Catalytic properties of three lactate dehydrogenases from potato tubers (Solanum tuberosum)

Two l-lactate dehydrogenase isoenzymes and one dl-lactate dehydrogenase could be separated from potato tubers by polyacrylamide-gel electrophoresis. The enzymes are specific for lactate, while β-hydroxybutyric acid, glycolic acid, and glyoxylic acid are not oxidized. Their pH optima are pH 6.9 for the oxidation and 8.0 for the reduction reaction.The K_m values for l-lactate for the two isoenzymes are 2.00 × 10^?2 and 1.82 × 10^?2, m. In the reverse reaction the affinities for pyruvate are 3.24 × 10^?4 and 3.34 × 10^?4, m. Both enzymes have similar affinities for NAD and NADH (3.00 × 10^?4; 4.00 × 10^?4, and 8.35 × 10^?4; 5.25 × 10^?4, m).The dl-lactate oxidoreductase may transfer electrons either to NAD or N-methyl-phenazinemethosulfate. The K_m values of this enzyme for l-lactate are 4.5 × 10^?2, m and for d-lactate 3.34 × 10^?2, m. Its affinity for pyruvate is 4.75 × 10^?4, m. The enzyme is inhibited by excess NAD (K_m = 1.54 × 10^?4, M) and has an affinity toward NADH (K_m = 5.00 × 10^?3, M) which is about one tenth of that of the two isoenzymes of l-lactate dehydrogenase.     

Feeding efficiency of Chaoborus flavicans (Insecta, Diptera) under turbulent conditions

Turbulence can affect predator–prey interactions. The effect of turbulence on the feeding efficiency of an ambush predator was tested with laboratory experiments. The experiments were conducted in 100-L aquaria in which ten individuals of fourth instar Chaoborus flavicans larvae were placed as predators. Two prey densities (3 and 10 ind. of Daphnia pulex L^?1) and two durations (30 and 120 min) were tested in a nonturbulent treatment and five different turbulence levels [average root-mean-square (RMS) velocities ranging from 0 to 7.3 cm s^?1, corresponding dissipation rates from 7.2 × 10^?7 to 1.3 × 10^?3 m² s^?3]. We hypothesized that the feeding rate of C. flavicans would be enhanced by turbulence due to increasing encounter rates up to a turbulence level above which a disturbance in post-encounter processes would lead to reduced feeding efficiency. However, the results showed no significant increase in the feeding rate of C. flavicans at intermediate turbulence. At high turbulence we found the expected significant negative response in the feeding rate of Chaoborus larvae. The feeding rate declined below the rates at nonturbulent and intermediate turbulence conditions as the average RMS velocity exceeded 3.1 cm s^?1 (dissipation rate 9.9 × 10^?5 m² s^?3, respectively).     

Waste water treatment and metal (Pb<Superscript>2+</Superscript>, Zn<Superscript>2+</Superscript>) removal by microalgal based stabilization pond system

A case study was undertaken for the treatment of domestic wastewater generated at village of Sanghol, Distt. Fatehgarh Sahib, Punjab (India), using a schematic designed algal and duckweed based stabilization pond system, which is discussed here for winter months only (November to March) as there was no growth of duckweeds and only algae dominated the whole system. A proficient increase in pH and dissolved oxygen was observed after the treatment with reduction in chemical oxygen demand and biochemical oxygen demand by 93% and 79% respectively. Chlorella sp. was the dominating algal species in the stabilization pond water during entire period and was studied for its Zn²⁺ and Pb²⁺ metal removal efficiency. 60–70% removal of Zn²⁺ was observed from culture medium containing 5–20 mg L^?1 Zn²⁺, which declined to 42% at 50 mg L^?1. A constant decline in cell number from 538 × 10⁵ to 8 × 10⁵ cells ml^?1 was observed indicating zinc toxicity to Chlorella. Lead was maximally removed by 66.3% from culture medium containing 1 mg L^?1. The lead removal efficiency was 45 50 % at higher 5 to 20 mg L^?1 of external lead concentrations. The increase in cell number indicated no signs of Pb²⁺ toxicity up to 20 mg L^?1. The maximum uptake (q _max) by live Chlorella biomass for both Zn²⁺ and Pb²⁺ was 34.4 and 41.8 mg/g respectively.     

Enhancing Diamond Fluorescence via Optimized Nanorod Dimer Configurations

Light extraction from silicon (SiV) and nitrogen (NV) vacancy diamond color centers coupled to plasmonic silver and gold nanorod dimers was numerically improved. Numerical optimization of the coupled dipolar emitter—plasmonic nanorod dimer configurations was realized to attain the highest possible fluorescence enhancement by simultaneously improving the color centers excitation and emission through antenna resonances. Conditional optimization was performed by setting a criterion regarding the minimum quantum efficiency of the coupled system (cQE) to minimize losses. By comparing restricted symmetric and allowed asymmetric dimers, the advantages of larger degrees of freedom achievable in asymmetric configurations was proven. The highest 2.59?×?10⁸ fluorescence enhancement was achieved with 46.08% cQE via NV color center coupled to an asymmetric silver dimer. This is 3.17-times larger than the 8.19?×?10⁷ enhancement in corresponding symmetric silver dimer configuration, which has larger 68.52% cQE. Among coupled SiV color centers the highest 1.04?×?10⁸ fluorescence enhancement was achieved via asymmetric silver dimer with 37.83% cQE. This is 1.06-times larger than the 9.83?×?10⁷ enhancement in corresponding symmetric silver dimer configuration, which has larger 57.46% cQE. Among gold nanorod coupled configurations the highest fluorescence enhancement of 4.75?×?10⁴ was shown for SiV color center coupled to an asymmetric dimer with 21.8% cQE. The attained enhancement is 8.48- (92.42-) times larger than the 5.6?×?10³ (5.14?×?10²) fluorescence enhancement achievable via symmetric (asymmetric) gold nanorod dimer coupled to SiV (NV) color center, which is accompanied by 16.01% (7.66%) cQE.     

Molecular Detection of Anaerobic Ammonium-Oxidizing (Anammox) Bacteria in High-Temperature Petroleum Reservoirs

Anaerobic ammonium-oxidizing (anammox) process plays an important role in the nitrogen cycle of the worldwide anoxic and mesophilic habitats. Recently, the existence and activity of anammox bacteria have been detected in some thermophilic environments, but their existence in the geothermal subterranean oil reservoirs is still not reported. This study investigated the abundance, distribution and functional diversity of anammox bacteria in nine out of 17 high-temperature oil reservoirs by molecular ecology analysis. High concentration (5.31–39.2 mg l^?1) of ammonium was detected in the production water from these oilfields with temperatures between 55°C and 75°C. Both 16S rRNA and hzo molecular biomarkers indicated the occurrence of anammox bacteria in nine out of 17 samples. Most of 16S rRNA gene phylotypes are closely related to the known anammox bacterial genera Candidatus Brocadia, Candidatus Kuenenia, Candidatus Scalindua, and Candidatus Jettenia, while hzo gene phylotypes are closely related to the genera Candidatus Anammoxoglobus, Candidatus Kuenenia, Candidatus Scalindua, and Candidatus Jettenia. The total bacterial and anammox bacterial densities were 6.4?±?0.5?×?10³ to 2.0?±?0.18?×?10⁶ cells ml^?1 and 6.6?±?0.51?×?10² to 4.9?±?0.36?×?10⁴ cell ml^?1, respectively. The cluster I of 16S rRNA gene sequences showed distant identity (<92%) to the known Candidatus Scalindua species, inferring this cluster of anammox bacteria to be a new species, and a tentative name Candidatus “Scalindua sinooilfield” was proposed. The results extended the existence of anammox bacteria to the high-temperature oil reservoirs.     

Evaluation of the in vitro methods for micropropagation of Chondracanthus acicularis (Roth) Fredericq (Gigartinales, Rhodophyta): tissue culture and production of protoplasts

Tissue was cultured and protoplasts isolated from the carrageenophyte Chondracanthus acicularis with the aim of developing micropropagation as an alternative to harvesting raw material from natural beds. Both adventitious shoots and filamentous calluses were induced by tissue culture on medium solidified with 0.4–1 % (w/v) agar. Adventitious shoots were mainly produced from discoid bases while filamentous calluses were mainly induced from basal zones and sub-apical explants. A gradient of the regeneration ability was observed from the top to the bottom of the thallus. The discoid base was the most reactive explant and produced the highest number of adventitious shoots compared to basal zones and sub-apical explants, irrespective of the concentration of agar. Protoplasts were isolated enzymatically from the whole thallus using a combination of cellulase R-10 Onozuka, macerozyme R-10, and crude extract of the gland gut of algivorous molluscs. The highest mean yield of protoplasts (1.2?×?10⁶ protoplasts g^?1 fresh weight) was obtained after 16 h of digestion with an enzyme mixture containing 2 % (w/v) cellulase R-10, 1 % (w/v) macerozyme R-10 Onozuka, 4 % (v/v) crude extract of gut gland of Haliotis, 0.8 M mannitol, 50 mM sodium citrate, 0.3 % (w/v) bovine serum albumin. Depending on the conditions, mean protoplast yields ranged from 3.14?×?10⁵ to 1.2?×?10⁶ protoplasts g^?1 fresh weight. Different factors (storage duration, mannitol, sodium citrate, crude extract of the gland gut of algivorous molluscs) were tested to improve the yield of protoplasts but none has a significantly effect.     

Lectin-carbohydrate interactions studied by a competitive enzyme inhibition assay

The principle of competitive binding assay in combination with an immobilized lectin (concanavalin A), in close proximity to an oxygen sensor, has been used to quantify carbohydrates and to determine association constants for lectin-carbohydrate interactions. Methyl α-d-mannopyranoside was determined down to 0.5 μg/ml. K_a (maltose) and K_a (maltotriose) was found to be 2.1 × 10³ and 1.7 × 10³m^?1, respectively, which are comparable to values quoted in the literature of approximately 2.8 × 10³m^?1 for both maltose and maltotriose. Furthermore, the estimation of the bonus effect, due to multipoint attachment, for a low-molecular-weight dextran is discussed.     

Combined effect of five single nucleotide polymorphisms related to IL23/Th17 pathway in the risk of psoriasis

Psoriasis is a common immune-mediated inflammatory skin disease with strong genetic components, in which the IL23/Th17 pathway has been implicated. To explore the effective role in psoriasis, we genotyped five single nucleotide polymorphisms in genes related to IL23/Th17 pathway in 14,929 Han Chinese samples. A Bonferroni-corrected significant single-variant association was identified between rs1512970 within IL21 (odds ratio (OR)?=?1.07, 95 % confidence interval (CI)?=?1.02–1.13, P?=?4.94?×?10^?03). We failed to validate rs744166 (OR?=?1.06, 95 % CI?=?1.01–1.11, P?=?1.52?×?10^?02) and other three SNPs (P?=?2.48?×?10^?01?～?1.27?×?10^?02) to meet the single-variant association significance threshold. However, we found that their combined effect substantially contributed to the risk of psoriasis in our sample (P?=?3.91?×?10^?07) and the highest score group conferred the largest risk effect size (OR?=?1.22, 95 % CI?=?1.11–1.34, P?=?1.85?×?10^?05). Our results implicate the ethnic heterogeneity in the susceptibility of psoriasis and suggest common variants with weak effect in IL23/Th17 pathway, which do not show significance in conventional single-variant association study, may contribute to the risk of psoriasis. This study sheds light on the important role of IL23/Th17 pathway in the susceptibility of psoriasis.     

Intramolecular participation of the amide group in the hydrolysis of p-nitrophenyl N-(bromoacetyl) anthranilate

The rate of hydrolysis of p-nitrophenyl N-(bromoacetyl) anthranilate (Ib) has been measured in aqueous solution between pH 1 and 6.5 has been found to increase linearly with pH at pH higher than 3. An abnormally large apparent alkaline rate constant of 3.8 × 10⁶M^?1 sec^?1 has been determined. Intramolecular nucleophilic displacement by the amide group at the carbonyl carbon of the ester occurred and a cyclic intermediate was formed. This intermediate has been detected by direct isolation and by measurements of the proton release accompanying the reaction. The rates of hydrolysis of analogous derivatives (IIb-IIIb-IV), for which this intramolecular assistance was not possible, were slower by a factor of about 5 × 10⁵. Such an example of intramolecular catalysis may be useful for a better understanding of the enzymatic catalysis.     

Response of activated sludge to the treatment of oxytetracycline production waste stream

To investigate how the microbial community in activated sludge responded to high antibiotic levels, a bench-scale aerobic wastewater treatment system was used to treat oxytetracycline (OTC) mother liquor (OTC-ML). Removal efficiency of chemical oxygen demand decreased from 64.9 to 51.0 % when the OTC level increased from 191.6 to 620.5 mg/L, respectively. According to the cloning results, Psychrobacter and Cryptophyta were the dominant bacterium and eukaryote in the inoculated sludge, respectively, both of which related to low temperature. After OTC exposure, Alphaproteobacteria and Betaproteobacteria became the dominant bacteria, with a small proportion of Firmicutes, Actinobacteria appeared, and fungi (mainly Saccharomycotina) became the dominant eukaryotes, indicating the possible functions of these microorganisms in the wastewater treatment of OTC-ML. The relative abundance of nine tetracycline resistance genes and four mobile elements (class 1 integron, class 2 integron, transposon Tn916/1545, and pattern 1 insertion sequence common region) significantly increased from undetectable to 2.1?×?10^?3 in the inoculated sludge to 1.7?×?10^?4–9.8?×?10^?1 in sludge exposed to 620.5 mg/L OTC by using real-time PCR. The variety of gene cassette arrays of class 1 integron in the sludge samples increased with increasing OTC exposure concentration.     

Identification of chromosomal locations associated with tail biting and being a victim of tail-biting behaviour in the domestic pig (Sus scrofa domesticus)

The objective of this study was to identify loci associated with tail biting or being a victim of tail biting in Norwegian crossbred pigs using a genome-wide association study with PLINK case?Ccontrol analysis. DNA was extracted from hair or blood samples collected from 98 trios of crossbred pigs located across Norway. Each trio came from the same pen and consisted of one pig observed to initiate tail biting, one pig which was the victim of tail biting and a control pig which was not involved in either behaviour. DNA was genotyped using the Illumina PorcineSNP60 BeadChip whole-genome single-nucleotide polymorphism (SNP) assay. After quality assurance filtering, 53,952 SNPs remained comprising 74 animals (37 pairs) for the tail biter versus control comparison and 53,419 SNPs remained comprising 80 animals (40 pairs) for the victim of tail biting versus control comparison. An association with being a tail biter was observed on Sus scrofa chromosome 16 (SSC16; p?=?1.6?×?10^?5) and an unassigned chromosome (p?=?3.9?×?10^?5). An association with being the victim of tail biting was observed on Sus scrofa chromosomes 1 (SSC1; p?=?4.7?×?10^?5), 9 (SSC9; p?=?3.9?×?10^?5), 18 (SSC18; p?=?7?×?10^?5 for 9,602,511?bp, p?=?3.4?×?10^?5 for 9,653,881?bp and p?=?5.3?×?10^?5 for 29,577,783 bp) and an unassigned chromosome (p?=?6.1?×?10^?5). An r ²?=?0.96 and a D???=?1 between the two SNPs at 9?Mb on SSC18 indicated extremely high linkage disequilibrium, suggesting that these two markers represent a single locus. These results provide evidence of a moderate genetic association between the propensity to participate in tail-biting behaviour and the likelihood of becoming a victim of this behaviour.     

Development of Beauveria bassiana (Ascomycota: Hypocreales) as a mycoinsecticide to control green peach aphid,Myzus persicae (Homoptera: Aphididae) and investigation of its biocontrol potential

Long-term and intensive use of synthetic insecticides to control the green peach aphid, Myzus persicae (Homoptera: Aphididae) in agricultural production in the world has resulted in pest resistance and environmental pollution. The aim of the study was to develop an environmentally-friendly and effective mycoinsecticide from a local fungal isolate against M. persicae. According to the results of the screening experiments using 15 isolates (6 × Metarhizium, 5 × Beauveria, 2 × Isaria, 2 × Lecanicillium) at 1 × 10⁷ conidia ml^?1 concentrations and the dose–response experiments at 1 × 10⁵ –1 × 10⁹ conidia ml^?1 concentrations against M. persicae nymphs, Beauveria bassiana (KTU-24) was identified as the most promising isolate. The strain KTU-24 is also had tolerance to ecological conditions such as temperature, and UV-B. KTU-24 with advantageous and superior properties was used to develop a test mycoinsecticide. Mass production of spores by KTU-24 was conducted out by liquid-state fermentation using liquid medium. Spores harvested from the sporulated biomass were used to develop an oil-based mycoinsecticide, and the product was designated as AFIDISIDAL-OD Bbas-TR61. The product had lethal effect on M. persicae nymphs at a concentration of 1 × 10⁸ conidia ml^?1 in leaf-disc (82.5%) and pot (84.33%) experiments in a climate chamber. The oil-based mycoinsecticide developed in this study could be profitable when used in aphid-IPM prgrams by reducing crop loss and synthetic pesticides use.     

The hydrolysis of 3-(2-furylacryloyl)-glycyl-l-leucine amide by thermolysin

The kinetics of the hydrolysis of 3-(2-furylacryloyl)-glycycl-l-leucine amide by thermolysin has been reinvestigated. It was found that the K_m for the enzyme substrate interaction is 2.5 × 10^?3m at pH 7.2. This K_m is an order of magnitude less than what has been previously assumed to be the K_m for the enzyme-substrate interaction. The normally recommended assay has 1–3 × 10^?3m substrate and is based on the assumption that the substrate concentration is much less than the K_m. Our data indicate that this assumption appears to be invalid. The hydrolysis of 3-(2-furylacryloyl)-glycyl-l-leucine amide results in a maximum decrease in absorbance at 322 nm. The change in absorbance is nearly 10-fold greater at 322 nm than the change in absorbance at 345 nm where the hydrolysis has been customarily followed. By following the hydrolysis of the substrate at 10^?4m at 322 nm it is possible to work under conditions where the substrate concentration is much less than the K_m.     

Self-exchange rates and electron transfer kinetics of horse heart ferricytochrome C with amino acid-pentacyanoferrate(II) complexes

The electron transfer reactions of horse heart cytochrome c with a series of amino acid-pentacyanoferrate(II) complexes have been studied by the stopped-flow technique, at 25°C, μ = 0.100, pH 7 (phosphate buffer). A second-order behavior was observed in the case of the Fe(CN)₅ (histidine)^3? complex, with k = 2.8 x 10⁵ M^?1 sec^?1. For the Fe(CN)₅ (alanine)^4? and Fe(CN)₅(L-glutamate)^5? complexes, only a minor deviation of the second-order behavior, close to the experimental error (k = 3.2 × 10⁵ and 1.6 x 10⁵ M^?1 sec^?1, respectively) was noted at high concentrations of the reactants (e.g., 6 × 10^?4 M). The results are in accord with recent work on the Fe(CN)₆^4?/cytochrome c system demonstrating weak association of the reactants. The calculated self-exchange rate constants including electrostatic interactions for the imidazole,L -histidine, 4-aminopyridine, glycinate, β-alaninate, andL-glutamate pentacyanoferrate(II) complexes were 3.3 × 105, 3.3 × 10⁵, 2.8 × 10⁶,4.1 × 10²,5.5 × 10², and 6.0 M^?1 sec^?1, respectively. Marcus theory calculations for the cytochrome c reactions were interpreted in terms of two nonequivalent binding sites for the complexes, with the metalloprotein self-exchange rate constants varying from 10⁴ M^?1 sec^?1 (histidine, imidazole, and 4-aminopyridine complexes) to 10⁶ M^?1 sec ^?1 (glycinate, β-alaninate, and L-glutamate complexes).     

Inhibition of ribosomal peptidyltransferase with cytidyl-3' leads to 5'-[2'(3')-O-L-phenylalanyl]-L-adenosine.

The title compound 1e, obtained by chemical synthesis, is an inhibitor of E. coli ribosomal peptidyltransferase. A 50% inhibition of peptidyltransferase-catalyzed N-Ac-Phe-puromycin formation at puromycin concentration 1 × 10^?4 M with 70 S ribosome-poly U-N-Ac[¹⁴C-Phe-tRNA complex occurred at 5 × 10^?4 M of 1e. In contrast, the parent compound 2′(3′)-O-L-phenylalanine-L-adenosine (1b) is a much weaker inhibitor causing only 5% inhibition at 1 × 10^?3 M. Alkaline hydrolysis of compound 1e to cytidylyl-3′→5′-L-adenosine (1c) results in a greatly diminished inhibition which, however, exceeds that of 1b by a factor of two. The inhibition of peptidyltransferase with 1e can be reversed by puromycin. The latter effect levels off at 40% inhibition.     

An energy budget for Simocephalus vetulus (O. F. Muller) (Crustacea : Cladocera)

We discuss the energetics of a cladoceran, Simocephalus vetulus at different temperatures (8.0 ± 1.0, 15.0 ± 1.0, 21.0 ± 1.0 and 28.0 ± 1.0 °C) and food (Chlamydomonas sp.) concentrations (25 × 10³, 50 × 10³, 75 × 10³ and 100 × 10³ cells ml⁻¹). Increase in temperature accelerated ingestion and, to some extent, oxygen consumption. The study revealed a high reproduction efficiency in S. vetulus. Net growth efficiency (ECI) was higher (13.17–41.18%) in pre-adults than in adults (2.71–8.40%). The assimilated energy (A) increased with increasing food concentration at all temperatures. Assimilation efficiency (AD) decreased with increasing food concentrations. The energy used for growth (P) was nearly constant at all food levels because the egested energy increased and assimilation efficiency decreased as food concentration increased.     

Determination of trace amounts of dopamine by flow‐injection analysis coupled with luminol–Ag(III) complex chemiluminescence detection

A novel flow injection analysis‐direct chemiluminescence (FI‐CL) method has been developed for determination of trace amounts of dopamine (DA) based on the enhancing effect of DA on the CL reaction of luminol with an Ag(III) complex in alkaline solution. Under optimum conditions, CL intensities are proportional to the concentration of DA in the range of 1.0 × 10^?10 to 4.0 × 10^?8 mol L^?1. The detection limit is 3.0 × 10^?11 mol L^?1 for DA (3s), with a relative standard deviation (n = 13) of 2.3% for 1.0 × 10^?8 mol L^?1 DA. This method has also been applied for the determination of DA in commercial pharmaceutical injection samples. On the basis of the CL spectra and the results of the free‐radical trapping experiment of this work, a reaction mechanism for this CL reaction is proposed and discussed. Copyright © 2010 John Wiley & Sons, Ltd.     

THE TIME-COURSE OF PHOTOADAPTATION TO LOW-LIGHT IN PROROCENTRUM MARIAE-LEBOURIAE (DINOPHYCEAE)1

The estuarine dinoflagellate, Prorocentrum mariaelebouriae (Parke & Ballantine 1957) Faust 1974 undergoes increases in pigmentation and photosynthetic efficiency within several days of downward light shifts. These changes can be described by first-order kinetics, as has been reported previously for Chlorophyll (Chl) a in several phytoplankton species. The studies described in this paper were conducted with isolates of populations of Prorocentrum from the Chesapeake Bay. We determined rates of adaptation to low-light for cultures grown at a range of photon flux densities (I₀= 2.65–26.2 E.m^?2, d^?1, shifted to 6.3–7.0% I₀) at three temperatures (10°, 15°, and 20° C), bracketing the conditions this species experiences in situ. In this paper, I report the time-course of changes in α, P_max Chl a, peridinin, and I_k and first-order rate constants, K₁ for changes in α, Chl a and peridinin. cell^?1. K₁ for changes in α cell^?1 averaged 1.58 × 10^?2 h^?1 for conditions encompassing five light treatments and three temperatures; the corresponding mean for Chl a was 1.59 × 10^?2 h^?1. Increases in peridinin measured for five light treatments at 15° C showed a mean K₁ of 1.22 × 10^?2 h^?1, Average percent changes in per cell α, Chl a, and peridinin ranged from 0.4–4.0% h^?1 (10–90% d^?1) following exposure to low-light. Photoadaptive changes are important to Prorocentrum because in nature it occupies turbid waters (K_t≥ 0.5 m^?1) where the mixing depth often exceeds the depth of the photic layer. Cells are entrained beneath a seasonally-stable density discontinuity and are exposed to very low-light (< I E.m^?2.d^?1) for days to weeks during subpycnocline transport. The ability of this species to undergo changes in pigmentation and photosynthetic physiology confers increased efficiency of light harvesting and contributes to this species’survival in the estuary where it is an important component of the dinoflagellate flora.     

Kinetic control of co-operativity in the oxygen binding of Panulirus interruptus hemocyanin.

The kinetics of the oxygen reaction of Panulirus interruptus hemocyanin have been studied at pH 9.6 under conditions where the protein exists in the undissociated, co-operative state and in the dissociated, non-co-operative state.Temperature-jump relaxation measurements of the undissociated protein at high oxygen saturation levels show one relaxation process which has been assigned to the high oxygen affinity (R) state, the on and off kinetic constants being 3.1 × 10⁷m^?1s^?1 and 60 s^?1, respectively. Stopped-flow measurements of the oxygen dissociation reaction show (1) an autocatalytic time-course of the reaction at pH 9.6 and (2) an increase in the overall oxygen dissociation rate constant, as the pH is decreased from 9.6 to 7.0.Temperature-jump relaxation measurements of the dissociated protein show one relaxation process characterized by a very high oxygen dissociation rate constant (1500 s^?1) and a combination constant which is of the same order of magnitude as reported for undissociated protein (k_on = 4.6 × 10⁷m^?1s${}^{\mathrm{?1}}$). The behaviour of dissociated protein can be considered as characteristic of the low oxygen affinity (T) state.The results presented in this paper, together with data available for other hemocyanins as well as hemoglobins, lead to the conclusion that respiratory proteins show a common feature in the kinetic control of co-operative oxygen binding: the stability of the oxygen-protein complex is largely determined by the value of the dissociation rate constant, the oxygen combination process very often appearing to be diffusion controlled.     

Vertical distribution of ammonia-oxidizing archaea (AOA) in the hyporheic zone of a eutrophic river in North China

Nitrification plays a significant role in the global nitrogen cycle, and this concept has been challenged with the discovery of ammonia-oxidizing archaea (AOA) in the environment. In this paper, the vertical variations of the diversity and abundance of AOA in the hyporheic zone of the Fuyang River in North China were investigated by molecular techniques, including clone libraries, phylogenetic analysis and real-time polymerase chain reaction. The archaeal amoA gene was detected in all sediments along the profile, and all AOA fell within marine group 1.1a and soil group1.1b of the Thaumarchaeota phylum, with the latter being the dominant type. The diversity of AOA decreased with the sediment depth, and there was a shift in AOA community between top-sediments (0–5 cm) and sub-sediments (5–70 cm). The abundance of the archaeal amoA gene (1.48 × 10⁷ to 5.50 × 10⁷ copies g^?1 dry sediment) was higher than that of the bacterial amoA gene (4.01 × 10⁴ to 1.75 × 10⁵ copies g^?1 dry sediment) in sub-sediments, resulting in a log₁₀ ratio of AOA to ammonia-oxidizing bacteria (AOB) from 2.27 to 2.69, whereas AOB outnumbered AOA in top-sediments with a low log₁₀ ratio of (?0.24). The variations in the AOA community were primarily attributed to the combined effect of the nutrients (ammonium-N, nitrate-N and total organic carbon) and oxygen in sediments. Ammonium-N was the major factor influencing the relative abundance of AOA and AOB, although other factors, such as total organic carbon, were involved. This study helps elucidate the roles of AOA and AOB in the nitrogen cycling of hyporheic zone.