were correlated to the accumulation of Cu or Mn in the leaves after 5 days supply of heavy metal (HM) excess in the nutrient solution. In the highest Cu excess (1500 μM), Rubisco LS and SS were reduced considerably whereas under the highest Mn concentrations (18,300 μM) only minor changes in Rubisco subunits were detected. The RBP was diminished under the highest concentrations of both Cu or Mn. The bands of RA changed differently comparing Cu and Mn toxicity. GS decreased and GOGAT was absent under the highest concentration of Cu. At Mn excess Fd-GOGAT diminished whereas GS was not apparently changed. The development of toxicity symptoms corresponded to an accumulation of Cu or Mn in the leaves and to a gradual increase in protein carbonylation, a lower SOD activity and elevated CAT and GPX activities. APX activity was diminished under Mn toxicity and was not changed under Cu excess. Generally, changes in the isoenzyme profiles were similar under both toxicities. An accumulation of H2O2 was observed only at Mn excess. Contrasting changes in the low-molecular antioxidants were detected when comparing both toxicities. Cu excess affected mainly the non-protein SH groups, while Mn influenced the ASC content. Oxidative stress under Cu or Mn toxicity was most probably the consequence of depletion in low-molecular antioxidants as a result of their involvement in detoxification processes and disbalance in antioxidative enzymes. The link between heavy metal accumulation in leaves, leading to different display of oxidative stress, and changes in individual chloroplast proteins is discussed in the article.

N budget and NH3 exchange of a grass/clover crop at two levels of N application

Atmospheric ammonia (NH3) exchange during a single growing season was measured over two grass/clover fields managed by cutting and treated with different rates of mineral nitrogen (N) fertilizer. The aim was to quantify the total NH3 exchange of the two systems in relation to their N budget, the latter was split into N derived from symbiotic fixation, from fertilization, and from the soil. The experimental site was located in an intensively managed agricultural area on the Swiss plateau. Two adjacent fields with mixtures of perennial ryegrass (Lolium perenne L.), cocks foot (Dactylis glomerata L.), white clover (Trifolium repens L.) and red clover (Trifolium pratense L.) were used. These were treated with either 80 or 160 kg N ha−1 applied as NH4NO3 fertilizer in equal portions after each of four cuts. Continuous NH3 flux measurements were carried out by micrometeorological techniques. To determine the contribution of each species to the overall NH3 canopy compensation point, stomatal NH3 compensation points of the individual plant species were determined on the basis of NH4+ + NH3 (NHx) concentrations and pH in the apoplast. Symbiotic N2 fixation was measured by the 15N dilution method.

Regulation of Sulfate Assimilation by Nitrogen in Arabidopsis

Using Arabidopsis, we analyzed the effect of omission of a nitrogen source and of the addition of different nitrogen-containing compounds on the extractable activity and the enzyme and mRNA accumulation of adenosine 5′-phosphosulfate reductase (APR). During 72 h without a nitrogen source, the APR activity decreased to 70% and 50% of controls in leaves and roots, respectively, while cysteine (Cys) and glutathione contents were not affected. Northern and western analysis revealed that the decrease of APR activity was correlated with decreased mRNA and enzyme levels. The reduced APR activity in roots could be fully restored within 24 h by the addition of 4 mM each of NO3 −, NH4 +, or glutamine (Gln), or 1 mM O-acetylserine (OAS). 35SO4 2− feeding showed that after addition of NH4 +, Gln, or OAS to nitrogen-starved plants, incorporation of 35S into proteins significantly increased in roots; however, glutathione and Cys labeling was higher only with Gln and OAS or with OAS alone, respectively. OAS strongly increased mRNA levels of all three APR isoforms in roots and also those of sulfite reductase, Cys synthase, and serine acetyltransferase. Our data demonstrate that sulfate reduction is regulated by nitrogen nutrition at the transcriptional level and that OAS plays a major role in this regulation.

Oceanography during TYDEMAN cruise DCM (DeepChlorMax) to the Equatorial Atlantic

The cruise with RV Tydeman was devoted to study permanently stratified plankton systems in the (sub)tropical ocean, which are characterised by a deep chlorophyll peak between 80 and 150 m. To minimise lateral effects by horizontal transport of nutrients and organic matter from river outflow and upwelling regions, stations were selected in the middle of the North Atlantic Ocean between the continents of America and Africa. (5 - 35° N and 50 - 15° W). Here the vertical distributions of light and nutrients control the abundance and growth of autotrophic algae in the thermically stratified water column. This phytoplankton is numerically dominated by the prokaryotic picoplankters Synechococcus spp. and Prochlorococcus spp., which are smaller than 2 ?m. The productivity of the 100 to 150 m deep euphotic zone can be high, because a high heterotrophic/autotrophic biomass ratio induces a rapid regeneration of nutrients and inorganic carbon. Primary grazers are mainly micro-organisms such as heterotrophic nannoflagellates and ciliates, which feed on the small algae and on bacteria. Heterotrophic bacteria can outnumber the autotrophic algae, because their number is related to the substrate pools of dissolved and particulate dead organic matter. These DOC and detritus pools reach equilibrium at a concentration, where the rate of their production (proportional to algal biomass) equals their mineralisation and sinking rate (proportional to the concentration and weight of POC and detritus). At a relatively low value of the weight-specific loss rates, the equilibrium concentration of these carbon pools and their load of bacteria can be high. The bacterial productivity is proportional to the mineralisation rate, which in a steady state can never be higher than the rate of primary production. Hence the ratio in turnover rate of bacteria and autotrophs tends to be reciprocally proportional to their biomass ratio.