The interaction of chloride with the sulfate transport system of ehrlich ascites tumor cells

The effect of Cl^? on SO₄^?2 efflux was studied in both Cl^?-containing and Cl^?-free ascites tumor cells loaded with ³⁵SO₄^?2 to test the hypothesis that Cl^?-SO₄^?2 exchange is mediated by the same mechanism responsible for SO₄^?2-self exchange. The addition of Cl^?-free, ³⁵SO₄^?2 loaded cells to a SO₄^?2-free, Cl^? medium results in: (1) SO₄^?2 efflux that is dependent on the extracellular Cl^? concentration (Km = 4.85 mM; k_e = 0.048 min^?1 at 50 mM Cl^?) and (2) net Cl^?-uptake that exceeds SO₄^?2 loss. Both SITS (4-acetamido-4′-isothiocyanostilbene-2,2′-disulfonate) and ANS (1-anilino-8-napthalene sulfonate) inhibit SO₄^?2 efflux but are without effect on Cl^? uptake. The addition of Cl^?-containing, ³⁵SO₄^?2 loaded cells to a SO₄^?2-free, C1^? medium results in: (1) a slight gain in cellular Cl^? and (2) k efor SO₄^?2 efflux identical to that for Cl^?-free cells. The results are compatible with the suggestion that: (1) Cl^? interacts with a membrane component responsible for transmembrane SO₄^?2 movement; (2) Cl^? interaction stimulates the rate of unidirectional SO₄^?2 efflux from cells initially free of Cl^? as well as the rate of SO₄^?2 turnover in cells maintained in the steady state with respect to Cl^? and SO₄^?2; and (3) in the case of cells initially free of Cl^?, the Cl^?-SO₄^?2 pathway represents only a small fraction of the total unidirectional Cl^?-influx the remainder being compatible with the electroneutral accumulation of NaCl and KCl.     

Further observations on metabolic responses in hamster cells: changes in UDPG dehydrogenase activity.

The effects of phloretin, H₂DIDS (4,4′-diisothiocyano-1,2-diphenylethane-2,2′-disulfonate) and SO₄^?2 on anion transport in Ehrlich ascites tumor cells was studied in an effort to determine whether Cl^? and SO₄^?2 share a common transport mechanism. Sulfate, in the presence of constant extracellular Cl^? (100 mM), reduces Cl^? self-exchange by 43% (40 mM SO₄^?2) and Cl^??SO₄^?2 exchange by 36% (25 mM Cl^?/O SO₄^?2) compared to 25 mM Cl^?/50 mM SO₄^?2. Phloretin blocks without delay and to the same extent the self-exchange of both Cl^? and SO₄^?2. For example, at 10^?4 M phloretin, anion transport is inhibited 28% which increases to 78% at 5 × 10^?4 M. Reversibly bound H₂DIDS also inhibits the self-exchange of both Cl^? and SO₄^?2. However, at all H₂DIDS concentrations tested (0.5 ? 10 × 10^?5 M) SO₄^?2 transport was far more susceptible to inhibition than that of Cl^?. H₂DIDS when irreversibly bound to the cell inhibits SO₄^?2 but not Cl^? transport The results of these experiments are consistent with the postulation that both Cl^? and SO₄^?2 are transported by a common mechanism possessing two reactive sites.     

Uptake and loss of nitrite from the blood of rainbow trout, Salmo gairdneri Richardson, and Atlantic salmon, Salmo salar L. in fresh water and in dilute sea water

The acute toxicity of nitrite (NO^?₂) to salmonids is strongly ameliorated by chloride (Cl^?) ions rendering it almost harmless in most fresh waters apart from those with low Cl^? content. In Cl^? poor fresh water external NO^?₂ is concentrated in the blood plasma until it is at approximately the same molar concentration as haemoglobin (about 8 mmol) and at this point most of the haemoglobin has been oxidized to methaemoglobin this being a contributory cause of death. Two theories are advanced to account for NO^?₂ concentration in the blood. The first supposes that gills are impermeable to NO^?₂ but allow its conjugate acid nitrous acid (HNO₂) to diffuse into the blood where it dissociates according to the blood pH value. Thus NO^?₂ will accumulate in the blood plasma if it has a higher pH value than the water. The second supposes that the Cl^? uptake mechanism in the freshwater gill has an affinity for NO^?₂ and accounts for the fact that NO^?₂ entry to the blood is suppressed when external Cl^? is present in significant amounts. The results also suggest that NO^?₂ and Cl^? behave similarly as diffusing ions. Thus NO^?₂ diffusion into the blood of seawater fish and from the blood of NO^?₂ loaded freshwater fish occurs at approximately the same rate as the corresponding Cl^? fluxes. Nitrite loss from seawater fish is thought to be mainly by diffusion although there is some evidence for the active Cl^? extrusion mechanism having a weak affinity for nitrite.     

Effect of Light Intensity on Growth, CO2 Fixation, and Chlorophyll and Polar Lipid Production in Germinating Polytrichum commune Spores

The dry matter production in Polytrichum commune protonemata was increased when the light intensity was increased from 0 to 160 μE m^?2 s^?1, and at 160 μE m^?2 s^?1 production was about 200% of that found at 17 μE m^?2 s^?1. Production of chlorophyll (Chl) was increased by increasing light intensity from 0 to 17 μE m^?2 s^?1, but decreasing at light intensities above 17 μE m^?2 s^?1. At 160 μE m^?2 s^?1 the production of Chl was only about 50% of that at 17 μE m^?2 s^?1. The rate of CO₂ fixation was low (0.31 μg CO₂/mg Chi × h) at the light intensity of 17 μE m^?2 s^?1 as compared with that at 160 μE m^?2 s^?1 (0.83 μg CO₂/mg Chi × h). Production of mono- (MGDG) and diglycosyl diglycerides (DGDG) was closely associated with that of chlorophylls. At the higher light intensity (160 μE m^?2 s^?1) production of glycolipids was about 60% of that at 17 μE m^?2 s^?1. Production of more polar lipids was less affected by light intensity. Light intensity also affected the fatty acid pattern of the lipid fractions. The effect was most pronounced in the MGDG fraction, where the proportion of C 18: 3ω3 + C 16: 3ω3 was higher at the higher light intensity.     

Measurement of net ecosystem exchange,productivity and respiration in three spruce forests in Sweden shows unexpectedly large soil carbon losses

Measurement of net ecosystem exchange was made using the eddy covariance method above three forests along a north-south climatic gradient in Sweden: Flakaliden in the north, Knottåsen in central and Asa in south Sweden. Data were obtained for 2 years at Flakaliden and Knottåsen and for one year at Asa. The net fluxes (N_ep) were separated into their main components, total ecosystem respiration (R_t) and gross primary productivity (P_g). The maximum half-hourly net uptake during the heart of the growing season was highest in the southernmost site with ?0.787 mg CO₂ m^?2 s^?1 followed by Knottåsen with ?0.631 mg CO₂ m^?2 s^?1 and Flakaliden with ?0.429 mg CO₂ m^?2 s^?1. The maximum respiration rates during the summer were highest in Knottåsen with 0.245 mg CO₂ m^?2 s^?1 while it was similar at the two other sites with 0.183 mg CO₂ m^?2 s^?1. The annual N_ep ranged between uptake of ?304 g C m^?2 year^?1 (Asa) and emission of 84 g C m^?2 year^?1 (Knottåsen). The annual R_t and P_g ranged between 793 to 1253 g C m^?2 year^?1 and ?875 to ?1317 g C m^?2 year^?1, respectively. Biomass increment measurements in the footprint area of the towers in combination with the measured net ecosystem productivity were used to estimate the changes in soil carbon and it was found that the soils were losing on average 96–125 g C m^?2 year^?1. The most plausible explanation for these losses was that the studied years were much warmer than normal causing larger respiratory losses. The comparison of net primary productivity and P_g showed that ca 60% of P_g was utilized for autotrophic respiration.     

Thymic lymphocytes: II. Phenotypic modifications of thymocytes after concanavalin A stimulation in the presence of interleukin 2: Early modifications of Lyt 1+2+ subset and later proliferation of cells with more mature phenotypes

Cortical thymocytes are devoid of any immune function, as tested by presently available techniques. The ability of this subpopulation to respond to mitogens or antigens in the presence of interleukin 2 (IL-2) produced by activated mature T lymphocytes has been claimed but is still questioned. In an attempt to study the participation of the different thymocyte subsets and especially that of the cortical type, phenotypic modifications were examined during concanavalin A activation in the presence of IL-2. An immunofluorescent double labeling technique with anti-Lyt 1 and anti-Lyt 2 antibodies was used which led to the determination of four different phenotypes: Lyt 1⁺2⁺, Lyt 1⁺2^?, Lyt 1^?2⁺, and Lyt 1^?2^?. Careful analysis of cell viability in culture and expression of the results in absolute numbers of living cells per culture allowed us to follow modifications of small cellular subsets. Cultures of total thymocytes and PNA-agglutinated (enriched in Lyt 1⁺2⁺ cells) and non-PNA-agglutinated cells (enriched in Lyt 1⁺2^?, Lyt 1^?2⁺, and Lyt 1^?2^? cells) were studied. It was shown that thymocyte activation began by early phenotypic modifications which took place within the first 2 hr of culture but only when Con A plus IL-2 were used. These modifications imply the reduction of the Lyt 1⁺2⁺ pool and a compensatory enhancement of Lyt 1^?2⁺ and Lyt 1^?2^? cells, without modification of the total cell number or [³H]thymidine incorporation. These early phenotypic changes are interpreted as the modulation of antigens on the surface of Lyt 1⁺2⁺ cells. The second phase of thymocyte activation implies cell death (essentially Lyt 1⁺2⁺ cells) and cell proliferation. The cells which specifically proliferate in the presence of Con A and IL-2 are Lyt 1⁺2^? and Lyt 1^?2⁺, the latter always being present in greater number. Cell survival and absolute number of Lyt 1⁺2^? and Lyt 1^?2⁺ cells in the activated PNA^?-enriched population are always higher than in total thymocyte and PNA⁺ cells cultures. Thus, if Lyt 1⁺2⁺ cortical thymocytes do not proliferate by themselves, they seem to intervene by providing Lyt 1^?2⁺ cells which proliferate secondarily.     

Seasonal changes in the chlorophyll content and quantum efficiency of the moss Brachythecium rutabulum

Abstract

The concentration of chlorophyll a, b, and total chlorophyll have been monitored on a seasonal basis in Brachythecium rutabulum. Total chlorophyll increases during summer full canopy conditions from 1.70 mg chl g^?1 on 8 May to 11.1 mg chl g^?1 on 11 October. Photosynthetic-illumination curves show that during this period light saturation declines from 200 μmol m^?2s^?1 to 30 μmol m^?2s^?1 by 6 July, and light compensation falls dramatically from 65 μmol m^?2s^?1 to 4 μmol m^?2s^?1. The data also appear to support the conclusion that there is concurrently an increase in the density of photosynthetic units by the end of September.     

Kinetics of chloride transport in the cyanobacterium Synechococcus R-2 (Anacystis nidulans, S. leopoliensis) PCC 7942

The kinetics of the light-driven Cl^? uptake pump of Synechococcus R-2 (PCC 7942) were investigated. The kinetics of Cl^? uptake were measured in BG-11 medium (pH_o, 7·5; [K⁺]_o, 0·35 mol m^?3; [Na⁺]_o, 18 mol m^?3; [Cl^?]_o, 0·508 mol m^?3) or modified media based on the above. Net³⁶Cl^? fluxes (?Cl^?_o,i) followed Michaelis-Menten kinetics and were stimulated by Na⁺ [18 mol m^?3 Na⁺ BG-11 ?Cl^?_max= 3·29±0·60 (49) nmol m^?2 s^?1 versus Na⁺-free BG-11 ?Cl^?_max= 1·02±0·13 (54) nmol m^?2 s^?1] but the Km was not significantly different in the presence or absence of Na⁺ at pH_o 10; the Km was lower, but not affected by the presence or absence of Na⁺ [Km = 22·3±3·54 (20) mmol m^?3]. Na⁺ is a non-competitive activator of net ?Cl^?_o,i. High [K⁺]_o (18 mol m^?3) did not stimulate net ?Cl^?_o,i or change the Km in Na⁺-free medium. High [K⁺]_o (18 mol m^?3) added to Na⁺ BG-11 medium decreased net ?Cl^?_o,i [18 mol m^?3K⁺ BG-11; ?Cl^?_max= 2·50±0·32 (20) nmol m^?2 s^?1 versus BG-11 medium; ?Cl^?_max= 3·35±0·56 (20) nmol m^?2 s^?1] but did not affect the Km 55·8±8·100 (40) mmol m^?3]. Na⁺-stimulation of net ?Cl^?_o,i followed Michaelis-Menten kinetics up to 2–5 mol m^?3 [Na⁺]_o but higher concentrations were inhibitory. The Km for Na⁺-stimulation of net ?Cl^?_o,i [K_1/2(Na⁺)] was different at 47 mmol m^?3 [Cl^?]_o (K_1/2[Na⁺] = 123±27 (37) mmol m^?3]. Li⁺ was only about one-third as effective as Na⁺ in stimulating Cl^? uptake but the activation constant was similar [K_1/2(Li⁺) = 88±46 (16) mmol m^?3]. Br^? was a competitive inhibitor of Cl^? uptake. The inhibition constant (Ki) was not significantly different in the presence and absence of Na⁺. The overall Ki was 297±23 (45) mmol m^?3. The discrimination ratio of Cl^? over Br^? (δCl^?/δBr^?) was 6·38±0·92 (df = 147). Synechococcus has a single Na⁺-stimulated Cl^? pump because the Km of the Cl^? transporter and its discrimination between Cl^? and Br^? are not significantly different in the presence and absence of Na⁺. The Cl^? pump is probably driven by ATP.     

Experimental determination of the redox potential of the superoxide radical O 2

The redox potential of ^?O₂^? was determined based on the dependence of the electron transfer reaction from ^?O₂^? upon the known redox potential of various acceptors A (including a range of quinones, dyes and ferricyanide). The efficiencies and the rates of these electron transfer processes were determined, using the technique of pulse radiolysis, by monitoring the formation kinetics of the semiquinone radical anions at the appropriate wavelength. From the percentage efficiency versus E^o′ plot, an E^o′ value of + 0.15 ± 0.01 V at pH 7.0 and 22°C, or E^o = + 0.57 ± 0.01 V, for the ${}^{\mathrm{?}}\text{O}{}_2{}^{\mathrm{?}}\text{O}{}_2$ couple was obtained. The rate k(^?O₂^? + A → O₂ + ^?A^?) = 9.8 × 10⁸ M^?1 sec^?1 where A = p-benzoquinone. The E^o value for the ^?HO₂ radical is > 1.0 V. It was also found that hydroquinone can quantitatively reduce ^?O₂^? to H₂O₂, k(^?O₂^? + QH₂ → QH^? + HO₂^?) = 1.6 ± 0.1 × 10⁷ M^?1 sec^?1 at pH 7.0 and 22°C.     

Ecosystem–atmosphere exchange of CH4 and N2O and ecosystem respiration in wetlands in the Sanjiang Plain, Northeastern China

Natural wetlands are critically important to global change because of their role in modulating atmospheric concentrations of CO₂, CH₄, and N₂O. One 4‐year continuous observation was conducted to examine the exchanges of CH₄ and N₂O between three wetland ecosystems and the atmosphere as well as the ecosystem respiration in the Sanjiang Plain in Northeastern China. From 2002 to 2005, the mean annual budgets of CH₄ and N₂O, and ecosystem respiration were 39.40 ± 6.99 g C m^?2 yr^?1, 0.124 ± 0.05 g N m^?2 yr^?1, and 513.55 ± 8.58 g C m^?2 yr^?1 for permanently inundated wetland; 4.36 ± 1.79 g C m^?2 yr^?1, 0.11 ± 0.12 g N m^?2 yr^?1, and 880.50 ± 71.72 g C m^?2 yr^?1 for seasonally inundated wetland; and 0.21 ± 0.1 g C m^?2 yr^?1, 0.28 ± 0.11 g N m^?2 yr^?1, and 1212.83 ± 191.98 g C m^?2 yr^?1 for shrub swamp. The substantial interannual variation of gas fluxes was due to the significant climatic variability which underscores the importance of long‐term continuous observations. The apparent seasonal pattern of gas emissions associated with a significant relationship of gas fluxes to air temperature implied the potential effect of global warming on greenhouse gas emissions from natural wetlands. The budgets of CH₄ and N₂O fluxes and ecosystem respiration were highly variable among three wetland types, which suggest the uncertainties in previous studies in which all kinds of natural wetlands were treated as one or two functional types. New classification of global natural wetlands in more detailed level is highly expected.     

Bacterial production in the carbon flow of a central European stream, the Breitenbach

1. Over the last 30 years, many investigations have been performed on the dynamics of bacteria and organic matter in the Breitenbach, a first‐order stream in central Germany. The data now available allow a synthesis of the role of bacteria in the carbon budget, as an example of the general importance of bacteria in stream ecosystems. 2. Comparing measured and estimated inputs and outputs to the ecosystem, the organic matter budget of the Breitenbach is fairly balanced: 1.84 kg C m^?2 year^?1 (sum of inputs) versus 1.88 kg C m^?2 year^?1 (sum of outputs). No major missing link remains. 3. The basis of the food web in the Breitenbach is mainly allochthonous organic matter (dissolved and particulate 1.02 and 0.42 kg C m^?2 year^?1, respectively). Autochthonous gross primary production is 0.4 kg C m^?2 year^?1. Most of the organic matter leaves the stream via transport to the River Fulda (dissolved and particulate 0.74 and 0.34 kg C m^?2 year^?1, respectively), the rest by respiration (0.80 kg C m^?2 year^?1 or 43% of total outputs). 4. Bacteria constitute an important part (36%) of heterotrophic biomass (average: 0.004 kg m^?2 bacterial C of 0.011 kg m^?2 total heterotrophic C). Bacteria also account for the major fraction (71%) of heterotrophic production: 0.20 of 0.28 kg C m^?2 year^?1 total heterotrophic production. Bacterial production in the Breitenbach is similar in magnitude to the estimate of photoautotrophic net primary production: both approximately 0.20 kg C m^?2 year^?1. 5. Protozoa, the main consumers of bacteria in the Breitenbach, consume approximately one‐third of bacterial production (0.07 kg C m^?2 year^?1). Small metazoa (meiofauna, <0.5 mm) play a lesser role in the consumption of bacteria, consuming <0.01 kg bacterial C m^?2 year^?1. Larger metazoa (macrofauna, >0.5 mm) consume approximately 10% of bacterial production. Although this is a considerable amount of the carbon resources needed by the macrofauna (0.02 kg C m^?2 year^?1 of bacterial production versus 0.06 kg C m^?2 year^?1 macrofauna production plus respiration), the carbon demand of the macrofaunal community is met to a larger extent by particulate organic matter than by bacteria. 6. Bacteria are the main decomposers in the Breitenbach. They account for 78% of heterotrophic respiration (0.47 of 0.60 kg C m^?2 year^?1) and 59% of total respiration (0.47 of 0.80 kg C m^?2 year^?1).     

Photon irradiance required to support optimal growth and interrelations between irradiance and pigment composition in the green alga Haematococcus pluvialis

The aim of the work was to find the optimal photon irradiance for the growth of green cells of Haematococcus pluvialis and to study the interrelations between changes in photochemical parameters and pigment composition in cells exposed to photon irradiances between 50 and 600?µmol?m^?2?s^?1 and a light:dark cycle of 12:12?h. Productivity of cultures increased with irradiance. However, the rate of increase was higher in the range 50–200?µmol?^?2?s^?1. The carotenoid content increased with increasing irradiance, while the chlorophyll content decreased. The maximum quantum yield of PSII (F_v/F_m) gradually declined from 0.76 at the lowest irradiance of 50?µmol?^?2?s^?1 to 0.66 at 600?µmol?^?2?s^?1. Photosynthetic activity showed a drop at the end of the light period, but recovered fully during the following dark phase. A steep increase in non-photochemical quenching was observed when cultures were grown at irradiances above 200?µmol?^?2?s^?1. A sharp increase in the content of secondary carotenoids also occurred above 200?µmol?m^?2?s^?1. According to our results, with H. pluvialis green cells grown in a 5-cm light path device, 200?µmol?^?2?s^?1 was optimal for growth, and represented a threshold above which important changes in both photochemical parameters and pigment composition occurred.     

CRITICAL IRRADIANCE LEVELS AND THE INTERACTIVE EFFECTS OF QUANTUM IRRADIANCE AND DOSE ON GAMETOGENESIS IN THE GIANT KELP,MACROCYSTIS PYRIFERA1

Gametophytes of Macrocystis pyrifera (L.) C. Ag. were cultured under a series of quantum irradiances in three photoperiod regimes. The quantum irradiances in each photoperiod were adjusted to provide equal daily irradiation dosages between photoperiods which allowed a critical examination of the interactions between quantum irradiance and quantum dose in determining gametophyte fertility. The lowest quantum irradiance which stimulated gametogenesis in more than 50% of the female gametophytes was 5 μE·m^?2·s^?1. The saturating irradiance was ca. 10 μE·m^?2·s^?1 at photoperiods of 12 h or greater. In terms of daily quantum dose, the lowest dose at which greater than 50% gametogenesis occurred was 0.2 E·m^?2·d^?1. However, this critical quantum dose was higher (0.4 E·m^?2·d^?1) when instantaneous irradiances were less than 5 μE·m^?2·s^?1. The saturation quantum dose was also affected by the rate at which the quantum dose was received and varied from 0.4 to 0.8 E·m^?2·d^?1. Gametophytes in all three photoperiods reached 100% fertility at quantum irradiances above 5 μE·m^?2·s^?1. Photoperiod effects were small and could be accounted for by quantum dosage effects.     

CO2 Supersaturation and Net Heterotrophy in a Tropical Estuary (Cochin, India): Influence of Anthropogenic Effect

Carbon biogeochemistry of a tropical ecosystem (The Cochin Estuary, India) undergoing increased human intervention was studied during February (premonsoon), April (early monsoon) and September (monsoon) 2005. The Cochin estuary sustains high levels of pCO₂ (up to 6000 μatm) and CO₂ effluxes (up to 274 mmolC m^?2 d^?1) especially during monsoon. A first-order estimate of the carbon mass balance shows that net production of dissolved inorganic carbon is an order of magnitude higher than the net loss of dissolved and particulate organic carbon from the estuary. This imbalance is attributed to the organic inputs to the estuary through anthropogenic supplies. The bacteria-mediated mineralization of organic matter is mainly responsible for the build-up of pCO₂ and increased CO₂ emission to the atmosphere indicating heterotrophy. The linear correlation between excess CO₂ and apparent oxygen utilization indicates respiration as the chief mechanism for CO₂ supersaturation. An increase in the net negative ecosystem production (–ve NEP) between premonsoon (?136 mmolC m^?2 d^?1 or ?376 MgC d^?1) and monsoon (?541 mmolC m^?2 d^?1 or ?1500 MgC d^?1) is supported by a corresponding increase in O₂ influxes from 17 mmol O₂ m^?2 d^?1 (126 MgC d^?1) to ?128 mmol O₂ m^?2 d^?1 (?946 MgC d^?1) and CO₂ emissions from 65 mmolC m^?2 d^?1 (180 MgC d^?1) to 267 mmolC m^?2 d^?1 (740 MgC d^?1). There is a significant north-south gradient in metabolic rates and CO₂ fluxes attributable to the varying flow patterns and anthropogenic inputs into the estuary. The study reveals that the Cochin estuary, a previously autotrophic (CO₂ sink) system, has been transformed to a heterotrophic (CO₂ source) system following rapid urbanization and industrialization. Moreover, the export fluxes from the Cochin estuary appear to be quite important in sustaining net heterotrophy in the southeastern Arabian Sea.     

Influx and efflux of inorganic carbon in Synechococcus UTEX625

The CO₂ and HCO₃^? fluxes in air-grown cells of Synechococcus UTEX 625 al pH 8-0 were measured during dark to light and light to dark transitions using a mass spectrometer and sampling of the reaction medium. The kinetic parameters for initial uptake of CO₂ and HCO₃^? were determined during the initial period of illumination. The development of the internal C_i pool was followed up to steady-state photosynthesis, which occurred when the size of the internal inorganic carbon pool remained apparently constant for a limited period. The experimental procedure confirmed that only CO₂ transport occurred with 100mmolm^?3 Na⁺ and that both CO₂ and HCO^?3 transport occurred with 25molm^?3 Na⁺. The K_1/2 values of initial CO₂ and HCO₃ uptake were 0.7 and 17.2 mmolm^?3respectively and agreed closely with the K_1/2 values of net CO₂ and HCO₃^? transport during steady-state photosynthesis, which were 0.66 and 17.1 mmolm^?3 respectively. Maximum rates of CO₂and HCO₃^? transport were 423 and 219mmolh^?1 g^?1 Chl. Maximum CO₂ efflux observed upon darkening was 118mmolh^?1 g^?1 Chl. A permeability coefficient of the cell for CO₂ of 3 × 10^?8 m s^?1 was determined from the dark CO₂ efflux assuming an internal pH of 7.2 in the dark. Following the initial CO₂ uptake in the light, the extracellular [CO₂] steadily declined when only CO₂ transport was allowed, but an increase in the extracellular [CO₂] when HCO₃^? transport was allowed to proceed suggested that an enhanced CO₂ efflux occurred as a result of the larger size of the intracellular C_i pool.     

The presence of discrete receptors for prostaglandin F2 alpha in the cell membranes of bovine corpora lutea.

The cell membranes isolated from bovine corpora lutea bound ³H-prostaglandin (PG) F₂α with high affinity and specificity. The specific binding of ³H-PGF₂α was detectable at 10^?10M added ³H-PGF₂α and reached saturation at 10^?7M to 10^?6M. Unlabeled PGF₂α, as low as 10^?9M, inhibited the binding of ³H-PGF₂α with complete inhibition occurring at 10^?6M. The Scatchard analysis of equilibrium binding data revealed that the PGF₂α receptors are heterogeneous: Kd₁?5.1 × 10^?9M, n?289 fmoles/mg protein; Kd₂?1.8 × 10^?8M, n?780 fmoles/mg protein. The relative affinities of various other PGs for binding to PGF₂α receptors were (PGF₂α?100%): PGF₁α?17.5; PGE₁?0.8; PGE₂?22.4; PGA₁?0.007; PGB₁?0.01. The specificity and affinity of ³H-PGF₂α binding is consistent with the possibility that this receptor interaction may reflect an initial event in the action of PGF₂α as a luteolytic agent.     

The cyanobacterium Synechococcus R-2 (Anacystis nidulans, S. leopoliensis) PCC 7942 has a sodium-dependent chloride transporter

Synechococcus R-2 (PCC 7942) actively accumulated Cl^? in the light and dark, under control conditions (BG-11 media: pH_o, 7·5; [Na⁺]_o, 18 mol m^?3; [Cl^?]_o, 0·508 molm^?3). In BG-11 medium [Cl^?], was 17·2±0·848 mol m^?3 (light), electrochemical potential of Cl^? (ΔμCl^?_i,o) =+211±2mV; [Cl^?]_i= 1·24±0·11 mol m^?3(dark), ΔμCl^?_i,o=+133±4mV. Cl^? fluxes, but not permeabilities, were much higher in the light: ?Cl^?_i,o= 4·01±5·4 nmol m^?2 s^?1, ^PCl^?_i,o= 47±5pm s^?1 (light); ?Cl^?_i,o= 0·395±0·071 nmol m^?2 s^?1, _PCl^?_i,o= 69±14 pm s^?1 (dark). Chloride fluxes are inhibited by acid pH_o (pH_o 5; ?Cl^?_i,o= 0·14±0·04 nmol m^?2 s^?1); optimal at pH_o 7·5 and not strongly inhibited by alkaline pH_o (pH_o 10; ?Cl^?1_i,o= 1·7±0·14 nmol m^?2 s^?1). A Cl^?_in/2H⁺_in coporter could not account for the accumulation of Cl^? alkaline pH_o. Permeability of Cl^? is very low, below 100pm s^?1 under all conditions used, and appears to be maximal at pH_o 7·5 (50–70 pm s^?1) and minimal in acid pH_o (20pm s^?1). DCCD (dicyclohexyl-carbodiimide) inhibited ?Cl^?_i,o in the light about 75% and [Cl^?]_i fell to 2·2±0·26 (4) mol m^?3. Valinomycin had no effect but monensin severely inhibited Cl^? uptake ([Cl^?]_i= 1·02±0·32 mol m^?3; ?Cl^?_i,o= 0·20±0·1 nmol m^?2 s^?1). Vanadate (200 mmol m^?3) accelerated the Cl^? flux (?Cl^?_i,o= 5·28±0·64 nmol m^?2 s^?1) but slightly decreased accumulation of Cl^? ([Cl^?], = 13·9±1·3 mol m^?3) in BG-11 medium but had no significant effect in Na⁺-free media. DCMU (dichlorophenyldimethylurea) did not reduce [Cl^?], or ?Cl^?_i,o to that found in the dark ([Cl^?]_i= 8·41±0·76 mol m^?3; ?Cl^?_i,o= 2·06±0·36 nmol m^?2 s^?1). Synechococcus also actively accumulated Cl^? in Na⁺-free media, [Cl^?]_i was lower but ΔΨ_i,o hyperpolarized in Na⁺-free media and so the ΔμCl^?_i,o was little changed ([Cl^?]_i= 7·98±0·698 mol m^?3; ΔμCl^?_i,o=+203±3 mV). Net Cl^? uptake was stimulated by Na⁺; Li⁺ acted as a partial analogue for Na⁺. Synechococcus has a Na⁺ activated Cl^? transporter which is probably a primary 2Cl^?/ATP pump. The Cl^? pump is voltage sensitive. ΔμCl^?_i,o is directly proportional to ΔΨ_i,o(P»0·01%): ΔμCl^?_i,o= -1·487 (±0·102) ×ΔΨ_i,o, r= -0·983, n= 31. The ΔμCl^?_i,o increased (more positive) as the Δμ_i,o became more negative. The ΔμCl^?_i,o has no known function, but might provide a driving force for the uptake of micronutrients.     

Metal cation toxicity in the alga Gracilaria domingensis as evaluated by the daily growth rates in synthetic seawater

Macroalgae of the genus Gracilaria have considerable economic importance as raw material for agar production and belong to an important group of organisms that are tolerant of high concentrations of metal. The median inhibitory concentration (IC₅₀) values obtained by measuring the ratio of fresh mass variation (i.e., daily growth rates) of the red macroalga Gracilaria domingensis during a 48-h aquatic toxicity assay are reported here. The alga was exposed to 14 different metal cations as well as the molybdate anion in synthetic seawater. The actual concentrations of these ionic species (at IC₅₀ values) and the proportion of free ions (aqueous complexes) were determined by inductively coupled plasma atomic emission spectroscopy and the Environmental Protection Agency-recommended software, MINTEQA2, respectively. Based on the free IC₅₀ values (IC₅₀ ^F), the ions were ranked in terms of toxicity: Cd²⁺???Cu²⁺???Pb²⁺???Zn²⁺???Ni²⁺?>?Co²⁺?>?La³⁺???Mn²⁺?>?Ca²⁺?~?Li⁺???MoO₄ ^2????Sr²⁺?>?Mg²⁺???K⁺?>?Na⁺. As a member of the first trophic level in the marine food chain, G. domingensis is an appropriate target organism both for the development of toxicological assays and as a bioindicator of marine degradation.     

Nitrite and Nitrate Levels in Individual Molluscan Neurons: Single-Cell Capillary Electrophoresis Analysis

Abstract: Cell and tissue concentrations of NO₂^? and NO₃^? are important indicators of nitric oxide synthase activity and crucial in the regulation of many metabolic functions, as well as in nonenzymatic nitric oxide release. We adapted the capillary electrophoresis technique to quantify NO₂^? and NO₃^? levels in single identified buccal neurons and ganglia in the opisthobranch mollusc Pleurobranchaea californica, a model system for the study of the chemistry of neuron function. Neurons were injected into a 75-µm separation capillary and the NO₂^? and NO₃^? were separated electrophoretically from other anions and detected by direct ultraviolet absorbance. The limits of detection for NO₂^? and NO₃^? were <200 fmol (<4 µM in the neurons under study). The NO₂^? and NO₃^? levels in individual neurons varied from 2 mM (NO₂^?) and 12 mM (NO₃^?) in neurons histochemically positive for NADPH-diaphorase activity down to undetectable levels in many NADPH-diaphorase-negative cells. These results affirm the correspondence of histochemical NADPH-diaphorase activity and nitric oxide synthase in molluscan neurons. NO₂^? was not detected in whole ganglion homogenates or in hemolymph, whereas hemolymph NO₃^? averaged 1.8 ± 0.2 × 10^?3M. Hemolymph NO₃^? in Pleurobranchaea was appreciably higher than values measured for the freshwater pulmonate Lymnaea stagnalis (3.2 ± 0.2 × 10^?5M) and for another opisthobranch, Aplysia californica (3.6 ± 0.7 × 10^?4M). Capillary electrophoresis methods provide utility and convenience for monitoring NO₂^?/NO₃^? levels in single cells and small amounts of tissue.     

Metal complexes of cyclic diamines,dissociation kinetics of bis(1,5-diazacyclo-octane)nickel(II) in acidic and basic solution,and electrochemical studies

The preparation of the planar yellow [Ni([8]aneN₂)₂](ClO₄)₂ is described. The complex dissociates in basic solution, with rate = k_OH[NiL][OH^?] (L = 1,5-diazacyclo-octane). At 25 °C, k_OH = 4.5 x 10^?2 M^?1 s^?1 and the corresponding activation parameters are ΔH^≠ = 69.2 kJ mol^?1 and ΔS₂₉₈^≠ = ?38.6 J K^?1 mol^?1. Acid catalysed dissociation in quite slow even in strongly acidic solutions. The kinetic data in this case can be fitted to the expression K_obs = k_o + K_H[H⁺], where k_o relates to a solvolytic pathway and k_H to the acid catalysed pathway. At 60 °C, K_o = 2 x 10^?5 s^?1 and k_H is 2 x 10^?5 M^?1 s^?1. Possible mechanisms for these reactions are considered.The Ni(II)/Ni(III) redox couple for NiLⁿ⁺ is irreversible on Pt using MeCN as solvent.