Wood-based biochar as an excellent activator of peroxydisulfate for Acid Orange 7 decolorization.

Wood-based biochar, as a metal-free heterogeneous activator of peroxydisulfate (PDS), was successfully prepared by pyrolysis of polar sawdust for efficient removal of Acid Orange 7 (AO7). The results demonstrate PDS could be effectively activated by wood-based biochar, and AO7 was rapidly eliminated in a wide range of pH value (3.0-10.0) with AO7 removal achieved ≥ 99.3% after 14 min reaction. The dominant reactive species in the biochar/PDS system were verified via radical quenching tests and electron paramagnetic resonance (EPR) technique. It is speculated that sulfate radicals (SO4•-) and hydroxyl radicals (•OH) were formed on the surface of biochar. Based on the results of X-ray photoelectron spectroscopy (XPS), π-electron density and oxygen-containing functional groups (especially C-OH) on biochar surface were active centers for the catalytic reaction. Recycle experiments of biochar for 4 runs were carried out and the regeneration method of the catalyst was also studied.

Calcium ameliorates the toxicity of sulfate salinity in Brassica rapa.

Salinity stress in Brassica, often only associated with osmotic effects and the toxicity of Na+, was more severe when applied as Na2SO4 than as NaCl, indicating that SO42- ions had toxic effects as well. Application of 10 mM calcium in the form of CaCl2 in the growth medium of plants only slightly ameliorated growth impairment by NaCl and KCl, but almost completely prevented negative effects of Na2SO4 and K2SO4 on plant biomass production. This effect was calcium specific, as MgCl2 ameliorated sulfate toxicity to a much lower extent. This sulfate toxicity coincided with a strong decrease in the plant content of calcium and manganese upon sulfate salinity. Application of CaCl2 largely alleviated this decrease, however, it did not prevent the higher tissue concentration of sulfate. CaCl2 prevented the increase in organic sulfur compounds presumably by reducing of relative gene expression of ATP-sulfurylase (ATPS) and adenosine 5'-phosphosulfate reductase (APR) indicating a possible regulation of sulfate assimilation by calcium. The upregulation of the genes encoding for Group 4 sulfate transporters (Sultr4;1 and 4;2) upon sulfate salinity, was absent in the presence of CaCl2. Therefore, additional calcium may facilitate an increased vacuolar capacity for sulfate accumulation.

Chemical inhibition of sulfation accelerates neural differentiation of mouse embryonic stem cells and human induced pluripotent stem cells.

Pluripotency of embryonic stem cells (ESCs) is maintained by the balancing of several signaling pathways, such as Wnt, BMP, and FGF, and differentiation of ESCs into a specific lineage is induced by the disruption of this balance. Sulfated glycans are considered to play important roles in lineage choice of ESC differentiation by regulating several signalings. We examined whether reduction of sulfation by treatment with the chemical inhibitor chlorate can affect differentiation of ESCs. Chlorate treatment inhibited mesodermal differentiation of mouse ESCs, and then induced ectodermal differentiation and accelerated further neural differentiation. This could be explained by the finding that several signaling pathways involved in the induction of mesodermal differentiation (Wnt, BMP, and FGF) or inhibition of neural differentiation (Wnt and BMP) were inhibited in chlorate-treated embryoid bodies, presumably due to reduced sulfation on heparan sulfate and chondroitin sulfate. Furthermore, neural differentiation of human induced pluripotent stem cells (hiPSCs) was also accelerated by chlorate treatment. We propose that chlorate could be used to induce efficient neural differentiation of hiPSCs instead of specific signaling inhibitors, such as Noggin.

Metal sulfate-mediated induction of pathogenic genes and repression by phenyl butyl nitrone and Feralex-G.

Neurotoxic metal-induced oxidative damage to nervous tissue has been implicated in several progressive neurodegenerative disorders including Alzheimer's disease. In this study, using human brain cells in primary culture, the quenching of metal sulfate-induced reactive oxygen species (ROS) and ROS-sensitive gene expression was studied using the antioxidants ascorbate, folic acid, phenyl butyl nitrone and the chelators desferrioxamine and Feralex-G. Antioxidants ascorbate, folic acid, phenyl butyl nitrone, desferrioxamine or Feralex-G were found to quench ROS and cPLA2 and COX-2 gene induction to various degrees, and a synergism was observed when certain combinations of them were used. These findings support the idea that specific antioxidants and metal ion chelators when used together can effectively and synergistically quench ROS-mediated induction of pathogenic gene expression.

Regulation of the atherogenic properties of vascular smooth muscle proteoglycans by oral anti-hyperglycemic agents.

The present study aimed to investigate the actions of several classes of oral hypoglycemic agents [e.g., sulfonylureas (SUs), biguanides (BGs) and thiazolidinediones (TZDs)] in an in vitro model of lipid binding based on the "response to retention" hypothesis of atherogenesis. The incorporation of [(35)S]-SO(4) into proteoglycans synthesized by human vascular smooth muscle cells (VSMCs) was assessed by cetylpyridinium chloride (CPC) precipitation method, proteoglycan electrophoretic mobility was evaluated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and binding to low-density lipoprotein (LDL) was assessed by gel mobility shift assay (GMSA). The SUs evaluated showed no effect on [(35)S]-SO(4) incorporation into proteoglycans. Only one BG, phenformin, caused a concentration-related inhibition of proteoglycan synthesis under basal conditions and in the presence of transforming growth factor-beta1 (TGF-beta1), caused by an inhibition of proteoglycan core protein synthesis secondary to a reduction in total protein synthesis. However, neither metformin nor phenformin (30-300 micromol/l) had any effect on the electrophoretic mobility of proteoglycans. The TZDs--troglitazone (TRO), rosiglitazone (ROS), and pioglitazone (PIO) (10, 30, and 30 micromol/l, respectively)--inhibited proteoglycan biosynthesis and stimulated total proteoglycan core protein synthesis, while TRO alone inhibited overall protein synthesis. All three TZDs moderately reduced the electrophoretic mobility of synthesized proteoglycans assessed by SDS-PAGE, reduced the sizes of cleaved glycosaminoglycan (GAG) chains assessed by size exclusion chromatography, and significantly reduced binding to LDL. The data indicate that TZDs show anti-atherogenic actions through the modification of proteoglycan structure, leading to a possible reduction in lipid retention in the vessel wall.

Sulfation is required for bone morphogenetic protein 2-dependent Id1 induction.

Different reports have suggested the dependence of bone morphogenetic protein (BMP) activity on the sulfated glycosaminoglycan (GAG) chains found in proteoglycans. However, the requirement of sulfated molecules in early BMP-2-signaling responses has not been established. We have used sodium chlorate to inhibit sulfation in C2C12 cells and have analyzed BMP-2 induction of Id1. We show here that sulfation inhibition strongly decreases the specific and early induction of Id1 at the transcriptional level. This effect is not reverted by the addition of extracellular components, such as GAGs or extracellular matrix (ECM). The inhibition of GAG incorporation into proteoglycans, or their removal by GAG lyases, does not mimic the negative effect on Id1 expression, while sulfation inhibition also represses the Id1-induction exerted by a constitutively active form of the BMP receptor, suggesting that BMP-2-mediated Id1 induction has an intracellular requirement for sulfated molecules.

Heparan sulfate proteoglycans modulate monocyte migration across cerebral endothelium.

Heparan sulfate proteoglycans (HSPGs) are known to participate in a wide range of biological events, including cellular trafficking. In this study we report that in situ cerebral blood vessels highly express HSPGs. Of the syndecan family, syndecan-2 is highly expressed on virtually all brain vessels and syndecan-1 and -3 are only present on larger blood vessels. These endothelial HSPGs have a functional role in monocyte diapedesis across brain endothelium, as assessed in our in vitro adhesion and migration assays. Our data indicate that heparin prevents monocyte adhesion to brain endothelium by interacting solely with the monocyte. Transendothelial migration of monocytes can be prevented by preincubation of brain endothelium with heparin by enzymatic removal of heparan sulphate side chains or by inhibition of cellular sulfation. Blocking of G-protein-dependent signaling in the monocytes prevented monocyte adhesion and migration to similar extent, suggesting that G-dependent signaling may be involved in HSPG-mediated monocyte adhesion and transendothelial migration. Our data demonstrate that brain endothelial HSPGs have a modulatory role in the transendothelial migration of monocytes in a direct and indirect fashion and may therefore contribute to the formation of neuroinflammatory lesions.

Binding properties of the stilbene disulfonate sites on human erythrocyte AE1: kinetic, thermodynamic, and solid state deuterium NMR analyses.

A novel stilbene disulfonate, 4-trimethylammonium-4'-isothiocyanostilbene-2,2'-disulfonic acid (TIDS), has been chemically synthesized, and the interaction of this probe with human erythrocyte anion exchanger (AE1) was characterized. Covalent labeling of intact erythrocytes by [N(+)((14)CH(3))(3)]TIDS revealed that specific modification of AE1 was achieved only after removal of other ligand binding sites by external trypsinization. Following proteolysis, (1.2 +/- 0.4) x 10(6) TIDS binding sites per erythrocyte could be blocked by prior treatment with 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS), a highly specific inhibitor of AE1. Inhibition of sulfate equilibrium exchange by TIDS in whole cells was described by a Hill coefficient of 1.10 +/- 0.06, which reduced to 0.51 +/- 0.01 following external trypsinization. The negative cooperativity of TIDS binding following external trypsinization suggests that trypsin-sensitive proteins modulate allosteric coupling between AE1 monomers. Thermodynamic analysis revealed that TIDS binding induces smaller conformational changes in AE1 than is observed following DIDS binding. The similar inhibitory potencies of both TIDS (IC(50) = 0.71 +/- 0.48 microM) and DIDS (IC(50) = 0.2 microM) imply that there is no correlation between the ability of stilbene disulfonates to arrest anion exchange function and the magnitude of ligand-induced conformational changes in AE1. Solid state (2)H NMR analysis of a [N(+)(CD(3))(3)]TIDS-AE1 complex in both unoriented and macroscopically oriented membranes revealed that large amplitude "wobbling" motions describe ligand dynamics. The data are consistent with a model where TIDS bound to AE1 is located exofacially in contact with the bulk aqueous phase.

Metabolism and cytotoxicity of chlorpropham (CIPC) and its essential metabolites in isolated rat hepatocytes during a partial inhibition of sulphation and glucuronidation reactions: a comparative study.

The changes in metabolism and cytotoxicity of chlorpropham (CIPC) and its major metabolites, 4-hydroxychlorpropham (4-OH CIPC), 3-chloroaniline, and 3-chloroacetanilide were investigated in isolated rat hepatocyte suspensions after a partial inhibition of sulphation and glucuronidation and the two reactions combined in an attempt to assess the part of each of them in the enhanced CIPC toxicity observed in vivo after D-galactosamine treatment. With sulphation and glucuronidation effective, CIPC has a cytolytic effect and reduces intracellular ATP and K+ level while 4-OH CIPC has a weak cytolytic effect but modifies ATP and K+ level in a greater extent than CIPC. Inhibition of sulphation does not affect the cytotoxicity of CIPC or 4-OH CIPC because there is a compensatory increase in the amount of 4-OH CIPC glucuronide formed and the level of free 4-OH CIPC always remain low. In contrast, when incubations are carried out with either CIPC or 4-OH CIPC, the presence of D-galactosamine leads to a decrease of glucuronide and sulphate conjugates accompanied, respectively, by a 3.6-fold and 6. 9-fold increase of the free 4-OH CIPC level in the culture medium. This alteration of the metabolism is followed by a marked reduction of ATP synthesis with a concomitant modification of cell permeability. The cytolytic effect is due to CIPC itself, whereas the effect on energy supply was attributed to free 4-OH CIPC. The results demonstrate a combined effect of free 4-OH CIPC and D-galactosamine on intracellular ATP level that could account for the partial inhibition of sulphation. This change in the CIPC metabolism could explain the increased CIPC toxicity observed in vivo after D-galactosamine pretreatment.

Effects of brefeldin A on galactosphingolipid synthesis in an immortalized Schwann cell line: evidence for different intracellular locations of galactosylceramide sulfotransferase and ceramide galactosyltransferase activities.

Brefeldin A (BFA) has been used extensively to study the intracellular transport and processing of proteins and sphingolipids because of its dramatic alteration of the structural and functional organization of the Golgi. We have examined the effect of BFA on the synthesis of galactosylceramide sulfate (SGalCer) and its immediate precursor galactosylceramide (GalCer) in an immortalized Schwann cell line (S16) to determine the intracellular sites of synthesis of these two related glycolipids. During a 6-h labeling period, a dose-dependent inhibition of [35S]sulfate incorporation into SGalCer was observed with 95% inhibition occurring at 0.5 microgram/ml BFA. Labeling of newly synthesized galactosphingolipids with [3H]-palmitic acid for 6 h in the presence of BFA resulted in increased incorporation of label into GalCer containing nonhydroxy fatty acids (NFA-GalCer) to 162% of control values, whereas labeling of GalCer containing 2-hydroxy fatty acids (HFA-GalCer) was reduced to 63% of control. After 24 h, these values were at 366 and 91%, respectively. These results indicate that at least some of the HFA-GalCer was initially synthesized at a location distal to the BFA block and separate from the site of NFA-GalCer synthesis. Examination of [3H]palmitic acid incorporation into free ceramides showed an increase of 133 and 161% for hydroxy and nonhydroxy fatty acid ceramides, respectively, in cells treated for 6 h with BFA in comparison with levels found in untreated control cells, indicating that BFA did not block fatty acid 2-hydroxylation or the formation of HFA ceramide.(ABSTRACT TRUNCATED AT 250 WORDS)

Changes in protein sulfation in human erythroleukemia (HEL), CHRF-288-11, and K562 cells following treatment with dimethyl sulfoxide or phorbol 12-myristate 13-acetate.

This study has demonstrated that three hematopoietic tumor cell lines with megakaryocytic characteristics, HEL, CHRF-288-11, and K562, synthesize a number of sulfated proteins. The major HEL sulfated proteins were a doublet at 88 and 92 kDa and several closely spaced bands between 125 and 160 kDa and more acidic proteins of 210 kDa. Treatment with dimethylsulfoxide (DMSO) for 24 h almost completely inhibited labeling of sulfated proteins, and up to 48 h, labeling was found almost entirely in a band at 125 kDa. Treatment with phorbol 12-myristate 13-acetate (PMA) nearly eliminated labeling of the 88- and 92-kDa bands and resulted in the appearance of a large amount of labeling between 96 and 108 kDa. Sulfated proteins of 135 and 210 kDa were immunoprecipitated by an antibody against platelet GP Ib. A 130-kDa protein was immunoprecipitated by an antibody against the beta-1 integrin subunit. The major proteins labeled in CHRF cells were at 68, 90, 98, 125, and 148 kDa. Treatment with PMA greatly reduced the labeling of the 148-kDa band, eliminated the labeling of the 68-kDa band, and markedly enhanced labeling of the 92-kDa region. The major proteins labeled in K562 cells were at 110, 120-130, and 145 kDa. PMA reduced the labeling of the 110- and 145-kDa proteins and extensively increased labeling of bands at 120-130, 78, and 84 kDa, and DMSO caused decreased labeling of the 120- to 130-kDa proteins. This is the first demonstration of sulfation of specific proteins in hematopoietic cell lines and of the alteration of sulfation of specific proteins in any cells in response to treatment with differentiation-inducing agents. We hypothesize that changes in sulfation of proteins may be relevant to the maturation or malignant growth of megakaryocytic cells.

Comparative aspects of utilization of sulfonate and other sulfur sources by Escherichia coli K12.

Selected biochemical features of sulfonate assimilation in Escherichia coli K-12 were studied in detail. Competition between sulfonate-sulfur and sulfur sources with different oxidation states, such as cysteine, sulfite and sulfate, was examined. The ability of the enzyme sulfite reductase to attack the C-S linkage of sulfonates was directly examined. Intact cells formed sulfite from sulfonate-sulfur. In cysteine-grown cells, when cysteine was present with either cysteate or sulfate, assimilation of both of the more oxidized sulfur sources was substantially inhibited. In contrast, none of three sulfonates had a competitive effect on sulfate assimilation. In studies of competition between different sulfonates, the presence of taurine resulted in a decrease in cysteate uptake by one-half, while in the presence of isethionate, cysteate uptake was almost completely inhibited. In sulfite-grown cells, sulfonates had no competitive effect on sulfite utilization. An E. coli mutant lacking sulfite reductase and unable to utilize isethionate as the sole source of sulfur formed significant amounts of sulfite from isethionate. In cell extracts, sulfite reductase itself did not utilize sulfonate-sulfur as an electron acceptor. These findings indicate that sulfonate utilization may share some intermediates (e.g., sulfite) and regulatory features (repression by cysteine) of the assimilatory sulfate reductive pathway, but sulfonates do not exert regulatory effects on sulfate utilization. Other results suggest that unrecognized aspects of sulfonate metabolism, such as specific transport mechanisms for sulfonates and different regulatory features, may exist.

The inhibitory effects of amrinone on isolated rat uterus.

Amrinone, a new cardiotonic drug, has received attention as a better therapeutic agent than the cardiac glycosides in the treatment of congestive heart failure. In this study, the effects of amrinone on isolated rat uterus and its probable mechanism of action were investigated. At two different concentrations (0.1 and 0.5 mM), the inhibitory effects of amrinone on the spontaneous contractions of rat uterus were noted. In addition, amrinone (0.5 mM) was found to inhibit the tonic contractions induced by potassium sulphate (K2SO4)-Ringer solution (91.74%) and calcium chloride (CaCl2) (93.04%). These inhibitory effects were compared with regulators of the phosphodiesterase enzyme (PDE). It was concluded that amrinone could behave as a calcium antagonist and PDE inhibitor.

Hepatic triiodothyronine sulfation and its regulation by growth hormone and triiodothyronine in rats.

The regulatory mechanism of cytosolic sulfation of T3 has been studied in rat liver. Sulfation of T3 is sexually differentiated in adult rats of Sprague-Dawley (SD), Fisher 344, and ACI strains. In SD strain, the male animals showed 4 times higher sulfating activity than did the females. The specific activity was decreased by hypophysectomy of male adult rats, but was not affected in the females. Thus, the sex-difference was abolished in the hypophysectomized condition. Supplement of human GH intermittently twice daily for 7 days, to mimic the male secretory pattern, increased T3 sulfating activity in both sexes of hypophysectomized rats, whereas continuous infusion to mimic a female secretory pattern had no appreciable effect. Cytosolic sulfation of T3 was decreased by 25 to 30% by thyroidectomy or propylthiouracil treatment of male adult rats, and was restored by the supplementation of T3 (50 micrograms/kg daily for 7 days) to thyroidectomized rats. Administration of T3 in hypophysectomized rats almost completely restored the sulfating activity in the males and increased the activity in the females. Cytosolic T3 sulfation was inhibited by the addition of known inhibitors of phenol sulfotransferase, pentachlorophenol or 2,6-dichloro-4-nitrophenol. These results indicate a role of pituitary GH in hepatic sulfation of thyroid hormones in rats. The data obtained also raise the possibility that GH may modify the effect of thyroid hormones on the pituitary by a feed-back mechanism through changing the level of a sex-dominant phenol sulfotransferase(s) in rat livers. T3 was also sulfated in hepatic cytosols of mouse, hamster, rabbit, dog, monkey, and human.(ABSTRACT TRUNCATED AT 250 WORDS)

Mitogenic and dentin-inductive effects of crude bone morphogenetic protein from bone and dentin in primary adult pulp cell culture.

The effects of crude bone morphogenetic protein (BMP) derived from bone and dentin matrix on proliferation, production of extracellular matrix, and biologic function of the pulp cell were examined in the primary cell culture from permanent dental pulp. BMP from bone and from dentin matrix stimulated iodine 125-deoxyuridine incorporation in the absence of 10% calf serum. They increased sulfur 35-sulfate incorporation in proliferating stage and had no effects in stationary stage of culture. Alkaline phosphatase activities were inhibited in proliferating, stationary, and multilayered stages of culture. Osteocalcin synthesis was increased in culture treated with BMP from day 2 to day 10. These findings suggest that crude BMP might have mitogenic activity and some role in regulation of differentiation of pulp cells into odontoblasts.

Electrogenic H(+)-regulated sulfate-chloride exchange in lobster hepatopancreatic brush-border membrane vesicles.

Transport of [35S]sulfate by brush-border membrane vesicles (BBMV) of lobster (Homarus americanus) hepatopancreas was stimulated by an outwardly directed chloride gradient. In contrast, sulfate uptake was not enhanced by inwardly directed Na+ or K+ transmembrane gradients. An inside-positive membrane potential (valinomycin and K+) stimulated SO4(2-)-Cl- exchange, whereas an inside-negative membrane potential was inhibitory. Sulfate-sulfate exchange was not affected by alterations of transmembrane potential. An inwardly directed proton gradient, or the presence of low bilateral pH, enhanced SO4(2-)-Cl- exchange, but the H+ gradient alone did not stimulate sulfate uptake in chloride-equilibrated BBMV or in vesicles lacking internal Cl-. The stilbenes 4-acetamido-4'-isothiocyanostilbene-2,2'-disulfonic acid (SITS) and 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS) strongly inhibited SO4(2-)-Cl- exchange. Sulfate influx occurred by a combination of carrier-mediated transfer, exhibiting Michaelis-Menten kinetics, and nonsaturable "apparent diffusion." 36Cl- influx into sulfate-loaded BBMV was stimulated by an inside-negative transmembrane potential compared with short-circuited vesicles. These results suggest that sulfate-chloride exchange in hepatopancreatic BBMV occurred by an electrogenic carrier mechanism exhibiting a 1:1 flux ratio that was modulated by internal and external H(+)-sensitive regulatory sites. The role of this antiport process in anion secretion is discussed.

Occurrence of sulfate in an asparagine-linked complex oligosaccharide of chicken adipose lipoprotein lipase.

After adipocytes were labeled with Na2[35SO4], immunoadsorbed with immobilized antilipoprotein lipase, and subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and fluorography, a labeled band was identified at 59,700 daltons, the molecular mass of chicken lipoprotein lipase (LPL). Excess unlabeled LPL prevented the immunoadsorption of this labeled species, hence the labeled species was determined to be LPL. Digestion of LPL with endo-beta-N-acetylglucosaminidase H (Endo H) caused a shift in mobility of LPL in SDS-PAGE with no loss of radioactivity, whereas digestion with glycopeptidase F resulted in removal of 99% of the radioactivity. Adipocytes cultured with Trans35S-label and tunicamycin produced an LPL species of 52,000 daltons, but tunicamycin abolished the incorporation of 35SO4 into LPL. This established that 35SO4 was incorporated into an N-linked oligosaccharide of LPL. Endo H digestion of pulse-chase labeled LPL revealed the presence of two complex and one high mannose-type N-linked oligosaccharides. A single 35SO4-labeled tryptic peptide was isolated by reverse phase chromatography. The amino acid sequence of the peptide established that the 35SO4 oligosaccharide is conjugated at Asn-45. Behavior of the 35SO4-labeled oligosaccharide on concanavalin A-agarose, sequential exoglycosidase digestion, and chemical analysis of the 35SO4 oligosaccharide confirms that this moiety is of the complex type. Sequential exoglycosidase digestion, thin layer chromatography of the released monosaccharides, and the use of glycosylation inhibitors established that the sulfated sugar is a core N-acetylglucosamine (GlcNAc). The data show that chicken LPL contains two complex and one high mannose N-linked oligosaccharides and that 35SO4 is incorporated into LPL on a GlcNAc residue of a complex oligosaccharide located at Asn-45.

[Antagonism of Zn2+ on nootropic action of piracetam in mice].

In mouse step-down test, the memory impairments of acquisition, consolidation and recognition were induced by anisodine, chloramphenical and ethanol, respectively. Piracetam 100 mg/(kg.d) ip for 5 d improved the anisodine-induced impairment of learning. ZnSO4 5 mg/(kg.d) po for 5 d did not improve the 3 impairments. Memory impairments were enhanced by a combined administration of ZnSO4 and piracetam in these 3 models. These results were confirmed by Y-maze method in normal mice.

A study of sulphate transport by lactating rat mammary tissue slices: evidence for anion exchange.

1. The efflux of radiolabelled sulphate from lactating rat mammary tissue slices has been studied. Sulphate efflux was found to be time- and temperature-dependent. 2. 4,4'-Diisothiocyanostilbene-2,2'-disulphonate (DIDS) inhibited a portion of sulphate release, whereas bumetanide was without effect. 3. The anions chloride, iodide and sulphate trans-stimulated sulphate efflux when added to the incubation medium. The increase in the efflux rate of sulphate found with chloride could be markedly inhibited by DIDS. Thiocyanate, unlike the other anions tested, only had a small effect. 4. The results strongly suggest that there is an anion exchange mechanism in the mammary gland which can mediate the transport of sulphate. This transporter may be important for the metabolism of sulphate by the mammary gland and may also help determine milk anion concentrations.

The effect of beta 2-glycoprotein I on the dextran sulfate and sulfatide activation of the contact system (Hageman factor system) in the blood coagulation.

1. beta 2-Glycoprotein I, inhibits the initiation of the contact system in plasma accomplished by dextran sulfate. 2. The dextran sulfate induced activation could be inhibited both when dextran sulfate was preincubated with beta 2-glycoprotein I and when the amount of beta 2-glycoprotein I in plasma was increased. 3. The concentration of beta 2-glycoprotein I at which an inhibitory effect could be registered was dependent upon the concentration of negatively charged groups on the surface. Calculation of the molar ratios between beta 2-glycoprotein I and sulfate residues in dextran sulfate showed that beta 2-glycoprotein I had to be present in excess of a 1:1 stoichiometric ratio of the sulfate group in order to inhibit the activation. 4. beta 2-Glycoprotein I does not inhibit the initiation of the contact system in plasma accomplished by sulfatide, unless the sulfatide has been preincubated with beta 2-glycoprotein I.