Experimental installation for measurements of chlorophyll fluorescence,CO2 exchange,and transpiration of a detached leaf

An original experimental installation is described to illustrate the unit-based principle in the open-mode design construction of a modern system for the combined fluorometric and CO₂/H₂O gas exchange measurements in the studies of the light and dark reactions of photosynthesis and the transpiration of a detached leaf.     

Simultaneous measurement of nitrogen fixation estimated by acetylene-ethylene assay and nitrate absorption by soybeans

An apparatus was designed for simultaneous measurement of rates of N₂ fixation estimated by C₂H₂-C₂H₄ assay (N₂[C₂H₂] fixation) and NO₃⁻ absorption by roots of intact, nodulated soybeans (Glycine max [L.] Merr.). The principal design features include: (a) a gas-tight mist chamber in which nodulated roots can be exposed simultaneously to C₂H₂ in the gas phase and to a liquid phase containing NO₃⁻ sprayed in a fine mist; and (b) provision for sampling the gas phase for C₂H₄ determination, and the liquid phase for NO₃⁻ determination.     

Clean technology in aquaculture — a production without waste products?

A method has been developed for the determination of H₂S and FeS in sediments. FeS is converted into H₂S which is flushed from the samples directly into an excess of chlorine bleach, NaC1O or KClO with some Zn²⁺ added. Either the excess can be titrated back potentiometrically with As₂O₃, or the sulphate formed can be measured colorimetrically. The precision is primarily controlled by the homogeneity of the sediment suspensions and can be better than 99%.     

Characterizing and modeling hydrogen sulfide production in anaerobic digestion of livestock manure,agro-industrial wastes,and wastewater sludge

Hydrogen sulfide (H₂S) is the most undesirable inorganic gas in biogas from anaerobic digestion (AD). However, H₂S production in AD is complex and understanding of its processes is still limited. This study performed six controlled batch anaerobic co-digestion experiments to investigate H₂S production. Materials were obtained from four field anaerobic digester systems and co-digestion feedstocks from agroindustry. An additional precipitation experiment was conducted to further examine H₂S production dynamics. Digesters containing highly soluble, carbohydrate-based wastes had a high H₂S final specific production (FSP) value. Additionally, the FSP values were negatively correlated with the initial Fe(II):S ratios in the digester liquid of the batch tests. The precipitation experiment indicated that iron sulfide precipitation was preferred in the presence of an anaerobic community. The H₂S production as a time series was successfully modeled using a generalized additive model (R² > 0.82). This study revealed that sulfate, phosphorus, and iron concentrations are important predictors and potential inhibitors of H₂S production in AD. Further examination of real-time H₂S modeling in AD is warranted.     

The Relationship between H(2) Evolution and Acetylene Reduction in Pisum sativum-Rhizobium leguminosarum Symbioses Differing in Uptake Hydrogenase Activity

Peas (Pisum sativum L.) were inoculated with strains of Rhizobium leguminosarum having different levels of uptake hydrogenase (Hup) activity and were grown in sterile Leonard jars under controlled conditions. Rates of H₂ evolution and acetylene reduction were determined for intact nodulated roots at intervals after the onset of darkness or after removal of the shoots. Hup activity was estimated using treatment plants or equivalent plants from the growth chamber, by measuring the uptake of H₂ or ³H₂ in the presence of acetylene. In all cases, the rate of H₂ evolution was a continuous function of the rate of acetylene reduction. In symbioses with no demonstrable Hup activity, H₂ evolution increased in direct proportion to acetylene reduction and the slopes were similar with the Hup⁻ strains NA502 and 128C79. Hup activity was similar in strains 128C30 and 128C52 but significantly lower in strain 128C54. With these strains, the slopes of the H₂ evolution versus acetylene reduction curves initially increased with acetylene reduction, but became constant and similar to those for the Hup⁻ strains at high rates of acetylene reduction. On these parallel portions of the curves, the decreases in H₂ evolution by Hup⁺ strains were similar in magnitude to their H₂-saturated rates of Hup activity. The curvilinear relationship between H₂ evolution and acetylene reduction for a representative Hup⁺ strain (128C52) was the same, regardless of the experimental conditions used to vary the nitrogenase activity.     

Synthesis and reactivity of binuclear halo-bridge palladium(II) isocyanide complexes

An equimolecular mixture of [Pd(RNC)₂Cl₂] (R = Ph, p-Me C₆H₄) and [Pd(MeCN)₂Cl₂] reacts in boiling, 1,2-dichloroethane to give the binuclear complexes [Pd(RNC)Cl₂]₂.These compounds undergo a variety of bridge-splitting reactions with neutral or anionic ligands yielding complexes of the type cis and trans [Pd(RNC)LX₂] or [Pd(RNC)X₃]⁻ (L = PPh₃, pyridine, C₆H₁₁NC; X = CL, Br).By reaction of [Pd(PhNC)Cl₃]⁻ with MeOH the anionic carbene complex [Pd{C(NHPh)OMe}Cl₃]⁻ is obtained.[Pd(PhNC)Cl₂]₂ reacts with p-toluidine (excess) or o-aminopyridine to give the corresponding mononuclear carbene derivatives.In the case of the mixed derivative [Pd(p-MeC₆H₄NC)(C₆H₁₁NC)Cl₂], only the more activated p-tolylisocyanide was found to react with p-toluidine.The complexes have been characterized by elemental analysis, conductivity measurements, i.r. and ¹H n.m.r. spectra where possible.     

Changes in ion fluxes and the energy demand during spore development inPhytophthora infestans zoospores

Cell-free supernatant of pelleted zoospores was found to be more suitable for maintaining viable zoospores and developed cysts than the supernatant of mature cysts. Conductivity and pH measurements indicated quantitative changes in the ionic composition of a suspension ofP. infestans zoospores during their conversion into cysts. An increase in conductivity in the incipient cyst suspension was followed by a decrease of conductivity in the maturing cyst suspension. The conductivity changes correlated closely with K⁺ fluxes which, in turn, coincided with the reverse, but stoichiometrically smaller, H⁺ fluxes. Zoospores treated with 1.5 μmol/L DCCD (an inhibitor of plasma membrane H⁺-ATPase) or 100 mmol/L Li⁺ (an inhibitor of cell motility) released predominantly K⁺ and other cations and their O₂ consumption decreased. The H⁺/K⁺ exchange is therefore very probably associated with an operation of the plasma membrane H⁺-ATPase. The differential decrease in respiration caused by DCCD and Li⁺ was used to estimate the energy demand for cell motility and spore development.     

Thermodynamic and kinetic analysis of the H2 threshold for Methanobacterium bryantii M.o.H

H₂ thresholds, concentrations below which H₂ consumption by a microbial group stops, have been associated with microbial respiratory processes such as dechlorination, denitrification, sulfate reduction, and methanogenesis. Researchers have proposed that observed H₂ thresholds occur when the available Gibbs free energy is minimal (ΔG ≈ 0) for a specific respiratory reaction. Others suggest that microbial kinetics also may play a role in controlling the thresholds. Here, we comprehensively evaluate H₂ thresholds in light of microbial thermodynamic and kinetic principles. We show that a thermodynamic H₂ threshold for Methanobacterium bryantii M.o.H. is not controlled by ΔG for methane production from H₂ + HCO₃⁻. We repeatedly attain a H₂ threshold near 0.4 nM, with a range of 0.2–1 nM, and ΔG for methanogenesis from H₂ + HCO₃⁻ is positive, +5 to +7 kJ/mol-H_2, at the threshold in most cases. We postulate that the H₂ threshold is controlled by a separate reaction other than methane production. The electrons from H₂ oxidation are transferred to an electron sink that is a solid-phase component of the cells. We also show that a kinetic threshold (S _min) occurs at a theoretically computed H₂ concentration of about 2400 nM at which biomass growth shifts from positive to negative.     

Enzyme immunosensor. III. Amperometric determination of human chorionic gonadotropin by membrane-bound antibody.

An enzyme immunosensor was constructed for the determination of human chorionic gonadotropin (HCG), which is a hormone and an important diagnostic measure of pregnancy. An antibody to HCG was immobilized to a membrane. The antibody-bound membrane was placed onto an oxygen probe so as to react with HCG either specifically or selectively. Catalase, which catalyzes the decomposition of H₂O₂ into H₂O and O₂, was used to label HCG. Nonlabeled HCG to be assayed and catalase-labeled HCG were competitively reacted with the membrane-bound antibody of the sensor to form an antigen-antibody complex on the membrane surface. After the removal of nonspecifically adsorbed HCG, the sensor was contacted with a H₂O₂ solution. The membrane-adsorbed catalase enzymatically generated oxygen with a resulting increase in cathodic current of the sensor. The HCG concentration was determined from the initial rate of the current increase. The enzyme immunosensor was applied to the determination of HCG in the concentration range of 2 × 10^?2 to 10² IU/ml.     

Gas Metabolism Evidence in Support of the Juxtaposition of Hydrogen-Producing and Methanogenic Bacteria in Sewage Sludge and Lake Sediments

We developed new techniques to measure dissolved H₂ and H₂ consumption kinetics in anoxic ecosystems that were not dependent on headspace measurements or gas transfer-limited experimentation. These H₂ metabolism parameters were then compared with measured methane production rates, and estimates of H₂ production and interspecies H₂ transfer were made. The H₂ pool sizes were 205 and 31 nM in sewage sludge from an anaerobic digestor and in sediments (24 m) from Lake Mendota, respectively. The H₂ turnover rate constants, as determined by using in situ pool sizes and temperatures, were 103 and 31 h⁻¹ for sludge and sediment, respectively. The observed H₂ turnover rate accounted for only 5 to 6% of the expected H₂-CO₂-dependent methanogenesis in these ecosystems. Our results are in general agreement with the results reported previously and are used to support the conclusion that most of the H₂-dependent methanogenesis in these ecosystems occurs as a consequence of direct interspecies H₂ transfer between juxtapositioned microbial associations within flocs or consortia.     

Inferred relatedness and heritability in malaria parasites

Malaria parasites vary in phenotypic traits of biomedical or biological interest such as growth rate, virulence, sex ratio and drug resistance, and there is considerable interest in identifying the genes that underlie this variation. An important first step is to determine trait heritability (H²). We evaluate two approaches to measuring H² in natural parasite populations using relatedness inferred from genetic marker data. We collected single-clone Plasmodium falciparum infections from 185 patients from the Thailand–Burma border, monitored parasite clearance following treatment with artemisinin combination therapy (ACT), measured resistance to six antimalarial drugs and genotyped parasites using 335 microsatellites. We found strong relatedness structure. There were 27 groups of two to eight clonally identical (CI) parasites, and 74 per cent of parasites showed significant relatedness to one or more other parasites. Initially, we used matrices of allele sharing and variance components (VC) methods to estimate H². Inhibitory concentrations (IC₅₀) for six drugs showed significant H² (0.24 to 0.79, p = 0.06 to 2.85 × 10⁻⁹), demonstrating that this study design has adequate power. However, a phenotype of current interest—parasite clearance following ACT—showed no detectable heritability (H² = 0–0.09, ns) in this population. The existence of CI parasites allows the use of a simple ANOVA approach for quantifying H², analogous to that used in human twin studies. This gave similar results to the VC method and requires considerably less genotyping information. We conclude (i) that H² can be effectively measured in malaria parasite populations using minimal genotype data, allowing rational design of genome-wide association studies; and (ii) while drug response (IC₅₀) shows significant H², parasite clearance following ACT was not heritable in the population studied.     

Measurement of the extent of electron transfer to the bacteriopheophytin in the M-subunit in reaction centers of Rhodopseudomonas viridis

We have measured the extent of flash-induced electron transfer from the bacteriochlorophyll dimer, P, to the bacteriopheophytin in the M-subunit, H_M, in reaction centers of Rhodopseudomonas viridis. This has been done by measuring the transient states produced by excitation of reaction centers trapped in the PH_L ^–H_M state at 90 K. Under these conditions the normal forward electron transfer to the bacteriopheophytin in the L-subunit, H_L, is blocked and the yield of transient P⁺H_M ^– can be estimated with respect to the lifetime of P^*. Under these conditions flash induced absorbance decreases of the bacteriochlorophyll dimer 990 nm band suggest that a transient P⁺ state is formed with a quantum yield of 0.09±0.06 compared to that formed during normal photochemistry. These transient measurements provide an upper limited on the yield of a transient P⁺ H_M ^– state. An estimate of 0.09 as the yield of the P⁺ H_M ^– state is consistent with all current observations. This estimate and the lifetime of P^* suggest that the electron transfer rate from P^* to H_M, k_M, is about 5 × 10⁹ sec^–1 (_M = 200ps). These measurements suggest that the a branching ratio k_L/k_M is on the order of 200. The large value of the branching ratio is remarkable in view of the structural symmetry of the reaction center. This measurement should be useful for electron transfer calculations based upon the reaction center structure.     

Effect of pH on H-2H exchange,H2 production and H2 uptake,catalysed by the membrane-bound hydrogenase of Paracoccus denitrificans

(1) The kinetics of isotope exchange catalysed by the membrane-bound hydrogenase of Paracoccus denitrificans have been studied by measuring H²H, H₂ or ²H₂ produced when the enzyme catalyses the exchange between ²H₂ and H₂O or H₂ and ²H₂O. (2) In the ²H₂-H₂O system the measured rate of H₂ production was always higher than that of H²H. The $\text{H}{}_2\text{H}{}^2\text{H}$ ratio remained constant (about 1.70) in the protein concentration range 0.08–1.32 mg. The very rapid formation of H₂ with respect to H²H is consistent with the hypothesis of a heterolytic cleavage of ²H₂ into a deuteron and an enzyme hydride that can exchange with the solvent. (3) In the H₂-²H₂O system, the exchange rate was much lower than in the ²H₂-H₂O system, indicating a marked isotopic effect of ²H₂O. (4) The H-²H exchange activity, determined from the initial velocity of H²H formation, is optimal at pH 4.5. A second maximum of activity is observed at pH 8.3. The pH value of 4.5 is also the pH optimum for H₂ production while at pH 8.3–8.5 there is a maximum of H₂ oxidation activity. (5) In ordinary H₂O the K_m for hydrogen uptake estimated either from H₂ consumption or from benzyl viologen reduction was 0.06–0.07 μM for both H₂ and ²H₂ indicating a strong affinity of the enzyme for hydrogen at pH 8.3–8.5. Shifting from H₂O to ²H₂O does not affect the K_m of the enzyme for H₂ but lowers the V_max value about 10-fold. The K_m for benzyl viologen and methyl viologen was 0.08 and 2 mM, respectively.     

Physiological effects of long term exposure to low concentrations of SO2 and NH3 on poplar leaves

Shoots of poplar (Populus euramericana L. cv. Flevo) were exposed to filtered air, SO₂, NH₃ or a mixture of SO₂ and NH₃ for 7 weeks in fumigation chambers. After this exposure gas exchange measurements were carried out using a leaf chamber. As compared to leaves exposed to filtered air, leaves pretreated with 112 μg m^?3 SO₂ showed a small reduction in maximum CO₂ assimilation rate (P_max) and stomatal conductance (g_s). They also showed a slightly higher quantum yield and dark respiration. In addition, the fluorescence measurements indicated that the Calvin cycle of the leaves pretreated with 112 μg m^?3 SO₂ was more rapidly activated after transition from dark to light. An exposure to 64 μg m^?3 NH₃ had a positive effect on P_max, stomatal conductance and NH₃ uptake of the leaves. This positive effect was counteracted by an SO₂ concentration of 45 μg m^?3. The exposure treatments appeared to have no effect on the relationship between net CO₂-assimilation and g_s. Also, no injury of the leaf cuticle or of epidermal cells was observed. Resistance analysis showed that NH₃ transfer into the leaf can be estimated from data on the boundary layer and stomatal resistance for H₂O transfer and NH₃ concentration at the leaf surface, irrespective of whether the leaves are exposed for a short or long time to NH₃ or to a mixture of NH₃ and SO₂. In contrast SO₂ uptake into the leaves was only partly correlated to the stomatal resistance. The results suggest a large additional uptake of this gas by the leaves. The possibility of a difference in path length between SO₂ and H₂O molecules is proposed.     

Effect of Dy3+ or Eu2+ co‐activator on a BaCa(SO4)2:Ce3+ mixed alkaline earth sulfate phosphor

The individual emission and energy transfer between Ce³⁺ and Eu²⁺ or Dy³⁺ in BaCa(SO₄)₂ mixed alkaline earth sulfate phosphor prepared using a co‐precipitation method is described. The phosphor was characterized by X‐ray diffraction (XRD) and photoluminescence (PL) studies and doped by Ce;Eu and Dy rare earths. All phosphors showed excellent blue–orange emission on excitation with UV light. PL measurements reveal that the emission intensity of Eu²⁺ or Dy³⁺ dopants is greater than when they are co‐doped with Ce³⁺. An efficient Ce³⁺ → Eu²⁺ [²T_2g(4f⁶5d) → ⁸S_7/2(4f₇)] and Ce³⁺ → Dy³⁺ (⁴ F_9/2 → ⁶H_15/2 and ⁴ F_9/2 → ⁶H_13/2) energy transfer takes place in the BaCa(SO₄)₂ host. A strong blue emission peak was observed at 462 nm for Eu²⁺ ions and an orange emission peak at 574 nm for Dy³⁺ ions. Hence, this phosphor may be used as a lamp phosphor. Copyright © 2015 John Wiley & Sons, Ltd.     

Inducing salt tolerance in maize (Zea mays L.) through seed priming with chloride salts: Growth and ion transport at early growth stages

The study was conducted to determine whether salt tolerance could be induced in maize at germination stage by soaking of seeds for 8 h in distilled water or in 200 meq·L⁻¹ of NaCl, KCl, CaCl₂·2H₂O. Both primed and un-primed seeds were subjected for 14 days to 0, 100 or 200 mol·m⁻³ NaCl under controlled conditions. Although all priming agents were effective in alleviating adverse effects of salt stress on maize at germination stage, CaCl₂·2H₂O proved to be more effective since the seeds primed with this salt had significantly higher final germination, rate of germination and fresh and dry weights of plumules and radicles than those treated with other salts or distilled water. Concentration of Na⁺, K⁺ and Ca²⁺ increased significantly in all parts of germinating seeds of maize seeds primed with NaCl, KCl, or CaCl₂·2H₂O, respectively. In addition, seeds primed with CaCl₂·2H₂O were the highest in Cl⁻ accumulation in all parts of the germinating seeds, followed by seeds treated with NaCl and KCl. Most of the Ca²⁺ was retained in seeds and mesocotyl, because of which, transport of this ion to plumules and radicles was low.     

The quantum requirement for H2 production by anoxygenic phototrophic bacteria

Cultures from three groups of phototrophic bacteria (green sulphur bacteria, purple non-sulphur bacteria and purple sulphur bacteria) were investigated in respect of the quantum requirement for H₂ production (Q_H2). Rates of H₂ formation were determined by means of a Clark-type H₂ electrode with dense suspensions of whole cells and malate, acetate or sulphide as electron donor. At low light intensities (0–3 W·m^–2 of monochromatic light) the minimum quantum requirement was 8.6 quanta per H₂ with Chlorobium vibrioforme, 7.5 with Rhodospirillum rubrum, and 23.2 with Ectothiorhodospira shaposhnikovii. The physiological efficiency, defined as the measured Q_H2 compared to the theoretical value calculated from the energy requirement of the physiological processes involved, was 94%, 88%, or 28%, respectively. With increasing light intensities the quantum requirement also increased. Various hydrogenase inhibitors either inhibited both H₂ uptake and production (Cu²⁺, NO), or affected neither of these activities (CO, C₂H₂, N₂O, ethylenedinitrilotetraacetate). An uptake hydrogenase-deficient Hup^–-mutant of R. rubrum had higher rates of net H₂ production but a similar quantum requirement. The energetic efficiency of H₂ production by various biological and artificial systems is discussed.     

Identification and characterization of plasmids in hydrogen uptake positive and hydrogen uptake negative strains ofRhizobium japonicum

Modifications were made of published procedures to allow routine isolation of plasmids fromRhizobium japonicum. The plasmid profiles of a series of H₂ uptake positive and H₂ uptake negative strains were compared. None of the strains ofR. japonicum with high H₂ uptake activities exhibited discernible plasmids, while most of the strains, with little or no H₂ uptake activity, showed plasmids with molecular weights ranging from approximately 49–290 x10⁶. An examination of H₂ uptake negative mutants derived from an H₂ uptake positive parent revealed two discernible plasmid bands in nonrevertible mutants but no detectable plasmids in revertible mutants or in the parent strain from which mutants were derived.     

18O composition of CO2 and H2O ecosystem pools and fluxes in a tallgrass prairie: Simulations and comparisons to measurements

In this paper we describe measurements and modeling of ¹⁸O in CO₂ and H₂O pools and fluxes at a tallgrass prairie site in Oklahoma. We present measurements of the δ¹⁸O value of leaf water, depth‐resolved soil water, atmospheric water vapor, and Keeling plot δ¹⁸O intercepts for net soil‐surface CO₂ and ecosystem CO₂ and H₂O fluxes during three periods of the 2000 growing season. Daytime discrimination against C¹⁸OO, as calculated from measured above‐canopy CO₂ and δ¹⁸O gradients, is also presented. To interpret the isotope measurements, we applied an integrated land‐surface and isotope model (ISOLSM) that simulates ecosystem H₂¹⁸O and C¹⁸OO stocks and fluxes. ISOLSM accurately predicted the measured isotopic composition of ecosystem water pools and the δ¹⁸O value of net ecosystem CO₂ and H₂O fluxes. Simulations indicate that incomplete equilibration between CO₂ and H₂O within C₄ plant leaves can have a substantial impact on ecosystem discrimination. Diurnal variations in the δ¹⁸O value of above‐canopy vapor had a small impact on the predicted δ¹⁸O value of ecosystem water pools, although sustained differences had a large impact. Diurnal variations in the δ¹⁸O value of above‐canopy CO₂ substantially affected the predicted ecosystem discrimination. Leaves dominate the ecosystem ¹⁸O‐isoflux in CO₂ during the growing season, while the soil contribution is relatively small and less variable. However, interpreting daytime measurements of ecosystem C¹⁸OO fluxes requires accurate predictions of both soil and leaf ¹⁸O‐isofluxes.     

Novel Sub‐5 nm Layered Niobium Phosphate Nanosheets for High‐Voltage,Cation‐Intercalation Typed Electrochemical Energy Storage in Wearable Pseudocapacitors

Layered niobium phosphates have been considered very promising energy storage materials because of their high theoretical operating voltage window and the rich oxidation states of niobium. However, their development has been stymied by the phase‐controlled synthesis due to the insolubility of niobium sources except in concentrated hydrofluoric (HF) acid systems. Herein, a new avenue is opened for layered acid niobium phosphate (2NbOPO₄·H₃PO₄·H₂O) synthesis in a mild oxalic acid system. Taking advantage of this strategy, in situ growth of sub‐5 nm 2NbOPO₄·H₃PO₄·H₂O nanosheet (NPene) arrays on conductive carbon fiber cloth (CFC) substrates is achieved as self‐standing electrodes for solid‐state supercapacitors. Interestingly, the NPene@CFC electrode exhibits a typical cation (H⁺ or Li⁺)‐intercalation kinetics with a wide potential window of 0–1.0 V in aqueous electrolytes. Given the wide potential window and highly exposed active surface, the solid‐state asymmetric supercapacitors constructed from such a NPenes@CFC electrode display a high working potential of 2.0 V, energy density of 122.2 W h kg^?1 at a power density of 589.7 W kg^?1, cycle stability with a capacitance retention of 94.2% after 10 000 cycles, and also outstanding flexible and wearable characteristics.