Effect of sodium hydrosulphite treatment on the quality of non-centrifugal sugar: Jaggery.

Jaggery is a non-centrifugal sweetener produced by thermo-chemical treatments of sugarcane juice. The traditional practices involved in making jaggery are usually tailored using chemicals to meet consumer requirements. Sodium hydrosulphite (hydros) is a commonly employed chemical in the jaggery industry to improve its colour. This article presents a comparative study of jaggery made with and without hydros treatments. The differences in properties, such as sorption behaviour, colour, polyphenols, flavonoids, minerals, and sulphur dioxide content, were measured. Hydros-treated jaggery was found to be brighter in colour with a lower browning index by 5-10. SO2 content of hydros-treated jaggery was >70 ppm, while minerals, polyphenols, and flavonoids were less abundant compared to control jaggery, thereby compromising overall quality. Based on the experiments carried out, the optimum treatment of hydros can be employed to satisfy consumer demand while producing an acceptable quality of jaggery that conforms to norms.

Photorespiration participates in the assimilation of acetate in Chlorella sorokiniana under high light.

The development of microalgae on an industrial scale largely depends on the economic feasibility of mass production. High light induces productive suspensions during cultivation in a tubular photobioreactor. Herein, we report that high light, which inhibited the growth of Chlorella sorokiniana under autotrophic conditions, enhanced the growth of this alga in the presence of acetate. We compared pigments, proteomics and the metabolic flux ratio in C. sorokiniana cultivated under high light (HL) and under low light (LL) in the presence of acetate. Our results showed that high light induced the synthesis of xanthophyll and suppressed the synthesis of chlorophylls. Acetate in the medium was exhausted much more rapidly in HL than in LL. The data obtained from LC-MS/MS indicated that high light enhanced photorespiration, the Calvin cycle and the glyoxylate cycle of mixotrophic C. sorokiniana. The results of metabolic flux ratio analysis showed that the majority of the assimilated carbon derived from supplemented acetate, and photorespiratory glyoxylate could enter the glyoxylate cycle. Based on these data, we conclude that photorespiration provides glyoxylate to speed up the glyoxylate cycle, and releases acetate-derived CO2 for the Calvin cycle. Thus, photorespiration connects the glyoxylate cycle and the Calvin cycle, and participates in the assimilation of supplemented acetate in C. sorokiniana under high light.

EPR monitored redox titration of the cofactors of Saccharomyces cerevisiae Nar1.

Electron Paramagnetic Resonance (EPR) monitored redox titrations are a powerful method to determine the midpoint potential of cofactors in proteins and to identify and quantify the cofactors in their detectable redox state. The technique is complementary to direct electrochemistry (voltammetry) approaches, as it does not offer information on electron transfer rates, but does establish the identity and redox state of the cofactors in the protein under study. The technique is widely applicable to any protein containing an electron paramagnetic resonance (EPR) detectable cofactor. A typical titration requires 2 ml protein with a cofactor concentration in the range of 1-100 µM. The protein is titrated with a chemical reductant (sodium dithionite) or oxidant (potassium ferricyanide) in order to poise the sample at a certain potential. A platinum wire and a Ag/AgCl reference electrode are connected to a voltmeter to measure the potential of the protein solution A set of 13 different redox mediators is used to equilibrate between the redox cofactors of the protein and the electrodes. Samples are drawn at different potentials and the Electron Paramagnetic Resonance spectra, characteristic for the different redox cofactors in the protein, are measured. The plot of the signal intensity versus the sample potential is analyzed using the Nernst equation in order to determine the midpoint potential of the cofactor.

Characterization of colloidal Fe from soils using field-flow fractionation and Fe K-edge X-ray absorption spectroscopy.

Colloids may facilitate the transport of trace elements and nutrients like phosphate in soil. In this study, we characterized soil colloids (<0.45 µm), extracted from four agricultural soils by Na-bicarbonate and Na-pyrophosphate, by two complementary analytical techniques; asymmetric flow field-flow fractionation (AF4) and X-ray absorption spectroscopy (XAS). The combined results from AF4 and XAS show that colloidal Fe is present as (i) free Fe-(hydr)oxide nanoparticles, (ii) Fe-(hydr)oxides associated with clay minerals, and (iii) Fe in clay minerals. Free Fe-(hydr)oxide nanoparticles, which can be as small as 2-5 nm, are extracted with Na-pyrophosphate but not with Na-bicarbonate, except for one soil. In contrast, Fe-(hydr)oxides associated with clay minerals are dispersed by both extractants. XAS results show that the speciation of Fe in the colloidal fractions closely resembles the speciation of Fe in the bulk soil, indicating that dispersion of colloidal Fe from the studied soils was rather unselective. In one Fe-rich soil, colloidal Fe was dominantly dispersed in the form of free Fe-(hydr)oxide nanoparticles. In the other three soils, dispersed Fe-(hydr)oxides were dominantly associated with clay minerals, suggesting that their dispersion as free nanoparticles was inhibited by strong attachment. However, in these soils, Fe-(hydr)oxides can be dispersed as oxide-clay associations and may as such facilitate the transport of trace elements.

[Study on protective effect of cerebrospinal fluid containing Qingxin Kaiqiao Fang on sodium dithionite-induced PC12 cell injury].

OBJECTIVE: To study the protective effect of cerebrospinal fluid containing Qingxin Kaiqiao Fang on sodium dithionite (Na2S2O4)-induced PC12 cell injury, in order to provide basis for clinical application of the prescription. METHOD: SD rats were orally administered with water decoction of Qingxin Kaiqiao Fang (7. 9 g . kg-1) once every 12 h, for a total of 7 times, in order to prepare cerebrospinal fluid containing Qingxin Kaiqiao Fang. The neurocyte injury model was established by adding Na2S2O4 with the final concentration of 8 m mol . L-1 into PC12 cells. With nimodipine (1 x 10(7)mol . L-1 ) as the positive control group, MTT method test was adopted to detect the impact of cerebrospinal fluid containing Qingxin Kaiqiao Fang on the activity of PC12 cells. The expression of Bax, Bel-2 and Caspase-3 mRNA was detected by RT-PCR. RESULT: The cerebrospinal fluid containing Qingxin Kaiqiao Fang groups showed a significantly higher activity in PC12 cells than the model group, with decrease in expressions of Bax mRNA and Caspase-3 mRNA and increase in expression of Bel-2 mRNA. There were significant differences compared with the model group (P< 0. 05,P <0. 01). CONCLUSION: Qingxin Kaiqiao Fang shows a notable protective effect on Na2S2 04-induced neurocyte injury.

Study on protective effect of cerebrospinal fluid containing Qingxin Kaiqiao Fang on sodium dithionite-induced PC12 cell injury / 中国中药杂志

<p><b>OBJECTIVE</b>To study the protective effect of cerebrospinal fluid containing Qingxin Kaiqiao Fang on sodium dithionite (Na2S2O4)-induced PC12 cell injury, in order to provide basis for clinical application of the prescription.</p><p><b>METHOD</b>SD rats were orally administered with water decoction of Qingxin Kaiqiao Fang (7. 9 g . kg-1) once every 12 h, for a total of 7 times, in order to prepare cerebrospinal fluid containing Qingxin Kaiqiao Fang. The neurocyte injury model was established by adding Na2S2O4 with the final concentration of 8 m mol . L-1 into PC12 cells. With nimodipine (1 x 10(7)mol . L-1 ) as the positive control group, MTT method test was adopted to detect the impact of cerebrospinal fluid containing Qingxin Kaiqiao Fang on the activity of PC12 cells. The expression of Bax, Bel-2 and Caspase-3 mRNA was detected by RT-PCR.</p><p><b>RESULT</b>The cerebrospinal fluid containing Qingxin Kaiqiao Fang groups showed a significantly higher activity in PC12 cells than the model group, with decrease in expressions of Bax mRNA and Caspase-3 mRNA and increase in expression of Bel-2 mRNA. There were significant differences compared with the model group (P< 0. 05,P <0. 01).</p><p><b>CONCLUSION</b>Qingxin Kaiqiao Fang shows a notable protective effect on Na2S2 04-induced neurocyte injury.</p>

Speciation of Al, Fe, and P in recent sediment from three lakes in Maine, USA.

Sequential extraction of sediments [Psenner R, Pucsko R. Die Fraktionierung organischer und anorganischer Phosphorverbindungen von Sedimenten. Arch Hydrobiol/Suppl 1988. 70(1): 111-155.] from short, (210)Pb-dated cores from three lakes in Maine USA demonstrates that sediment P is dominantly associated with the NaOH-extractable fraction (P-NaOH(25)) and less with the bicarbonate-dithionite extractable fraction (P-BD). The ratios (Al-NaOH(25))/(Fe-BD) and (Al-NaOH(25))/(P-NH(4)Cl+P-BD) for upper sediment for two oligo-mesotrophic lakes exceeded 3 and 25, the thresholds for preventing substantial release of P from sediments during hypolimnetic anoxia [Kopácek J, Borovec J, Hejzlar J, Ulrich K-U, Norton SA, Amirbahman A. Aluminum control of phosphorus sorption by lake sediments. Environ Sci Technol 2005a;39:8784-8789.]. Hypolimnetic water chemistry verifies this effect. The third lake, currently eutrophic, has values for the ratios that are below the thresholds and this lake has substantial release of P from recent sediment. The sediment characteristics remain relatively constant over the last 150+ years, indicating that the processes responsible for P retention have operated long before atmospheric acidification of watersheds might have influenced the flux of Al and Fe to the lake. In 2002, the pH of inlets and the lakes was generally between 6 and 8. Input to the lakes had high concentrations of acid-soluble particulate and dissolved Al, Fe, and P, and dissolved Al and Fe complexed with dissolved organic carbon (DOC). Lake water column and outlet Al, Fe, and P were typically 90-95% lower than inlet concentrations over a 12 month period. Photo-oxidation of Al-DOC and Fe-DOC in the lake, liberation of inorganic Al and Fe, precipitation of Al(OH)(3) and Fe(OH)(3), adsorption of P by the hydroxides, and sedimentation are responsible for the changes in water quality and long-term sediment characteristics.

Joziknipholones A and B: the first dimeric phenylanthraquinones, from the roots of Bulbine frutescens.

From the roots of the African plant Bulbine frutescens (Asphodelaceae), two unprecedented novel dimeric phenylanthraquinones, named joziknipholones A and B, possessing axial and centrochirality, were isolated, together with six known compounds. Structural elucidation of the new metabolites was achieved by spectroscopic and chiroptical methods, by reductive cleavage of the central bond between the monomeric phenylanthraquinone and -anthrone portions with sodium dithionite, and by quantum chemical CD calculations. Based on the recently revised absolute axial configuration of the parent phenylanthraquinones, knipholone and knipholone anthrone, the new dimers were attributed to possess the P-configuration (i.e., with the acetyl portions below the anthraquinone plane) at both axes in the case of joziknipholone A, whereas in joziknipholone B, the knipholone part was found to be M-configured. Joziknipholones A and B are active against the chloroquine resistant strain K1 of the malaria pathogen, Plasmodium falciparum, and show moderate activity against murine leukemic lymphoma L5178y cells.

Sediment phosphorus extractants for phosphorus-31 nuclear magnetic resonance analysis: a quantitative evaluation.

The influence of pre-extractant, extractant, and post-extractant on total extracted amounts of P and organic P compound groups measured with 31P nuclear magnetic resonance (31P-NMR) in lacustrine sediment was examined. The main extractants investigated were sodium hydroxide (NaOH) and sodium hydroxide ethylenediaminetetraacetic acid (NaOH-EDTA) with bicarbonate buffered dithionite (BD) or EDTA as pre-extractants. Post extractions were conducted using either NaOH or NaOH-EDTA, depending on the main extractant. Results showed that the most efficient combination of extractants for total P yield was NaOH with EDTA as pre-extractant, yielding almost 50% more than the second best procedure. The P compound groups varying the most between the different extraction procedures were polyphosphates and pyrophosphates. NaOH with BD as pre-extractant was the most efficient combination for these compound groups.

Aluminum control of phosphorus sorption by lake sediments.

Release of reactive (phosphate-like) phosphorus (P) from freshwater sediments represents a significant internal P source for many lakes. Hypolimnetic P release occurs under reducing conditions that cause reductive dissolution of ferric hydroxide [Fe(OH)3]. This hypolimnetic P release may be naturally low or artificially reduced by sediment with naturally high or artificially elevated concentrations of aluminum hydroxide [Al(OH)3]. We presentfield and laboratory data for a common extraction analysis of sediments from 43 lakes differing in trophic status, pH regime, climate, and P loading. The results indicate that a simple sequential extraction of sediment may be a useful predictor of sediment's ability to release P. Sequential extractions of sediment P, Al, and Fe by water (H2O), bicarbonate-dithionite (BD), and NaOH (at 25 degrees C) showed that negligible amounts of P would be released from lake sediments during hypolimnetic anoxia if either (1) the molar Al(NaOH-25):Fe(BD) ratio is > 3 or (2) the molar Al(NaOH-25):P(H2O+BD) ratio is > 25. These ratios can be used as operational targets for estimation of sediment P release potential and Al dosing of P-rich sediment to prevent hypolimnetic P release under anoxic conditions.

Cyanide binding at the non-heme Fe2+ of the iron-quinone complex of photosystem II: at high concentrations, cyanide converts the Fe2+ from high (S = 2) to low (S = 0) spin.

The primary electron acceptor complex of photosystem II, QAFe2+, can bind a number of small molecules at the iron site, including cyanide [Koulougliotis, D., Kostopoulos, T., Petrouleas, V., & Diner, B. A. (1993) Biochim. Biophys. Acta 1141, 275-282)]. In the presence of NaCN (30-300 mM) at pH 6.5, the reduced state, QA-Fe2+, produced either by illumination at < or = 200 K or by reduction in the dark with sodium dithionite, is characterized by a g = 1.98 EPR signal. The light- or dithionite-induced g = 1.98 signal decays with increasing pH above 6.5 and is almost totally absent at pH 8.1 and NaCN concentrations above 300 mM. However, at high pH (8.1), the g = 1.98 signal still forms transiently before it decays with a t1/2 of approximately 30 min in spinach BBY preparations treated with 100 mM NaCN. Complementary to the disappearance of the g = 1.98 signal with increasing pH or incubation time, a new EPR signal develops at g = 2.0045. This signal has the characteristics of the semiquinone, QA-, uncoupled from its magnetic interaction with the iron. Prolonged incubation of a high pH, high cyanide treated sample in a cyanide-free medium at pH 6 restores the ability of the sample to develop the cyanide-induced g = 1.98 signal at pH 6.5. This indicates that the iron is not physically dissociated during the high pH cyanide treatment. The high pH, high cyanide effects are accompanied by the conversion of the characteristic Fe2+ (S = 2) Mössbauer doublet [isomer shift (Fe) = 1.19 mm/s, quadrupole splitting = 2.95 mm/s] to a new one with parameters (isomer shift = 0.26 mm/s, quadrupole splitting = 0.36 mm/s) characteristic of an Fe2+(S = 0) state.(ABSTRACT TRUNCATED AT 250 WORDS)

The use of N-(7-nitrobenz-2-oxa-1,3-diazole-4-yl)-labeled lipids in determining transmembrane lipid distribution.

Transbilayer lipid distribution of small unilamellar vesicles (SUVs) and large unilamellar vesicles (LUVs) was measured using 31P-nuclear magnetic resonance (NMR) spectroscopy, chemical modification with 2,4,6-trinitrobenzene sulfonic acid (TNBS) and dithionite reduction of N-(7-nitrobenz-2-oxa-1,3-diazole-4-yl)-labeled lipid (NBD-lipid). The dithionite assay was the most reproducible of the three assays, with 1.2% error for SUVs and 3.9% error for LUVs. The dithionite assay also agreed best with theoretical inner:outer leaflet ratios, based on vesicle diameters determined by electron microscopy (Thomas et al. (1989) Biochem. Biophys. Acta 978, 85-90). Dithionite assay measurements were within 2.7% of theoretical ratios for SUVs and 2.3% for LUVs, while the NMR assay for SUVs was 14% lower than theoretical ratios and 23% lower for LUVs. The accuracy of NBD-lipids as markers for total transbilayer lipid was investigated. NBD-labeled phosphatidylserine, phosphatidylcholine and phosphatidylglycerol were accurate markers for total transbilayer lipid distribution, as their distributions were in close agreement with theoretical ratios. However, NBD-labeled phosphatidylethanolamine displayed a slight preference for the inner leaflet at low mole fractions of phosphatidylethanolamine, while native phosphatidylethanolamine showed a preference for the outer leaflet at the same concentration. NBD-labeled phosphatidic acid also showed a slight preference for the inner leaflet. We conclude that although dithionite-based assessment of NBD-labeled lipids across membrane bilayers can be a powerful analytical tool, caution must be used in the interpretation of results.

Inhibition of O2-. generating oxidase of neutrophils by iodonium biphenyl in a cell free system: effect of the redox state of the oxidase complex.

The conditions of inhibition of neutrophil O2-. generating oxidase by iodonium biphenyl (IBP) were studied. In a cell free system of oxidase activation consisting of neutrophil membranes and cytosol, GTP-gamma-S, Mg2+ and arachidonic acid, the inhibitory effect of IBP depended on the redox conditions of the medium. Inhibition was observed when the medium was supplemented with dithionite or NADPH. When the cell free system was incubated with IBP in the absence of reducing agents, full oxidase activity was recovered after removal of free IBP by gel filtration. Bovine neutrophil membranes, but not cytosol, contained component(s) sensitive to IBP. Upon treatment of neutrophil membranes by IBP followed by reduction, the spectrum of reduced cytochrome b558 was modified, suggesting that cytochrome b558 is a target site for IBP.

Colorimetry and constant-potential coulometry determinations of transferrin-bound iron, total iron-binding capacity, and total iron in serum containing iron-dextran, with use of sodium dithionite and alumina columns.

After the parenteral administration of iron-dextran (imferon), the increased total iron concentrations in serum can be determined by atomic absorption spectroscopy and by colorimetric methods involving sodium dithionite, which reductively dissociates iron from the dextran complex. We report that constant-potential coulometry detects only about 55-70% of dextran-bound iron before dithionite reduction and variable amounts after reaction with the reducing agent. In addition, we have developed a procedure for determining transferrin-bound iron, total iron-binding capacity (TIBC), total iron, and dextran-bound iron with the Kodak Ektachem colorimetric system. In determining total serum iron, the sample is first mixed with sodium dithionite, which rapidly dissociates all dextran-bound iron, but does not remove iron from either transferrin or hemoglobin. After the mixture is applied to an Ektachem slide, transferrin-bound iron is released at pH 4 and is detected together with the iron previously bound to dextran. TIBC is determined by mixing serum with ferric citrate in moderate excess and filtering through a small alumina (Al2O3) column, which binds excess free iron and iron-dextran; the iron in the column eluate represents the TIBC. Transferrin-bound iron is determined by applying diluted serum without added ferric citrate to an alumina column and measuring the iron in the column eluate. Dextran-bound iron is equivalent to the difference between total and transferrin-bound iron. Using this method, we found that transferrin iron-binding sites are saturated in vitro by excess iron-dextran less efficiently than by ferric citrate.

Murine cytotoxic activated macrophages inhibit aconitase in tumor cells. Inhibition involves the iron-sulfur prosthetic group and is reversible.

Previous studies show that cytotoxic activated macrophages cause inhibition of DNA synthesis, inhibition of mitochondrial respiration, and loss of intracellular iron from tumor cells. Here we examine aconitase, a citric acid cycle enzyme with a catalytically active iron-sulfur cluster, to determine if iron-sulfur clusters are targets for activated macrophage-induced iron removal. Results show that aconitase activity declines dramatically in target cells after 4 h of co-cultivation with activated macrophages. Aconitase inhibition occurs simultaneously with arrest of DNA synthesis, another early activated macrophage-induced metabolic change in target cells. Dithionite partially prevents activated macrophage induced aconitase inhibition. Furthermore, incubation of injured target cells in medium supplemented with ferrous ion plus a reducing agent causes near-complete reconstitution of aconitase activity. The results show that removal of a labile iron atom from the [4Fe-4S] cluster, by a cytotoxic activated macrophage-mediated mechanism, is causally related to aconitase inhibition.

Characterization of the selenium-substituted 2 [4Fe-4Se] ferredoxin from Clostridium pasteurianum.

The sulfur atoms of the two [4Fe-4S] clusters present in the ferredoxin from Clostridium pasteurianum have been replaced by selenium. The substitution is readily carried out by incubating the apoferredoxin with excess amounts of Fe3+, selenite, and dithiothreitol under anaerobic conditions. The UV-visible absorption spectrum of the Se-substituted ferredoxin, the core extrusion of its active sites, and analyses of its iron and selenium contents show that it contains two [4Fe-4Se] clusters. The Se-substituted ferredoxin is considerably less resistant to oxygen or to acidic and alkaline pH than the native ferredoxin: the half-lives of the former are 20-500 times shorter than those of the latter. The native ferredoxin and the Se-substituted ferredoxin display similar kinetic properties when used as electron donors to the hydrogenase from C. pasteurianum. It is of note, however, that the Km and Vmax values are lower for the 2[4Fe-4Se] ferredoxin than for the 2[4Fe-4S] ferredoxin. Reductive and oxidative titrations with dithionite and with thionine, respectively, show that both ferredoxins are two-electron carriers. The redox potentials of the ferredoxins have been measured by equilibrating them with the H2/H+ couple via hydrogenase: values of -423 and -417 mV have been found for the 2[4Fe-4S] ferredoxin and 2[4Fe-4Se] ferredoxin, respectively. Ferredoxins containing both chalcogenides in their [4Fe-4X] (X = S, Se) clusters have been prepared by reconstitution reactions involving mixtures of sulfide and selenide: the latter experiments show that sulfide and selenide are equally reactive in the incorporation of [4Fe-4X] (X = S, Se) sites into ferredoxin. The present report, together with former studies, establishes the general feasibility of the Se/S substitution in [2Fe-2S] and in [4Fe-4S] clusters of proteins and of synthetic analogues.

Nutritional studies with Pseudomonas aeruginosa grown on inorganic sulfur sources.

Pseudomonas aeruginosa was grown on a succinate-basal salts medium supplemented with various inorganic sulfur compounds as its sole source of sulfur. The organism was able to grow on the sodium salts of sulfide, thiosulfate, tetrathionate, dithionite, metabisulfite, sulfite, or sulfate, but not on those of dithionate. Analyses of the culture media after 24 h of growth indicated accumulation of sulfate from each inorganic sulfur source except sulfate. Manometric studies with resting cells obtained by growth on each of these sulfur sources yielded net oxygen uptake for all substrates except sulfite and dithionate. Similar results were obtained with extracts from these cells by spectrophotometric techniques. Thiosulfate oxidase activity appeared to be induced by growth on sulfide, thiosulfate, or tetrathionate, with little or no activity observed when cells were grown on inorganic sulfur sources of higher oxidative states. Metabisulfite oxidase appeared to be associated with growth on all inorganic sulfur compounds. Rhodanese activity appeared to be constitutively present, and its activity, observed only in soluble fraction, seemed independent of the growth medium employed. Thiosulfate and tetrathionate oxidase activities were studied in greater detail than some of the other sulfur oxidases, and both were found to be distributed between particulate and soluble fractions.

Oxidation-reduction behavior of the heme c and heme d moieties of Pseudomonas aeruginosa nitrite reductase and the formation of an oxygenated intermediate at heme d1.

Dithionite reduced the heme c moiety of Pseudomonas nitrite reductase almost instantaneously, whereas the spectral change of heme d proceeded in two steps, requiring at least 15 min for completion. The final spectrum coincided well with that obtained by anaerobic reduction with ascorbate, during which a quasi oxidation-reduction equilibrium was established between the two heme groups. The difference in apparent redox potential was calculated to be 24 mV, heme d being more negative. When the enzyme was supplemented with a reductant and molecular oxygen, an oxygenated intermediate appeared at the heme d moiety.