The effect of chronic ethanol consumption on temperature-dependent physical properties of liver mitochondrial membranes

We have reported that the monovalent ionophore monensin causes undersulfated chondroitin sulfate biosynthesis in cultured chondrocytes. In order to clarify the mechanism of this diminished sulfation, we have measured the rate of incorporation of sulfate into chondrocytes and assayed the cellular ATP levels. We have also measured sulfatase activity, the incorporation of ³⁵SO₄ into 3′-phosphoadenosine 5′-phospho[³⁵S]sulfate and endogenous sulfotransferase activity in the cell-free extracts. We find that: (1) The incorporation of ³⁵SO₄ into the free sulfate pool in chondrocytes was not inhibited by monensin. (2) The ATP levels of monensin-treated chondrocytes were the same as control cells. (3) There was no sulfatase activity in both control and monensin-treated chondrocytes. (4) Enzymatic analyses revealed that ³⁵SO₄ incorporation into 3′-phosphoadenosine 5′-phospho[³⁵S]sulfate and subsequent sulfotransferase activity were not inhibited in the presence of monensin. At present the most tenable hypothesis to account for monensin causing undersulfated chondroitin sulfate synthesis is that the ionophore impairs the access of proteoglycans to the sulfotransferases in the luminal walls of the Golgi structures.     

Localization of sulfate reduction in planted and unplanted rice field soil

Rates of in situ sulfate reduction (SRR) in planted and unplanted rice fieldsoil were measured by the ³⁵SO^2– ₄-radiotracermethod using soil microcosms. The concentration of ³⁵SO^2– ₄ decreased exponentially with time.However, time course experiments indicated that incubation times of10–30 min were appropriate for measurements of SRRusing a single time point in routine assays. Unplanted microcosmsshowed high SRR of 177 nmol cm^-3 d^-1 inthe uppermost centimeter where average sulfate concentrations were<33 µM. Fine scaled measurements (1 mmresolution) localized highest SRR (<100 nmol cm^-3d^-1) at the oxic/anoxic interface at 2–5 mmdepth. In planted rice field soil, SRR of <310 nmolcm^-3 d^-1 were observed at 0–2cm depth. Sulfate reduction rates were determined at a millimeter-scalewith distance to a two dimensional root compartment. The SRR was highestat 0–1.5 mm distance to the root layer with rates up to500 nmol cm^-3 d^-1, indicating a highstimulation potential of the rice roots. SRR seemed to be mainlydependent on the in situ sulfate porewater concentrations. At thesoil surface of unplanted microcosms sulfate concentration decreasedfrom <150 µM to <10 µM within the first 8 mm of depth. In planted microcosmssulfate concentration varied from 87–99 µMsulfate at the 0–3 mm distance to the root layer to48–62 µM sulfate at a root distance>4 mm from the roots.The depth distribution of inorganic sulfur compounds was determinedfor planted and unplanted rice field soil. Sulfate, acid volatilesulfide (AVS) and chromium reducible sulfide (CRS) were up to 20 foldhigher in planted than in unplanted microcosms. CRS was the majorinsoluble sulfur fraction with concentrations >1.7µmol cm^-3. Organic sulfur accounted for25–46% of the total sulfurpresent (269 µg/g dw) in an unplanted microcosm.The biogeochemical role of sulfate reduction forshort-term accumulation of inorganic sulfur compounds(FeS, FeS_{_2} and S°) in rice soil wasdetermined in a time course experiment with incubationperiods of 5, 10, 20, 30 and 60 min. The relativedistribution of CRS and AVS formation showedlittle depth dependence, whereas the formation of³⁵S° seemed to be the highest in themore oxidized upper soil layers and near the root surface.AV³⁵S was the first major product of sulfatereduction after 20–30 min, whereas CR³⁵Swas formed, as AV³⁵S and ³⁵S°decreased, at longer incubation periods of >30 min.     

Cycling of inorganic and organic sulfur in peat from Big Run Bog,West Virginia

Total S concentration in the top 35 cm of Big Run Bog peat averaged 9.7 mol·g — wet mass^–1 (123 mol·g dry mass^–1). Of that total, an average of 80.8% was carbon bonded S, 10.4% was ester sulfate S, 4.5% was FeS₂­S, 2.7% was FeS­S, 1.2% was elemental S, and 0.4% was SO₄ ^2–­S. In peat collected in March 1986, injected with³⁵S­SO₄ ^2– and incubated at 4 °C, mean rates of dissimilatory sulfate reduction (formation of H₂S + S⁰ + FeS + FeS₂), carbon bonded S formation, and ester sulfate S formation averaged 3.22, 0.53, and 0.36 nmol·g wet mass^–1·h^–1, respectively. Measured rates of sulfide oxidation were comparable to rates of sulfate reduction. Although dissolved SO₄ ^2– concentrations in Big Run Bog interstitial water (< 200 µM) are low enough to theoretically limit sulfate reducing bacteria, rates of sulfate reduction integrated throughout the top 30–35 cm of peat of 9 and 34 mmol·m^–2·d^–1 (at 4 °C are greater than or comparable to rates in coastal marine sediments. We suggest that sulfate reduction was supported by a rapid turnover of the dissolved SO₄ ^2– pool (average turnover time of 1.1 days). Although over 90% of the total S in Big Run Bog peat was organic S, cycling of S was dominated by fluxes through the inorganic S pools.     

Sulfate reduction and S-oxidation in a moorland pool sediment

In an oligotrophic moorland pool in The Netherlands, S cycling near the sediment/water boundary was investigated by measuring (1) SO₄ ^2– reduction rates in the sediment, (2) depletion of SO₄ ^2– in the overlying water column and (3) release of³⁵S from the sediment into the water column. Two locations differing in sediment type (highly organic and sandy) were compared, with respect to reduction rates and depletion of SO₄ ^2– in the overlying water.Sulfate reduction rates in sediments of an oligotrophic moorland pool were estimated by diagenetic modelling and whole core³⁵SO₄ ^2– injection. Rates of SO₄ ^2– consumption in the overlying water were estimated by changes in SO₄ ^2– concentration over time in in situ enclosures. Reduction rates ranged from 0.27–11.2 mmol m^–2 d^–1. Rates of SO₄ ^2– uptake from the enclosed water column varied from –0.5, –0.3 mmol m^–2 d^–1 (November) to 0.43–1.81 mmol m^–2 d^–1 (July, August and April). Maximum rates of oxidation to SO₄ ^2– in July 1990 estimated by combination of SO₄ ^2– reduction rates and rates of in situ SO₄ ^2– uptake in the enclosed water column were 10.3 and 10.5 mmol m^–2 d^–1 at an organic rich and at a sandy site respectively.Experiments with³⁵S^2– and³⁵SO₄ ^2– tracer suggested (1) a rapid formation of organically bound S from dissimilatory reduced SO₄ ^2– and (2) the presence of mainly non SO₄ ^2–-S derived from reduced S transported from the sediment into the overlying water. A³⁵S^2– tracer experiment showed that about 7% of³⁵S^2– injected at 1 cm depth in a sediment core was recovered in the overlying water column.Sulfate reduction rates in sediments with higher volumetric mass fraction of organic matter did not significantly differ from those in sediments with a lower mass fraction of organic matter.Corresponding author     

Assay of R-apomorphine, S-apomorphine, apocodeine, isoapocodeine and their glucuronide and sulfate conjugates in plasma and urine of patients with Parkinson's disease

Analytical methods are described for the selective, rapid and sensitive determination of R- and S-apomorphine, apocodeine and isoapocodeine and the glucuronic acid and sulfate conjugates in plasma and urine. The methods involve liquid-liquid extraction followed by high-performance liquid chromatography with electrochemical detection. The glucuronide and sulfate conjugates are determined after enzymatic hydrolysis. For the assay of R- and S-apomorphine a 10 μm Chiralcel OD-R column is used and the voltage of the detector is set at 0.7 V. The mobile phase is a mixture of aqueous phase (pH 4.0)-acetonitrile (65:35, v/v). At a flow-rate of 0.9 ml min⁻¹ the total run time is ca. 15 min. The detection limits are 0.3 and 0.6 ng ml⁻¹ for R- and S- apomorphine, respectively (signal-to-noise ratio 3). The intra- and inter-assay variations are <5% in the concentration range of 2.5-25 ng ml⁻¹ for plasma samples, and <4% in the concentration range of 40-400 ng ml⁻¹ for urine samples. For the assay of apomorphine, apocodeine and isoapocodeine, a 5 μm C₁₈ column was used and the voltage of the detector set at 0.825 V. Ion-pairing chromatography was used. The mobile phase is a mixture of aqueous phase (pH 3.0)-acetonitrile (75:25, v/v). At a flow-rate of 0.8 ml min⁻¹ the total run time is ca. 14 min. The detection limits of this assay are 1.0 ng ml⁻¹ for apomorphine and 2.5 ng ml⁻¹ for both apocodeine and isoapocodeine (signal-to-noise ratio 3). The inter-assay variations are 5% in the concentration range of 5-40 ng ml⁻¹ for plasma samples and 7% in the concentration range of 50-500 ng ml⁻¹ for urine samples. The glucuronic acid and sulfate conjugates of the various compounds are hydrolysed by incubation of the samples with β-glucuronidase and sulfatase type H-1, respectively. Hydrolysis was complete after 5 h of incubation. No measurable degradation of apomorphine, apocodeine and isoapocodeine occurred during the incubation. A pharmacokinetic study of apomorphine, following the intravenous infusion of 30 μg kg⁻¹ for 15 min in a patient with Parkinson's disease, demonstrates the utility of the methods: both the pharmacokinetic parameters of the parent drug and the appearance of apomorphine plus metabolites in urine could be determined.     

Anti-inflammatory drugs, prostanoid and proteoglycan production by cultured bovine articular chondrocytes

The effect of various anti-inflammatory drugs on the production of prostaglandins E₂ and F₂α, 6 keto PGF₁α and thromboxane B₂ by bovine articular chondrocytes was measured by radioimmunoassay. While indomethacin and meclofenamic acid caused a dose-dependent inhibition of all prostanoids measured, the effects of hydrocortisone and colchicine varied with respect to different prostanoids. Hydrocortisone (10⁻⁷M – 10⁻³M) both in the presence and absence of added arachidonic acid, resulted in an inhibition of prostaglandins E₂ and F₂, and to a lesser extent, 6 keto PGF₁α, but T×B₂ production was only slightly inhibited by the drug in the absenced of arachidonic acid and markedly increased in its presence. Colchicine (10⁻⁷M – 10⁻³M) had the opposite effect, causing an inhibition of T×B₂ and stimulating PGE₂ and 6 keto PGF₁α production. These findings suggest that certain anti-inflammatory drugs may, in addition to their action on phospholipase A₂ and cyclo-oxygenase, exert potent effects at the level of the different synthetases. In order to see whether these alterations in relative prostanoid levels affected proteoglycan metabolism, the effect of anti-inflammatory drugs on proteoglycan synthesis by cultured chondrocytes was tested using ³⁵SO₄ labeling methodology. The results showed that the concentrations tested (10⁻⁵M to 10⁻⁷M), indomethacin, dexamethasone, hydrocortisone and colchicine inhibited ³⁵SO₄ incorporation into newly synthesized proteoglycan molecules both in the presence (10⁻⁶M) and absence of exogenous arachidonic acid. In the same concentration range choroquine had no effect.These results do not support the hypothesis of direct prostanoid involvement in the modulation of proteoglycan synthesis in articular cartilage.     

Temperature dependence of anion transport in the human red blood cell

Arrhenius plots of chloride and bromide transport yield two regions with different activation energies (E_a). Below 15 or 25°C (for Cl⁻ and Br⁻, respectively), E_a is about 32.5 kcal/mol; above these temperatures, about 22.5 kcal/mol (Brahm, J. (1977) J. Gen. Physiol. 70, 283–306). For the temperature dependence of SO₄²⁻ transport up to 37°C, no such break could be observed. We were able to show that the temperature coefficient for the rate of SO₄²⁻ transport is higher than that for the rate of denaturation of the band 3 protein (as measured by NMR) or the destruction of the permeability barrier in the red cell membrane. It was possible, therefore, to extend the range of flux measurements up to 60°C and to show that, even for the slowly permeating SO₄²⁻ in the Arrhenius plot, there appears a break, which is located somewhere between 30 and 37°C and where E_a changes from 32.5 to 24.1 kcal/mol. At the break, the turnover number is approx. 6.9 ions/band 3 per s. Using ³⁵Cl⁻-NMR (Falke, Pace and Chan (1984) J. Biol. Chem. 259, 6472–6480), we also determined the temperature dependence of Cl⁻-binding. We found no significant change over the entire range from 0 to 57°C, regardless of whether the measurements were performed in the absence or presence of competing SO₄²⁻. We conclude that the enthalpy changes associated with Cl⁻-or SO₄²⁻-binding are negligible as compared to the E_a values observed. It was possible, therefore, to calculate the thermodynamic parameters defined by transition-state theory for the transition of the anion-loaded transport protein to the activated state for Cl⁻, Br⁻ and SO₄²⁻ below and above the temperatures at which the breaks in the Arrhenius plots are seen. We found in both regions a high positive activation entropy, resulting in a low free enthalpy of activation. Thus the internal energy required for carrying the complex between anion and transport protein over the rate-limiting energy barrier is largely compensated for by an increase of randomness in the protein and/or its aqueous environment.     

Dynamics of Bacterial Sulfate Reduction in a Eutrophic Lake

Bacterial sulfate reduction in the surface sediment and the water column of Lake Mendota, Madison, Wis., was studied by using radioactive sulfate (³⁵SO₄²⁻). High rates of sulfate reduction were observed at the sediment surface, where the sulfate pool (0.2 mM SO₄²⁻) had a turnover time of 10 to 24 h. Daily sulfate reduction rates in Lake Mendota sediment varied from 50 to 600 nmol of SO₄²⁻ cm⁻³, depending on temperature and sampling date. Rates of sulfate reduction in the water column were 10³ times lower than that for the surface sediment and, on an areal basis, accounted for less than 18% of the total sulfate reduction in the hypolimnion during summer stratification. Rates of bacterial sulfate reduction in the sediment were not sulfate limited at sulfate concentrations greater than 0.1 mM in short-term experiments. Although sulfate reduction seemed to be sulfate limited below 0.1 mM, Michaelis-Menten kinetics were not observed. The optimum temperature (36 to 37°C) for sulfate reduction in the sediment was considerably higher than in situ temperatures (1 to 13°C). The response of sulfate reduction to the addition of various electron donors metabolized by sulfate-reducing bacteria in pure culture was investigated. The degree of stimulation was in this order: H₂ > n-butanol > n-propanol > ethanol > glucose. Acetate and lactate caused no stimulation.     

Sulfur cycling in the water column of Little Rock Lake, Wisconsin

The S cycle in the water column of a small, soft-water lake was studied for 9 years as part of an experimental study of the effects of acid rain on lakes. The two basins of the lake were artificially separated, and one basin was experimentally acidified with sulfuric acid while the other served as a reference or control. Spatial and seasonal patterns of sulfate uptake by plankton (53–70 mmol m^–2 yr^–1), deposition of sulfur to sediments in settling seston (53 mmol m^–2 yr^–1), and sulfate diffusion (0–39 mmol m^–2 yr^–1) into sediments were examined. Measurements of inputs (12–108 mmol m^–2 yr^–1) and outputs (5.5–25 mmol m^–2 yr^–1) allowed construction of a mass balance that was then compared with rates of S accumulation in sediments cores (10–28 mmol m^–2 yr^–1) and measured fluxes of S into the sediments. Because of the low SO₄ ^2– concentrations (µmole L^–1) in the lake, annual uptake by plankton (53–70 mmol m^–2 yr^–1) represented a large fraction (>50%) of the SO₄ ^2– inventory in the lake. Despite this large flux through the plankton, only small seasonal fluctuations in SO₄ ^2– concentrations (µmole L^–1) were observed; rapid mineralization of organic matter (half-life <3 months) prevented sulfate depletion in the water column. The turnover time for sulfate in the water column is only 1.4 yr; much less than the 11-yr turnover time of a conservative ion in this seepage lake. Sulfate diffusion into and reduction in the sediments (0–160 µmole m^–2 d^–1) caused SO₄ ^2– depletion in the hypolimnion. Modeling of seasonal changes in lake-water SO₄ ^2– concentrations indicated that only 30–50% of the diffusive flux of sulfate to the sediments was permanently incorporated in solid phases, and about 15% of sulfur in settling seston was buried in the sediments. The utility of sulfur mass balances for seepage lakes would be enhanced if uncertainty about the deposition velocity for both sulfate aerosols and SO₂, uncertainty in calculation of a lake-wide rate of S accumulation in sediments, and uncertainty in the measured diffusive fluxes could be further constrained.     

Reduction of Sulfur Compounds in the Sediments of a Eutrophic Lake Basin

Concentrations of various sulfur compounds (SO₄²⁻, H₂S, S⁰, acid-volatile sulfide, and total sulfur) were determined in the profundal sediments and overlying water column of a shallow eutrophic lake. Low concentrations of sulfate relative to those of acid-volatile sulfide and total sulfur and a decrease in total sulfur with sediment depth implied that the contribution of dissimilatory sulfur reduction to H₂S production was relatively minor. Addition of 1.0 mM Na₂³⁵SO₄ to upper sediments in laboratory experiments resulted in the production of H₂³⁵S with no apparent lag. Kinetic experiments with ³⁵S demonstrated an apparent K_m of 0.068 mmol of SO₄²⁻ reduced per liter of sediment per day, whereas tracer experiments with ³⁵S indicated an average turnover time of the sediment sulfate pool of 1.5 h. Total sulfate reduction in a sediment depth profile to 15 cm was 15.3 mmol of sulfate reduced per m² per day, which corresponds to a mineralization of 30% of the particulate organic matter entering the sediment. Reduction of ³⁵S⁰ occurred at a slower rate. These results demonstrated that high rates of sulfate reduction occur in these sediments despite low concentrations of oxidized inorganic compounds and that this reduction can be important in the anaerobic mineralization of organic carbon.     

Regulation of sulfate uptake and xylem loading of poplar roots (Populus tremula x P. alba)

Sulfate transport processes and its regulation were studied in roots of poplar trees (Populus tremula x P. alba). From the exponential increase in sulfate uptake with temperature an activation energy (E_a) of 9.0±0.8 kJ mol^–1 was calculated. In the concentration range 0.005–10 mM sulfate uptake showed biphasic Michaelis-Menten kinetics with a K_m of 3.2±3.4 M and a V_max of 49±11 nmol SO₄^2– g^–1 FW h^–1 for the high-affinity uptake system (phase 1) and a K_m of 1.33±0.41 mM and a V_max of 255±25 nmol SO₄^2– g^–1 FW h^–1 for the low-affinity system (phase 2). Xylem loading decreased linearly with temperature and remained unchanged within the sulfate concentration range studied. Regulation of sulfate uptake and xylem loading by O-acetyl serine (OAS), Cys, reduced glutathione (GSH), Met and S-methylmethionine (SMM) were tested by perfusion into the xylem sap with the pressure probe and by addition to the incubation medium. When added directly to the transport medium, Cys and GSH repressed, and OAS stimulated sulfate uptake; xylem loading was stimulated by Cys, repressed by GSH and only slightly affected by OAS. When perfused into the xylem, none of the compounds tested affected sulfate uptake of excised roots, but xylem loading was stimulated by SMM and OAS and repressed by Met. Apparently, the site of application strongly determined the effect of regulatory compounds of sulfate transport processes.     

Effect of ions on phospholipid layer structure as indicated by Raman Spectroscopy

Various anions and cations are found to induce changes in the layered structure of phosphatidylcholine-water systems as indicated by Raman Spectroscopy. From the ratio of Raman intensities, , it is inferred that dispositive ions decrease the proportion of gauche character in the hydrocarbon chains, with the relative influence being: Ba²⁺ < Mg²⁺ < Ca²⁺ ≈ Cd²⁺. Unipositive ions (Li⁺, K⁺ and Na⁺) produce no observed changes in the Raman spectrum of the lecithin dispersion. The proportion of gauche character of the hydrocarbon chains is found to be nearly independent of the anion for: Br⁻, Cl⁻, acetate⁻, I⁻, ClO₄⁻, CNS⁻ and SO₄²⁻. Dispersions prepared with a solution of KI + I₂ produced Raman spectra in which the 1089 cm⁻¹ peak, which is characteristic of random lipid chains, was greatly intensified, presumably because of the presence of I₃⁻ which is known to penetrate the lipid lamellae. The observed trends are discussed.     

Acid hydrolysis of sugarcane bagasse for lactic acid production

In order to use sugarcane bagasse as a substrate for lactic acid production, optimum conditions for acid hydrolysis of the bagasse were investigated. After lignin extraction, the conditions were varied in terms of hydrochloric (HCl) or sulfuric (H₂SO₄) concentration (0.5–5%, v/v), reaction time (1–5 h) and incubation temperature (90–120 °C). The maximum catalytic efficiency (E) was 10.85 under the conditions of 0.5% of HCl at 100 °C for 5 h, which the main components (in g l⁻¹) in the hydrolysate were glucose, 1.50; xylose, 22.59; arabinose, 1.29; acetic acid, 0.15 and furfural, 1.19. To increase yield of lactic acid production from the hydrolysate by Lactococcus lactis IO-1, the hydrolysate was detoxified through amberlite and supplemented with 7 g l⁻¹ of xylose and 7 g l⁻¹ of yeast extract. The main products (in g l⁻¹) of the fermentation were lactic acid, 10.85; acetic acid, 7.87; formic acid, 6.04 and ethanol, 5.24.     

Non-aqueous capillary electrophoresis chiral separations with sulfated β-cyclodextrin

This paper reports the application of an anionic cyclodextrin (CD), sulfated β-cyclodextrin with a degree of substitution of four (β-CD-(SO₄⁻)₄, in chiral separations of pharmaceutical enantiomers by non-aqueous capillary electrophoresis (NACE). Upon complexation with the anionic CD, electrophoretic mobilities of the basic enantiomers decreased, however, both separation selectivity and resolution were enhanced. The advantage of NACE chiral separations over the aqueous CE with the charged CD is that higher electric field strength and higher ionic strength could be applied due to the characteristics of the solvent formamide. The higher ionic strength leads to stacking of peaks and reduces the electrodispersion caused by the mobility mismatch between β-CD-(SO₄⁻)₄–analyte complexes and the co-ions in the running buffer. As a result, better peak shapes and higher separation efficiency were obtained. Comparing with NACE chiral separations with neutral CDs, lower concentration of β-CD-(SO₄⁻)₄ was needed due to the fact that the electrostatic attraction caused stronger binding between β-CD-(SO₄⁻)₄ and the enantiomers. The effects of the experimental parameters, such as concentration of the CD, apparent pH (pH*), degree of substitutions of the CDs, percentage of water in mixed solvent systems, and type of solvents were also studied.     

Disruption of sulfur cycling and acid neutralization in lakes at low pH

Experimental acidification of a softwater lake to below pH 5 fundamentally changed the sulfur cycle and lowered internal alkalinity generation (IAG). Prior to reaching pH 4.5, the balance of sulfur reduction and oxidation reactions within the lake was in favour of reduction, and the lake was a net sink for sulfate. In the four years at pH 4.5 the balance of reduction and oxidation reactions was in favour of oxidation, and there was a net production of sulfate (SO₄ ^2–) within the lake. Evidence indicating a decrease in net SO₄ ^2– reduction at pH 4.5 was also obtained in an anthropogenically acidified lake that had been acidified for many decades. In both lakes, the decrease in net SO₄ ^2– reduction appeared to be linked not to a simple inhibition of SO₄ ^2– reduction but rather to changes in benthic ecosystem structure, especially the development of metaphytic filamentous green algae, which altered the balance between SO₄ ^2– reduction and sulfur oxidation.At pH's above 4.5, net SO₄ ^2– reduction was the major contributor to IAG in the experimental lake, as it is in many previously studied lakes at pH 5 and above. At pH 4.5, the change in net annual SO₄ ^2– reduction (a decrease of 110%) resulted in a 38% decrease in total IAG. Because of the important role of net SO₄ ^2– reduction in acid neutralization in softwater lakes, models for predicting acidification and recovery of lakes may need to be modified for lakes acidified to pH <5.     

Partitioning of mercury in aqueous biphasic systems and on ABEC™ resins

Poly(ethylene glycol)-based aqueous biphasic systems (PEG-ABS) can be utilized to separate and recover metal ions in environmental and hydrometallurgical applications. A concurrent study was conducted comparing the partitioning of mercury between aqueous layers in an ABS [Me-PEG-5000/(NH₄)₂SO₄] and partitioning of mercury from aqueous solutions to aqueous biphasic extraction chromatographic (ABEC-5000) resins. In ammonium sulfate solutions, mercury partitions to the salt-rich phase in ABS, but by using halide ion extractants, mercury will partition to the PEG-rich phase after formation of a chloro, bromo or iodo complex. The efficacy of the extractant increases in the order Cl⁻<Br⁻<I⁻. This behavior is also observed using the ABEC resins where halo complexes of mercury will adsorb to the resin from (NH₄)₂SO₄ solutions with retention following the same order. The onset of mercury extraction or adsorption is different for the three extractants, occurring at the lowest extractant concentration for I⁻, followed by Br⁻, and then Cl⁻. Fluoride does not extract mercury. Extraction or adsorption of mercury is improved at the lowest halide concentrations in the presence of sulfuric acid. The addition of sulfuric acid to (NH₄)₂SO₄ solution results in ABEC retention of mercury even in the absence of halide extractant.     

Effect of rice plants on methane production and rhizospheric metabolism in paddy soil

In order to elucidate the effects of rice plants on CH₄ production, we conducted experiments with soil slurries and planted rice microcosms. Methane production in anoxic paddy soil slurries was stimulated by the addition of rice straw, of unsterile or autoclaved rice roots, and of the culture fluid in which rice plants had axenically been cultivated. The addition of these compounds also increased the concentrations of acetate and H₂, precursors of CH₄ production, in the soil. Planted compared to unplanted paddy soil microcosms exhibited lower porewater CH₄ concentrations but higher CH₄ emission rates. They also exhibited higher sulfate concentrations but similar nitrate concentrations. Concentrations of acetate, lactate and H₂ were not much different between planted and unplanted microcosms. Pulse labeling of rice plants with¹⁴CO₂ resulted during the next 5 days in transient accumulation of radioactive lactate, propionate and acetate, and after the second day of incubation in the emission of¹⁴CH₄. Most of the radioactivity (40–70%) was incorporated into the above-ground biomass of rice plants. However, during a total incubation of 16 days about 3–6% of the applied radioactivity was emitted as¹⁴CH₄, demonstrating that plant-derived carbon was metabolized and significantly contributed to CH₄ production. The sequence of the appearance of radioactive products and their specific radioactivities indicate that CH₄ was produced from root exudates by a microbial community consisting of fermenting and methanogenic bacteria.     

Kinetic Studies of Bacterial Sulfate Reduction in Freshwater Sediments by High-Pressure Liquid Chromatography and Microdistillation

Indirect photometric chromatography and microdistillation enabled a simultaneous measurement of sulfate depletion and sulfide production in the top 3 cm of freshwater sediments to be made. The simultaneous measurement of sulfate depletion and sulfide production rates provided added insight into microbial sulfur metabolism. The lower sulfate reduction rates, as derived from the production of acid-volatile ³⁵S²⁻ only, were explained by a conversion of this pool to an undistillable fraction under acidic conditions during incubation. A mathematical model was applied to calculate sulfate reduction from sulfate gradients at the sediment-water interface. To avoid disturbance of these gradients, the sample volume was reduced to 0.2 g (wet weight) of sediment. Sulfate diffusion coefficients in the model were determined (D_s = 0.3 × 10⁻⁵ cm² s⁻¹ at 6°C). The results of the model were compared with those of radioactive sulfate turnover experiments by assessing the actual turnover rate constants (2 to 5 day⁻¹) and pool sizes of sulfate at different sediment depths.     

Versicolorin A hemiacetal,hydroxydihydrosterigmatocystin, and aflatoxin G2a reductase activity in extracts fromAspergillus parasiticus

Versicolorin A hemiacetal was converted to versicolorin C in cell-free systems fromAspergillus parasiticus. The rate of reaction catalyzed by the 35–70% ammonium sulfate fraction was 0.43 nmol min^–1 mg^–1 with NADPH as cosubstrate and 0.17 nmol. min^–1 mg^–1 with NADH at 25°C at pH 7.4. The product from incubation of 17-hdyroxy-16,17-dihydrosterigmatocystin with the 35–70% ammonium sulfate fraction and NADPH was a polar compound which was converted to dihydrosterigmatocystin by 0.4 M HCl. The olar comound is proposed to be the 14,17-hydrated open-chain derivative of dihydrosterigmatocystin. Aflatoxin G_2a was also reduced in this system to a polar product tentatively identified as the 13,16-hydrated open-chain derivative of AFG₂. The reductase activity may be involved in the formation of reduced intermediates and aflatoxins in cultures ofA. parasiticus.     

Sulfate reduction in fine-grained sediments in the Eastern Scheldt,southwest Netherlands

Sulfate reduction and sulfide accumulation were examined in fine-grained sediments from rapidly accreting abandoned channels and mussel culture areas in the Eastern Scheldt, which covered 4 and 5% of the total surface area, respectively.Reduction rates were measured in batch experiments in which the SO₄ ^2– depletion was measured during anoxic incubation. The reduction rates in summer varied between 14–68 mmol SO₄ ^2– m^–2 day^–1 and were related to the sedimentation rate. In the most rapidly accreting channels, SO₄ ^2– was exhausted below 15–50 cm and methanogenesis became the terminal process of organic carbon oxidationOne-dimensional modelling of sulfate profiles in mussel banks indicated that the subsurface influx of SO₄ ^2– was almost of the same order as the diffusive flux at the sediment-seawater interface, during the initial stages of the mussel bank accretion. The energy dissipation of waves and tidal currents on the mussel bank surface increased the apparent sediment diffusivity up to 3-fold, especially in the winterThe results indicate that acid volatile sulfide (AVS) was the major, in-situ reduced, sulfur compound in the sediment. The sulfidation of easily extractable iron was nearly complete. Pyrite concentrations (40–80 M S cm^–3) were as high as the AVS concentrations, but there was apparently no in-situ transformation of AVS into pyrite. The detrital pyrite originated from eroding marine sediments elsewhere