Net Sulfate Research Articles

Seasonal sulfate deposition and export patterns for a small Appalachian watershed

Sulfate deposition and exports from 1988–92 were analyzed for a small headwater catchment in north-central West Virginia. Annual sulfate inputs, estimated by applying throughfall-adjusted ratios to bulk deposition values, and outputs were approximately equal for the five years. Annual mean throughfall-adjusted deposition and export loads were 55.78 and 55.48 kg ha-1, respectively. While these results indicate the watershed has reached sulfate equilibrium relative to current deposition levels, seasonal sulfate accumulations and deficits were evident. Deposition and exports averaged 5.61 and 2.49 kg ha-1 mo-1, respectively, during the growing season, and 3.69 and 5.22 kg ha-1 mo-1 during the dormant season. Sulfur accumulated within the soil during the growing season because inputs of wet and dry sulfur deposition were high while outputs were negligible. The latter was due largely to the lack of runoff resulting from high evapotranspirational demands. By contrast, net sulfate losses occurred during dormant seasons, primarily due to high runoff, even though inputs declined during this season. Researchers working on other watersheds have interpreted similar input/output patterns to mean that sulfate accumulated during the growing season is the source of sulfate exported during the dormant season. However, radioisotopic evidence from a companion study on this watershed showed that some labeled sulfate applied to the watershed more than a year earlier was still present in the organic and mineral soil layers at the point of application (with some as soluble sulfate), and in soil water dispersed throughout the watershed. Its presence indicates that dormant season exports can originate from sulfate deposited over longer periods than just the previous growing season or even previous year. Volume-weighted concentrations in soil leachate suggest that dormant-season sulfate losses resulted from progressive depletion of the anion through the soil profile. During the fall and early winter, soluble sulfate was depleted in the upper soil horizons; in later winter, depletion occurred in the lower horizons.

Calciuric Effects of Short-Term Dietary Loading of Protein, Sodium Chloride and Potassium Citrate in Prepubescent Girls

Objective: Studies using adult human subjects indicate that dietary protein and sodium chloride have negative effects on the retention of calcium by increasing urinary calcium excretion, while alkaline potassium improves calcium retention along with decreasing urinary calcium losses. This study investigated the effect of these dietary factors on acute urinary calcium excretion in 14 prepubescent girls age 6.7 to 10.0 years.Methods: Subjects provided a fasting urine sample then consumed a meal containing one of five treatments: moderate protein (MP) providing 11.8 g protein, moderate protein plus 26 mmol sodium chloride (MP+Na), high protein (HP) providing 28.8 g protein, high protein plus 26 mmol sodium chloride (HP+Na), or high protein plus 32 mmol potassium as tripotassium citrate (HP+K). Urine was collected at 1.5 and 3.0 hours after the meal. Supplemental protein was given as 80:20 casein:lactalbumin. Test meals were isocaloric, and unless intentionally altered, components of interest except phosphate were equal between treatments. Each subject completed all five treatments.Results: Urinary calcium excretion rose after the meal, peaking at 1.5 hours. There were no significant differences in calcium excretion between treatments at any time point. The high protein treatments did not result in a significant increase in either net acid or sulfate excretion at 1.5 hours compared to moderate protein. Dietary sodium chloride had no effect on urinary sodium or calcium excretion over the 3 hours. After the potassium treatment, sodium excretion increased (p≤0.002) and net acid excretion decreased (p<0.001) compared to other treatments at 1.5 hours.Conclusions: In children, a simultaneous increase in protein and phosphorus due to increased milk protein intake did not increase acute urinary calcium excretion. An effect of dietary sodium chloride on acute urinary calcium excretion was not observed. Both these findings were similar to those of adult studies previously conducted in the same laboratory using similar format and treatments. Potassium citrate was not hypocalciuric in children, a response differing from that for adults, who have shown a decrease in acute urinary calcium excretion in response to alkaline potassium treatment. Further characterization of calciuric responses to dietary factors is required for children, who may differ from adults in many respects.

Microsomal steroid sulfatase: interactions with cytosolic steroid sulfotransferases

Net sulfation of 4-methylumbelliferone in intact hepatocytes is regulated, in part, by substrate cycling between sulfotransferases (SULT) and arylsulfatases (ARS). Thus, ARS have the potential to influence rates of net sulfate conjugation of a variety of compounds in intact cells via interaction with SULT. Unlike ARSA and ARSB, which are lysosomal, steroid sulfate sulfatase (ARSC, also known as STS) is localized exclusively in the endoplasmic reticulum (ER). The present study was designed to assess the existence and extent of substrate cycling between steroids and their sulfate conjugates through ARSC and SULT, and also to initiate studies of the topology of the catalytic site of ARSC in the rat liver ER. Addition of rat liver microsomes to cytosol and 3′-phosphoadenosine 5′-phosphosulfate (PAPS) reduced rates of sulfation of dehydroepiandrosterone (DHEA) by SULT, and similarly hydrolysis of DHEA sulfate (DHEAS) was reduced when recombinant human hydroxysteroid SULT was added to rat liver microsomes in the presence of PAPS. There was no evidence for ARSC latency in the presence of detergent at either 4 or 37°C, indicating that facilitated transport of steroid sulfates across the ER membrane may not be required for ARSC activity. The effect of proteases on ARSC activity in intact and disrupted microsomes was determined and compared with effects on components of the glucose-6-phosphatase system known to be localized on the lumenal and cytoplasmic surfaces of the ER. In contrast to the components of the glucose-6-phosphatase system, activity of ARSC in both intact and disrupted microsomes was substantially more resistant to protease inactivation. Our results indicate that substrate cycling of steroids and their sulfates does occur, and suggest that the active site of ARSC may be located within the ER membrane.

Local Sulphide Oxidation Drives Carbonate Dissolution in Modern Shallow Marine Carbonates: Evidence from Oxygen and Sulphur Isotope Composition of Pore Water Sulphate

We have investigated sulphur cycling in modem carbonate sediments from the Florida Platform, U.S.A. Relations among pore water chemistry (SO24 , ZCO2, Ca2+/C1 ) and oxygen and sulphur stable isotope composition of SO 2require a direct coupling between sulphur redox cycling and syndepositional carbonate dissolution. Importantly, despite rapid rates of sulphate reduction, organic-rich carbonate sediment pore waters typically maintain SO2-/C1 ratios within 5% of seawater values, suggesting local reoxidation of H2S. Shallow shelf carbonates comprise 93% of the entire Phanerozoic carbonate rock record and about 38% of the modem oceanic carbonate accumulation, so quantifying sulphur and carbon coupling during the early marine diagenesis is vital in deciphering biogeochemical cycling (e.g. Ronov, 1980; Milliman, 1993). In low-Fe carbonates, H2S-O2 reactions can lower pH values and pore water saturation states promoting carbonate dissolution (e.g. Boudreau, 1991). Given that platform carbonate dissolution fluxes are roughly half of gross biogenic carbonate production, sulphur cycling may greatly affect carbonate preservation and accumulation (Walter and Burton, 1990; Walter et al., 1993). Pore water sulphate ~180 and 834S values were markedly shifted from the established seawater values of +9.5%o (SMOW) and +21.0%o (CDT), respectively. Chemical evolution was least in environments where the sediment-pore water system was relatively 'open' to the overlying seawater. The greatest chemical evolution was observed in bioturbated sediments colonized by Thalassia grass. Here, there was a ~lSOso 4 shift of 5%0, var iab le ~348SO 4 values (+17.7 to +23.3%o), and exceptionally high Ca2+/C1 ratios. These shifts in pore water sulphate isotopic compositions exist despite the apparent lack of net sulphate reduction (Fig. 1). The natural system isotope trend is in marked contrast to the closed system isotope evolution observed in sediment incubation experiments (Fig. 1). Importantly, sulphide oxidation is required as an acid source because pore water Ca+2/C1 and ZCO2 values exceed those attributable to the utilization of dissolved oxygen and the small amount of net sulphate reduction. The observed degree of carbonate dissolution (up to 2.3 mM excess Ca 2+) and sulphate isotopic values are best explained by a redox cycle where bacterial sulphate reduction is followed by efficient H2S oxidation resulting in relatively invariant SO24-/C1 ratios. The 02 required to locally oxidize pore water H2S may come from oxygenated seawater advection or from marine grass rhizome systems which enhance oxygen supply rates. A mass balance model demonstrates that the observed 81SOso4 values are consistent with measured sulphate reduction rates and the short pore water residence times. Pore water geochemistry and sediment incubation experiments confirm that sulphur cycling has a profound effect on the early marine diagenesis of shallow platform carbonates. The important diagenetic processes are oxic respiration, bacterial sulphate reduction, sulphide oxidation, and carbonate dissolution. In most sediments, pore water SO2-/C1 ratios are near overlying ocean values, yet in many cases SO24 has been recycled from HzS. The regularity of pore water evolution and sediment environment becomes more apparent when the oxygen and sulphur isotope compositions of SO 2are examined. Diagenetic effects are most

Posttranscriptional effects of glucose on proteoglycan expression in mesangial cells

Hyperglycemic conditions are known to increase mRNA and protein levels of several extracellular matrix molecules in cultured mesangial cells, but accompanying increases in proteoglycan mRNA have not been found, and there are discrepant reports of normal or decreased proteoglycan synthesis with or without undersulfation in diabetic kidneys and hyperglycemic cultures. We examined the effects in proliferating cells of glucose on [ 35S]sulfate incorporation into heparan and dermatan sulfates and on mRNA levels of decorin, biglycan, and basement membrane perlecan. In both mesangial cells and vascular smooth muscle cells, 30 mmol/L glucose caused a decrease of 15% to 25% in the amount of sulfate incorporated into each proteoglycan in cultures confluent for 1 to 4 days, compared with 10 mmol/L glucose. The effect showed no specificity for the class of proteoglycan and was not a consequence of changes in total protein synthesis, which increased, or cell proliferation, which was unaffected. No decrease in charge density of any of the proteoglycan fractions was observed by ion-exchange chromatography. Therefore, the decrease in labeling was due to a decrease in synthesis and not undersulfation. mRNA levels for biglycan and perlecan increased slightly and transiently, and these changes cannot account for the decreased synthesis. Decorin mRNA was detected only in smooth muscle cells, where it and biglycan were differentially affected by glucose, apparently at the transcriptional level; stabilities of the two messages were unaffected by glucose. Although transforming growth factor-beta 1 (TGF-β 1) mRNA levels increased in response to glucose, the cytokine did not appear to regulate proteoglycan synthesis, because structural changes in proteoglycans elicited by addition of TGF-β 1 to the culture medium did not occur in the hyperglycemic cultures. On the other hand, inhibition and downregulation of protein kinase C (PKC), while decreasing net sulfate incorporation into mesangial cell proteoglycans, prevented the effect of high glucose. We conclude that a high glucose concentration causes a general decrease in the synthesis of all classes of proteoglycans at a posttranscriptional level, and can do so without affecting the charge density of individual proteoglycan molecules.

Hepatic conjugation/deconjugation cycling pathways. Computer simulations examining the effect of Michaelis-Menten parameters, enzyme distribution patterns, and a diffusional barrier on metabolite disposition.

Conjugation/deconjugation cycling plays an important role in the physiologic regulation of the concentration of endogenous compounds that form conjugated metabolites. Less is known concerning the deconjugation of xenobiotics. The model compound p-nitrophenol (pNP) is conjugated to sulfate and glucuronide metabolites which can also undergo hydrolysis, via separate enzyme systems, to regenerate pNP. In the present investigation, computer simulations were performed using literature values for the KM and Vmax for each of the four enzyme systems involved in net pNP conjugation. The apparent sulfation rate, apparent glucuronidation rate, and the extraction ratio (E) of pNP were each examined (i) as a function of pNP concentration, (ii) following alterations in the KM and Vmax values for the deconjugation enzymes, (iii) after modulating the enzyme distribution patterns along the liver flow path for both the conjugating and deconjugating enzymes, and (iv) in the presence of drug metabolite diffusional barriers for membrane transport. Results of these simulations demonstrated that changes in the KM or Vmax for deglucuronidation produced changes not only in net glucuronidation but also in net sulfation. Overall extraction (E) of the parent compound was only affected when glucuronidation was an important pathway, i.e., at higher pNP concentrations. Similar results were observed with changes in desulfation, with desulfation having the greatest effects at low pNP concentrations where sulfation represents the predominant metabolic pathway. Changes in the enzyme distribution patterns for the deconjugation pathways showed that the greatest influence on net conjugation rates occurred when hydrolase enzyme activity was distributed downstream from the respective forward reaction. In the presence of a diffusional barrier for metabolite transport (i.e., when the diffusional clearance was one tenth of blood flow), net metabolism of parent was diminished with E decreasing from 0.74, in the absence of a barrier, to 0.23, since the generated metabolite remained, to a great extent, within hepatocytes and underwent a more pronounced hydrolysis. In the presence of diffusional barriers for uptake of the conjugated metabolites, the lowest drug extraction and metabolite formation rates were observed when the distribution of the conjugation and deconjugation pathways across the liver were the same. Therefore, the effects of deconjugation on hepatic drug removal and metabolite formation are highly dependent on the enzymatic parameters of both the forward and reverse reactions, the parent drug concentration, the enzyme distribution patterns, and the presence of diffusional barriers for metabolite membrane transport. Since a change in the deconjugation of one metabolite can influence the net formation of not only itself but also other metabolites, and overall drug extraction, evaluation of conjugation/deconjugation cycling represents an important consideration in pharmacokinetic studies involving physiological-, pathological-, or pharmacological-induced alterations in conjugate formation.

Effects of sulfate concentration in the overlying water on sulfate reduction and sulfur storage in lake sediments

We investigated the effects of sulfate concentration on sulfate reduction and net S storage in lake sediments using34S as a tracer. The water overlying intact sediment cores from the hypolimnion of Mares Pond, MA, was replaced with two Na234SO4 solutions at either ambient (70 μM) or elevated (260 μM) sulfate concentrations. The δ34S of the added sulfate was 4974 ‰. Over two months, the net sulfate reduction rate in the ambient sulfate treatment was zero, while the net rate for the high sulfate treatment was 140 μmoles/m2/d. The water overlying the cores was kept under oxic conditions and the sediment received no fresh carbon inputs, thus the net rate reported may underestimate the in situ rate. Gross sulfate reduction rates calculated by isotope dilution were approximately 350 μmoles/m2/d for both treatments. While the calculation of gross sulfate reduction rates in intact sediment cores can be complicated by differential diffusion of34S and32S, isotopic fractionation, and the possible formation of ester sulfates, we believe these effects to be small. The results suggest that sulfate reduction is not strongly sulfate-limited in Mares Pond. The difference in net sulfate reduction rates between treatments resulted from a decrease in sulfide oxidation and suggests the importance of reoxidation in controlling net S storage in lake sediments. In both treatments the CRS and organic S fractions were measurably labelled in34S. Below the sediment surface, the CRS fraction was the more heavily labelled storage product for reduced sulfides.

Inorganic sulfur turnover in oligohaline estuarine sediments

Inorganic sulfur turnover was examined in oligohaline (salinity < 2 g kg-1) Chesapeake Bay sediments during the summer. Cores incubated for < 3 hr exhibited higher sulfate reduction (SR) rates (13–58 mmol m-2 d-1) than those incubated for 3–8 hr (3–8 mmol m-2 d-1). SR rates (determined with35SO 4 2- ) increased with depth over the top few cm to a maximum at 5 cm, just beneath the boundary between brown and black sediment. SR rates decreased below 5 cm, probably due to sulfate limitation (sulfate < 25 μM). Kinetic experiments yielded an apparent half-saturating sulfate concentration (Ks) of 34 μM, ≈ 20-fold lower than that determined for sediments from the mesohaline region of the estuary. Sulfate loss from water overlying intact cores, predicted on the basis of measured SR rates, was not observed over a 28-hr incubation period. Reduction of35SO 4 2- during diffusion experiments with intact core segments from 0–4 and 5–9 cm horizons was less than predicted by non-steady state diagenetic models based on35SO 4 2- reduction in whole core injection experiments. The results indicate that net sulfate flux into sediments was an order of magnitude lower than the gross sulfur turnover rate. Solid phase reduced inorganic sulfur concentrations were only 2–3 times less than those in sediments from the mesohaline region of the Bay, despite the fact that oligohaline bottom water sulfate concentrations were 10-fold lower. Our results demonstrate the potential for rapid SR in low salinity estuarine sediments, which are inhabited by sulfate-reducing bacteria with a high affinity for sulfate, and in which sulfide oxidation processes replenish the pore water sulfate pool on a time scale of hours.

Futile cycling between 4-methylumbelliferone and its conjugates in perfused rat liver

Futile cycling between 4-methylumbelliferone and its sulfate and glucuronide conjugates was examined in the single-pass perfused rat liver preparation. The steady-state hepatic extraction ratio of 4-methylumbelliferone was found to be high (0.97) at a low input concentration of 0.005 mumol/L (tracer), with a net 4-methylumbelliferyl sulfate/4-methylumbelliferyl glucuronide ratio of about 5:1; at 63 mumol/L the steady-state extraction ratio had remained constant despite a shift from net sulfation to net glucuronidation. At higher input 4-methylumbelliferone concentrations, saturation was evidenced by a decreased steady-state extraction ratio and reduced net sulfation and net glucuronidation. Because 4-methylumbelliferyl sulfate and 4-methylumbelliferyl glucuronide deconjugation would result in an intracellular accumulation of 4-methylumbelliferone, the phenomenon was monitored with a shift in tracer [3H]4-methylumbelliferone metabolism from sulfation to glucuronidation with increased intracellular 4-methylumbelliferone concentration. When 4-methylumbelliferyl sulfate (0 to 890 mumol/L) or 4-methylumbelliferyl glucuronide (0 to 460 mumol/L) was delivered simultaneously with tracer [3H]4-methylumbelliferone to the rat liver, notable desulfation of 4-methylumbelliferyl sulfate (18% to 38% rate in) but little deglucuronidation of 4-methylumbelliferyl glucuronide (1.2% to 2.1% rate in) was observed. With 4-methylumbelliferyl sulfate, 4-methylumbelliferone and 4-methylumbelliferyl glucuronide were readily found as metabolites, whereas with 4-methylumbelliferyl glucuronide, levels of the metabolites, 4-methylumbelliferone and 4-methylumbelliferyl sulfate, were much reduced. 4-Methylumbelliferyl sulfate and not 4-methylumbelliferyl glucuronide shifted tracer [3H]4-methylumbelliferone metabolism from [3H]4-methylumbelliferyl sulfate to [3H]4-methylumbelliferyl glucuronide formation in a concentration-dependent fashion. The steady-state extraction ratio for 4-methylumbelliferyl sulfate (0.1 to 0.3) was comparatively higher than that for 4-methylumbelliferyl glucuronide (0.05), and it was found to increase with concentration, an observation explained by the nonlinear protein binding of 4-methylumbelliferyl sulfate. Biliary excretion rates for 4-methylumbelliferone and 4-methylumbelliferyl sulfate were proportional to their input or net formation rates, regardless of whether 4-methylumbelliferone, 4-methylumbelliferyl glucuronide or 4-methylumbelliferyl sulfate was administered. By contrast, the excretion rate of 4-methylumbelliferyl glucuronide when administered was only 1/25 the excretion of 4-methylumbelliferyl glucuronide formed from 4-methylumbelliferone and 4-methylumbelliferyl sulfate. The extent of choleresis paralleled the excretion patterns of preformed and formed 4-methylumbelliferyl glucuronide; bile flow was normal with 4-methylumbelliferyl glucuronide administration and was markedly enhanced with increased 4-methylumbelliferone or 4-methylumbelliferyl sulfate administration. The data suggest the presence of a transmembrane barrier for entry of 4-methylumbelliferyl glucuronide and not 4-methylumbelliferyl sulfate or 4-methylumbelliferone into hepatocytes.(ABSTRACT TRUNCATED AT 400 WORDS)

Thermoprotection of a functional epithelium: heat stress effects on transepithelial transport by flounder renal tubule in primary monolayer culture.

Primary monolayer cultures of winter flounder renal proximal-tubule cells were used to determine whether transepithelial transport could be protected from the damaging effects of extreme temperature by previous mild heat shock. Renal tubule epithelial cells were enzymatically dispersed and reorganized as confluent monolayer sheets on native rat tail collagen. Transepithelial electrical properties (potential difference, resistance, short-circuit current, and Na(+)-dependent glucose current) and unidirectional [35S]sulfate fluxes were measured in Ussing chambers at 22 degrees C. Examination of transepithelial electrical properties following acute 1-hr elevation of temperature over a range of 22-37 degrees C provided the basis for the "mild" versus "severe" thermal stress protocols. Severe elevation from 22 degrees C to 32 degrees C for 1.5 hr followed by 1.5 hr at 22 degrees C significantly decreased glucose current (7 +/- 0.7 to 3 +/- 0.8 microA/cm2) as well as net sulfate secretion [131 +/- 11 to 33 +/- 11 nmol/(cm2.hr)]. Mild heat shock of 27 degrees C for 6 hr prior to this severe heat shock completely protected both glucose transport (6 +/- 0.7 microA/cm2) and sulfate flux (149 +/- 13 nmol/(cm2.hr)]. Scanning electron microscopy showed that the number of microvilli on the apical (luminal) surface of the epithelium was decreased after a 32 degrees C heat shock. Monolayers exposed to 27 degrees C for 6 hr prior to incubation at 32 degrees C showed no loss of microvilli. SDS/PAGE analysis of protein patterns from the cultures showed that three classes of heat shock proteins were maximally induced at 27 degrees C. Inhibition of protein synthesis by cycloheximide prevented the thermoprotective effect of mild heat shock. This suggests that certain renal transport functions can be protected from sublethal but debilitating thermal stress by prior mild heat shock and that heat shock proteins may play a role in this protection.

Sulfate transport in human placenta: Further evidence for a sodium-independent mechanism

Sulfate transport in isolated placental brush-border membrane vesicles has properties consistent with an anion exchange process. To ascertain the relevance of this finding to sulfate accumulation by the fetus and placenta in vivo, we examined sulfate transport in human placental tissue slices, comparing sulfate uptake with that of a non-metabolizable amino acid marker, α-aminoisobutyrate (AIB). In contrast to AIB, which was actively concentrated from physiological media, sulfate uptake by the placenta slice was concentrative only in the absence of sodium and at low pH. Uptake of sulfate reached a steady state after 60 min. It was blocked by DIDS (4,4′-diisothiocyanostilbene-2,2′-disulfonate), a specific inhibitor of anion transport, but not by ouabain. We found no evidence for Na +-dependent uptake of sulfate in incubated placental tissue. It seems unlikely that Na +-dependent sulfate transport by the placenta can be responsible for net sulfate accumulation by the human fetus.

Annual cycle of benthic nutrient fluxes in Tomales Bay, California, and contribution of the benthos to total ecosystem metabolism

Benthic fluxes of dissolved nutrients, oxygen, dissolved inorganic carbon, and total alkalinity were measured over a 2 yr period in Tomales Bay, California, USA, using in situ incubation chambers. Release of dissolved nutrients from the sediment peaked in late summer and was lowest in winter. The difference between C:N: P flux ratios and composition of suspended particulates indicated the existence of a sink for regenerated N, relative to C and P. Total alkalinity flux revealed that carbon metabolism by net sulfate reduction represented ca one-third of total benthic metabohsm Partitioning net system fluxes into component fluxes suggested that the equivalent of ca 70 to 80 % of the available particulate C, N and P was respired within the water column, while about 20 to 30 O/O was respired by the benthos. During spring, increasing light resulted in higher water column productivity, followed closely by rising water column respiration. With low delivery of the new organic material to the benthos, and low residual organics in the sediment, benthic respiration remained low. Fallout of particulate material, coinciding with peak water temperature in late summer, resulted in a 'crossover' with benthic respiration temporarily exceeding water column respiration.

Diagenesis in anoxic sediments from the California continental borderland and its influence on iron, sulfur, and magnetite behavior

Solid phase and pore water profiles of compounds containing iron and sulfur have been determined by wet chemical, magnetic, and X‐ray diffraction techniques in three California continental borderland basins. The observed profiles have been fit by simple reaction‐diffusion models in order to determine reaction rates and constrain budgets for iron and sulfur. More than 95% of the solid phase reduced sulfur is pyrite, and down core profiles are well fit by a model in which net sulfate reduction rates decrease exponentially with depth. Net sulfate reduction rates determined from models fit to solid phase reduced sulfur measurements and pore water sulfate profiles yield results that are consistent. Depth integrated sulfate reduction rates for modern sediments in San Pedro, Santa Catalina, and San Nicolas Basins are, 11.4, 6.3, and 6.3–8.8 (μmol cm‐2yr‐1), respectively. Measurements of solid phase iron species indicate that surficial sediments are enriched in easily‐reducible ferric oxyhydroxides. The enrichment is maintained by a combination of oxidation of Fe2+ diffusing upward from underlying anoxic sediments, as well as input of fresh sediment enriched in ferric oxyhydroxides. The three primary sources for iron converted to pyrite and the sequence in which they are utilized are: ferric oxyhydroxides, magnetite and other crystalline oxides, and “exchangeable” iron in phyllosilicates. The majority (50–80%) of the iron converted to pyrite is from the silicates, and budgetary calculations indicate the amount of iron released from San Pedro basin silicates agrees within 35% with the amount of magnesium removed from pore water to solid phases. Fe2+ is enriched in near‐surface pore waters because rates of dissolved iron production by oxyhydroxide reduction exceed rates of sulfate reduction and pyrite formation. At depth, pore waters are sulfidic because rates of sulfate reduction exceed rates of iron release from silicates. Sulfide produced at depth diffuses upward until it reaches sediments with available iron, causing a step‐like increase in solid phase sulfur concentration. Over 90% of the magnetite present in surficial sediments is dissolved at depth due to reaction with H2S. A model is developed to predict the depth at which magnetite dissolution should occur, based on sulfate reduction rates and the flux of ferric oxyhydroxides. The results of this model predict the onset of dissolution at depths of 5–40 cm in different basins and agree well with the observed depths of magnetite dissolution.

Adaptability of marine teleost renal inorganic sulfate excretion: evidence for glucocorticoid involvement.

The marine winter flounder, Pseudopleuronectes americanus, ingests seawater (SW) and excretes most of the resultant divalent ion load by renal tubule secretion. The site of secretion is generally thought to be the proximal tubule. Although sulfate clearance ratios (C ratio-sulfate clearance/polyethylene glycol clearance) have been reported as high as 12 in SW-acclimated animals, the present study shows that net secretion ceases after acclimation to 10% SW (SO4 free). Intravenous infusion of sulfate into the latter induced net secretion within 5 h (C ratio increased from 0.83 +/- 0.23 to 2.12 +/- 0.33). Increased sulfate secretion coincided with an increase in the magnitude of HCO3-SO4 exchange in brush-border membrane vesicles (BBMV) isolated from flounder renal tubules. Treatment of 10% SW-acclimated flounder with 60 micrograms dexamethasone/100 g body wt also caused an increase in the HCO3-SO4 exchange in BBMV to a level comparable to that of BBMV from SW fish. The glucocorticoid effect was further tested in flounder renal tubule primary monolayer cultures. Measurement of unidirectional sulfate fluxes (J) showed that in the presence of cortisol net secretion (serosal-to-mucosal flux) dominated (Js----m = 111 +/- 6.5, Jm----s = 9 +/- 4.3, Jnet = 102 +/- 2.2 nmol.h-1.cm-2). Removal of cortisol from the culture medium significantly reduced net sulfate secretion to one-third of control (Js----m = 39 +/- 13.9, Jm----s = 6 +/- 1.0, Jnet = 33 +/- 14.1 nmol.h-1.cm-2).

The regulation of sulphate uptake following growth of Pisum sativum L. seedlings in S nutrient limiting conditions. Interaction between nitrate and sulphate transport

The characteristics of sulphate transport into Pisum sativum L. Feltham First seedlings which had been grown in the presence and absence of a sulphur source ( ± S ) has been investigated. Short term (10 min) 35SO 4 2− influx into seedlings appeared to obey Michaelis-Menten kinetics over the concentration range 0–0.4 mol m −3 with apparent K m = 85 mmol m −3 and V max = 248 nmol g −1 fresh weight (FW) h −1 for the +S treatment and apparent K m = 70 mmol m −3 , and V max = 2500 nmol g −1 FW h −1 for the −S treatment. Biphasic kinetics were obvious in the former case (+S) only. 35SO 4 2− influx into the latter (−S) seedlings was inhibited by external nitrate in the influx medium ( K i = 0.24 mol m −3 ), but seedlings grown in +S conditions were insensitive to nitrate. The stimulation of sulphate influx following growth of the seedlings in −S conditions was confirmed from experiments which measured 35SO 4 2− efflux after loading in 35SO 4 2. Estimates of half times of filling of the cell wall and cytoplasmic phases were 2.5 min and 13–16 min, respectively. The rate of net sulphate uptake ( J) into seedlings grown in −S conditions rapidly declined over the first hour of resuply of sulphate, while SO 4 2− influx ( I) increased over the same time, and then gradually declined over the next 2-h. The results are interpreted in terms of a time dependent increase in rate of 35SO 4 2− efflux ( E). Nitrate stimulated E, but only from seedlings previously grown in −S conditions. The rate of substrate cycling defined as E/I increased following S deprivation.

Sulfate Enrichments in Estuarine Waters of North Carolina

Sulfate concentrations in the mesohaline surface waters of the Pamlico and Neuse River estuaries were enriched, relative to the conservative seawater ratio with Cl− (0.0517 (M)), by 5 to 43% between late winter and early summer. During this period, sulfate concentrations increased to a maximum excess of 3.5 mM in the bottom waters (0.5 m deep) through intermittent periods of both very low river flow and bottom water anoxia. The calculated net sulfate production rate for this period was 18 mmol per m2 per d in the bottom waters. By late summer, the excess sulfate (an average of 12 mol per m2) had been removed from the water column, presumably due to SO4 −2 reduction in anoxic bottom waters and sediments. Qualitative laboratory experiments with slurries of mud exposed to excess O2 and treated with inhibitors of cytochrome oxidase showed that it was possible to produce the excess SO4 −2 under these conditions via biochemical (not chemical) oxidation of pyride that occurs within the top 5 cm of mud (ca. 100 mmol pyrite-S per g dry mud). Whether the in situ substrate for SO4 −2 production was pyrite, S0, or S+2, is unknown, but the predominance of aqua regia extractable pyrite-Fe and the accumulation of excess SO4 −2 in slurries with insufficient other sources of oxidizable S, indicates that it may be an important substrate for biochemical production of SO4 −2.

Proton-sulfate cotransport: external proton activation of sulfate influx into human red blood cells.

Sulfate influx into human red blood cells was measured at 0 and 22 degrees C at several fixed external pH values between 3 and 10. These cells had normal internal pH and chloride concentrations so that sulfate influx was not limited by the efflux half-cycle reactions. The flux was a Michaelis-Menten function of sulfate concentration at each pH with K1/2SO4 = 4-10 mM. External protons activated influx 100-fold at a single site with a pK = 5.9 at 22 degrees C and 5.5 at 0 degrees C. This pK is similar to the value 5.99 +/- 0.3 for external proton binding to the sulfate-loaded transporter at 0 degrees C (J. Gen. Physiol. 79: 87-114, 1982). The flux was stilbene sensitive even in valinomycin-treated cells and was independent of membrane potential. This proton-activated influx appears to be proton-sulfate cotransport. At high pH there was a proton-independent flux that was membrane potential and stilbene sensitive. This proton-insensitive flux appears to be SO4(2-)/Cl- exchange or net sulfate influx. The sulfate influx over the entire pH range may be described in terms of an equation for the sum of the influxes through these two pathways on band 3.

Sulfate transport in rabbit proximal convoluted tubules: presence of anion exchange.

These studies describe the characteristics of sulfate transport in proximal convoluted tubules from rabbit kidney. We measured absorptive and secretory fluxes of sulfate in isolated tubular segments perfused and bathed with fluids containing sulfate concentrations of 0.2-10 mM. At 2.0 mM, the sulfate flux in the absorptive direction averaged 4.76 +/- 0.50 and the secretory flux was 3.08 +/- 0.31 pmol . mm-1 . min-1. Ouabain 10(-5) M decreased each to approx. 1.15 pmol . mm-1 . min-1. Kinetic analysis of each unidirectional sulfate flux demonstrated saturation with increasing sulfate concentrations. Thiosulfate 2 mM in the bath inhibited both absorptive and secretory sulfate fluxes; thiosulfate in the perfusate inhibited only the absorptive flux. Similar results were obtained with 10(-6) M SITS in either bath or perfusate. Phosphate had no effect on sulfate transport. Each unidirectional sulfate flux was influenced by the sulfate concentration in the solution on the opposite side in a pattern consistent with the presence of an anion exchange mechanism. Anion exchange transport persisted at 22 degrees C when net sulfate transport was abolished. The data indicate that sulfate transport in the proximal convoluted tubule is bidirectional, independent of phosphate transport, and occurs via two forms of facilitated transport, one of which is an anion exchange mechanism.

EFFECTS OF ADDED NITROGEN ON THE NET MINERALIZATION OF SOIL SULPHUR FROM TWO SOILS DURING INCUBATION

Net mineralizable sulphate, extractable with 0.15% CaCl2, was measured on two soils incubated with a variety of nitrogen sources and rates. Significant effects of source, rate and of interactions were observed, but the responses of the two soils were quite dissimilar. Both net mineralization and net immobilization were encountered.

The relationship between anion exchange and net anion flow across the human red blood cell membrane.

The conductive (net) anion permeability of human red blood cells was determined from net KCl or K2SO4 effluxes into low K+ media at high valinomycin concentrations, conditions under which the salt efflux is limited primarily by the net anion permeability. Disulfonic stilbenes, inhibitors of anion exchange, also inhibited KCl or K2SO4 efflux under these conditions, but were less effective at lower valinomycin concentrations where K+ permeability is the primary limiting factor. Various concentrations of 4,4'-diisothiocyanostilbene-2,2'-disulfonate (DIDS) had similar inhibitory effects on net and exchange sulfate fluxes, both of which were almost completely DIDS sensitive. In the case of Cl-, a high correlation was also found between inhibition of net and exchange fluxes, but in this case about 35% of the net flux was insensitive to DIDS. The net and exchange transport processes differed strikingly in their anion selectivity. Net chloride permeability was only four times as high as net sulfate permeability, whereas chloride exchange is over 10,000 times faster than sulfate exchange. Net OH-permeability, determined by an analogous method, was over four orders of magnitude larger than that of Cl-, but was also sensitive to DIDS. These data and others are discussed in terms of the possibility that a common element may be involved in both net and exchange anion transport.