Cadmium uptake kinetics in intact soybean plants

The absorption characteristics of Cd²⁺ by 10- to 12-day-old soybean plants (Glycine max cv Williams) were investigated with respect to influence of Cd concentration on adsorption to root surfaces, root absorption, transport kinetics and interaction with the nutrient cations Cu²⁺, Fe²⁺, Mn²⁺, and Zn²⁺. The fraction of nonexchangeable Cd bound to roots remained relatively constant at 20 to 25% of the absorbed fraction at solution concentration of 0.0025 to 0.5 micromolar, and increased to 45% at solution concentration in excess of 0.5 micromolar. The exchangeable fraction represented 1.4 to 32% of the absorbed fraction, and was concentration dependent. Using dinitrophenol as a metabolic inhibitor, the `metabolically absorbed' fraction was shown to represent 75 to 80% of the absorbed fraction at concentration less than 0.5 micromolar, and decreased to 55% at 5 micromolar. At comparatively low Cd concentrations, 0.0025 to micromolar 0.3, root absorption exhibited two isotherms with K₂ values of 0.08 and 1.2 micromolar. Root absorption and transfer from root to shoot of Cd²⁺ was inhibited by Cu²⁺, Fe²⁺, Mn²⁺, and Zn²⁺. Analyses of kinetic interaction of these nutrient cations with Cd²⁺ indicated that Cu²⁺, Fe²⁺, Zn²⁺, and possibly Mn²⁺ inhibited Cd absorption competitively suggesting an involvement of a common transport site or process.     

Absorption of copper, zinc, and manganese by sugarcane leaf tissue

The absorption of Cu²⁺, Zn²⁺, and Mn²⁺ by leaf tissue of 4-month old sugarcane plants (Saccharum officinarum L., var. H53-263) has been investigated. After the “apparent free space” fraction was desorbed, the absorption of Cu²⁺, Mn²⁺, and Zn²⁺ yielded a curve typical of many ion uptake processes when measured as a function of the external concentration. However, only 1 absorption mechanism was evident for each cation. The pH optimum for Cu²⁺ and Zn²⁺ uptake was 5.0 to 6.0, whereas that for Mn²⁺ absorption was 4.5 to 6.0. Absorption was competitively inhibited by H⁺, and this inhibition was reversible when 0.5 mm Ca²⁺ was present. Cu²⁺ and Zn²⁺ were absorbed through the same carrier sites, as concluded from their mutually competitive activities. Mn²⁺ was absorbed through a second, independent mechanism. Uptake of each cation was strongly inhibited by uncouplers of oxidative phosphorylation, by Amytal and Nembutal², by 5 × 10⁻²m succinate, and by ADP and P_i. Absorption of Cu²⁺, Zn²⁺, and Mn²⁺ was concluded to be coupled to oxidative phosphorylation, and specifically to energy-conservation Site I.     

Use of ion chromatography for the determination of selected metals in blood serum of patients with type 2 diabetes

Ion chromatography followed by microwave-induced acid digestion was used to evaluate the serum levels of Fe³⁺, Cu²⁺, Ni²⁺, Zn²⁺, and Mn²⁺ in patients with diagnosed type 2 diabetes and in healthy controls. Recoveries ranged from 98.0% to 102% for Fe³⁺, from 89.9% to 100% for Cu²⁺, from 87.9% to 102% for Zn²⁺, and from 89.6% to 102% for Mn²⁺ were determined by examining samples spiked with various amounts of all the studied ions. The time of mineralization longer than 28 min did not affect the assay values. Precision was assessed at four unique concentrations in replicates of six, on four separate occasions. RSD was determined to be 1.16% for Fe³⁺, 5.20% for Cu²⁺, 2.8% for Zn²⁺, and 3.75% for Mn²⁺. The accuracy results (values of RSD) were as follows: 5.16% for Fe³⁺, 6.35% for Cu²⁺, 4.9% for Zn²⁺, and 7.23% for Mn²⁺.The statistical analysis confirmed that mean concentrations of Fe³⁺ and Zn²⁺ did not differ significantly from analogous values in the control group. Patients who additionally suffered from hypertension had higher copper concentrations compared with diabetic patients. For diabetics the presence of Mn²⁺ was not stated (LOD values amounting to 0.006 μg/mL). Ni²⁺ was not detectable for either the studied group or the control group (LOD=0.006 μg/mL).     

Probing the coordination properties of glutathione with transition metal ions (Cr2+,Mn2+,Fe2+,Co2+, Ni2+,Cu2+,Zn2+,Cd2+,Hg2+) by density functional theory

Complexes formed by reduced glutathione (GSH) with metal cations (Cr²⁺, Mn²⁺,Fe²⁺,Co²⁺,Ni²⁺,Cu²⁺,Zn²⁺,Cd²⁺,Hg²⁺) were systematically investigated by the density functional theory (DFT). The results showed that the interactions of the metal cations with GSH resulted in nine different stable complexes and many factors had an effect on the binding energy. Generally, for the same period of metal ions, the binding energies ranked in the order of Cu²⁺>Ni²⁺>Co²⁺>Fe²⁺>Cr²⁺>Zn²⁺>Mn²⁺; and for the same group of metal ions, the general trend of binding energies was Zn²⁺>Hg²⁺>Cd²⁺. Moreover, the amounts of charge transferred from S or N to transition metal cations are greater than that of O atoms. For Fe²⁺,Co²⁺,Ni²⁺,Cu²⁺,Zn²⁺,Cd²⁺ and Hg²⁺ complexes, the values of the Wiberg bond indices (WBIs) of M-S (M denotes metal cations) were larger than that of M-N and M-O; for Cr²⁺ complexes, most of the WBIs of M-O in complexes were higher than that of M-S and M-N. Furthermore, the changes in the electron configuration of the metal cations before and after chelate reaction revealed that Cu²⁺, Ni²⁺,Co²⁺ and Hg²⁺ had obvious tendencies to be reduced to Cu⁺,Ni⁺,Co⁺ and Hg⁺ during the coordination process.     

Inhibition of the biosynthesis ofN-acetylneuraminic acid by metal ions and seleniumin vitro

In liver homogenate the biosynthesis ofN-acetylneuraminic acid usingN-acetylglucosamine as precursor can be followed stepwise by applying different chromatographic procedures. In this cell-free system 16 metal ions (Zn²⁺, Mn²⁺, La³⁺, Co²⁺, Cu²⁺, Hg²⁺, VO ₃ ^– , Pb²⁺, Ce³⁺, Cd²⁺, Fe²⁺, Fe³⁺, Al³⁺, Sn²⁺, Cs⁺ and Li⁺) and the selenium compounds, selenium(IV) oxide and sodium selenite, have been checked with respect to their ability to influence a single or possible several steps of the biosynthesis ofN-acetylneuraminic acid. It could be shown that the following enzymes are sensitive to these metal ions (usually applied at a concentration of 1 mmoll^–1):N-acetylglucosamine kinase (inhibited by Zn²⁺ and vandate), UDP-N-acetylglucosamine-2-epimerase (inhibited by zn²⁺, Co²⁺, Cu²⁺, Hg²⁺, VO ₃ ^– , Pb²⁺, Cd²⁺, Fe³⁺, Cs⁺, Li⁺, selenium(IV) oxide and selenite), andN-acetylmannosamine kinase (inhibited by Zn²⁺, Cu²⁺, Cd²⁺, and Co²⁺). Dose dependent measurements have shown that Zn²⁺, Cu²⁺ and selenite are more efficient inhibitors of UDP-N-acetylglucosamine-2-epimerase than vanadate. As for theN-acetylmannosamine kinase inhibition, a decreasing inhibitory effect exists in the following order Zn²⁺, Cd²⁺, Co²⁺ and Cu²⁺. In contrast, La³⁺, Al³⁺ and Mn²⁺ (1 mmoll^–1) did not interfere with the biosynthesis ofN-acetylneuraminic acid. Thus, the conclusion that the inhibitory effect of the metal ions investigated cannot be regarded as simply unspecific is justified.Dedicated to Professor Theodor Günther on the occasion of his 60th birthday     

Purification and biochemical characterization of two extracellular peroxidases from Phanerochaete chrysosporium responsible for lignin biodegradation

Two extracellular peroxidases from Phanerochaete chrysosporium, namely a lignin peroxidase (LiP) and manganese peroxidase (MnP), were purified simultaneously by applying successively, ultrafiltration, ion-exchange and gel filtration chromatography. LiP and MnP have a molecular mass of 36 and 45 kDa, respectively. The optimal pHs for LiP and MnP activities were 3.0 and 4.5, respectively. Both peroxidases showed maximal activity at 30 °C and moderate thermostability. MnP activity was strongly inhibited by Fe²⁺, Zn²⁺, Mg²⁺ and Hg²⁺, and enhanced by Mn²⁺, Ca²⁺ and Cu²⁺. LiP activity was enhanced by Ca²⁺, Na⁺ and Co²⁺ and it was inhibited in the presence of K⁺, Hg⁺, Fe²⁺, Mg²⁺ and high concentrations of Cu²⁺ and Zn²⁺. The K_m and V_max for LiP toward veratryl alcohol as a substrate were 0.10 mM and 15.2 U mg⁻¹, respectively and for MnP toward Mn²⁺, they were respectively 0.03 mM and 25.5 U mg⁻¹. The two peroxidases were also able to break down rice lignin in a small-scale solid state treatment system. Data suggest these two peroxidases may be considered as potential candidates for the development of enzyme-based technologies for lignin degradation.     

Iron transport by proteoliposomes containing mitochondrial F1Fo ATP synthase isolated from rat heart

In this work, we present evidence of Fe²⁺ transport by rat heart mitochondrial F₁F_o ATP synthase. Iron uptake by the vesicles containing the enzyme was concentration- and temperature-dependent, with an optimum temperature of 37 °C. Both ATP and ADP stimulated iron uptake in a concentration-dependent manner, whereas AMP, AMPPCP, and mADP did not. Inhibitors of the enzyme, oligomycin, and resveratrol similarly blocked iron transport. The iron uptake was confirmed by inhibition using specific antibodies against the α, β, and c subunits of the enzyme. Interestingly, slight transport of common divalent and trivalent metal ions such as Mg⁺², Ca⁺², Mn⁺², Zn⁺², Cu⁺², Fe⁺³, and Al⁺³ was observed. Moreover, Cu⁺², even in the nM range, inhibited iron uptake and attained maximum inhibition of approximately 56%. Inorganic phosphate (Pi) in the medium exerted an opposite effect depending on the type of adenosine nucleotide, which was suppressed with ATP, but enhanced with ADP. A similarly stimulating effect of ATP and ADP with an inverse effect of Pi suggests that the activity of ATPase and ATP synthase may be associated with iron uptake in a different manner, probably via antiport of H⁺.     

Protective effects of pretreatment with Fe2+, Cu2+, and Rb+ on phoxim poisoning in silkworm,Bombyx mori

BackgroundPhoxim is a widely used organophosphorus pesticide in agriculture. People are paying more and more attention to its toxicity. At present, there is no appropriate way to solve the phoxim poisoning of silkworm, which severely affected the development of sericulture. Fe²⁺, Cu²⁺, Rb⁺ exerted their biological effects through various forms in vivo.MethodsTo evaluate the effect of Fe²⁺/Cu²⁺/Rb⁺ on phoxim poisoning in silkworm, Bombyx mori were treated with fresh mulberry leaves soaked in 2.5 mg/L phoxim for 2 min with 50 mg/L FeCl₂, 150 mg/L CuCl₂, or 0.5 mg/L RbCl from 5 days of the fifth-instar silkworm.ResultsFe²⁺, Cu²⁺, and Rb⁺ pretreatments significantly inhibited the phoxim-induced reduction of survival rate and alleviated the phoxim-induced poisoning symptoms. The protective effects of Fe²⁺, Cu²⁺, and Rb⁺ on phoxim poisoning might be due to their enhancement of superoxide dismutase (SOD), catalase (CAT), and carboxylesterase (CarE) in the hemolymph and fat body of silkworm. This enhancement might reduce reactive oxygen species (ROS) accumulation and oxidative stress (OS) caused by phoxim poisoning. Thereby it reduced the damage to silkworm tissues and cells.ConclusionsThese results showed that Fe²⁺, Cu²⁺, and Rb⁺ treatments protected the silkworm from phoxim poisoning by directly enhancing the activity of SOD, CAT, and CarE enzymes and reducing oxidative stress, but not dependent on the high expression of CYP genes. The use of Fe²⁺, Cu²⁺, and Rb⁺ to enhance the activity of SOD, CAT, and CarE enzymes may be an underlying effective way to solve phoxim poisoning in the silkworm industry.     

Effect of divalent metal ions on the microsomal aniline hydroxylase system of rainbow trout

The ability of trout to metabolize aniline in vitro in the presence of some divalent metal ions was investigated in the liver microsomes of rainbow trout, Salmo gairdneri. Trout liver microsomes were highly capable of catalyzing aniline hydroxylation to p-aminophenol with a specific activity of 0.068 nmoles/min per mg of microsomal protein in potassium phosphate buffer, pH 7.4 at 25°C. The activity of the aniline hydroxylase system was competitively inhibited by Hg⁺², Ni⁺², Cd⁺², and Zn⁺², while Cu⁺² and Fe⁺³ seemed to inhibit the activity noncompetitively at 1 mM aniline concentrations. IC₅₀ values at fixed aniline concentration were estimated to be 0.45 mM for Hg⁺², Ni⁺², and Cd⁺², 1.8 mM for Zn⁺² and Fe⁺³, and 1.3 mM for Cu⁺². Eadie-Hofstee plots gave identical V_max values of approximately 0.046 nmol/min per mg of protein while K_m values were increased in the presence of Hg⁺², Ni⁺², CD⁺², and Zn⁺², indicating competitive inhibition. Both K_m and V_max values were affected by Fe⁺³ and Cu⁺², suggesting noncompetitive inhibition. K_i values extracted from the Dixon plots were determined t be 0.23, 0.43, and 0.65 mM for Hg⁺², Ni⁺², and Cd⁺², respectively, providing the most effective inhibition on the aniline hydroxylase system among studied metal ions. The K_i values were much higher in the presence of others. The results indicate a selective inhibition of the aniline hydroxylase system of trout liver microsomes by divalent metal ions. © 1997 John Wiley & Sons, Inc.     

Substrate Profile and Metal-ion Selectivity of Human Divalent Metal-ion Transporter-1

Divalent metal-ion transporter-1 (DMT1) is a H⁺-coupled metal-ion transporter that plays essential roles in iron homeostasis. DMT1 exhibits reactivity (based on evoked currents) with a broad range of metal ions; however, direct measurement of transport is lacking for many of its potential substrates. We performed a comprehensive substrate-profile analysis for human DMT1 expressed in RNA-injected Xenopus oocytes by using radiotracer assays and the continuous measurement of transport by fluorescence with the metal-sensitive PhenGreen SK fluorophore. We provide validation for the use of PhenGreen SK fluorescence quenching as a reporter of cellular metal-ion uptake. We determined metal-ion selectivity under fixed conditions using the voltage clamp. Radiotracer and continuous measurement of transport by fluorescence assays revealed that DMT1 mediates the transport of several metal ions that were ranked in selectivity by using the ratio I_max/K_0.5 (determined from evoked currents at −70 mV): Cd²⁺ > Fe²⁺ > Co²⁺, Mn²⁺ ≫ Zn²⁺, Ni²⁺, VO²⁺. DMT1 expression did not stimulate the transport of Cr²⁺, Cr³⁺, Cu⁺, Cu²⁺, Fe³⁺, Ga³⁺, Hg²⁺, or VO⁺. ⁵⁵Fe²⁺ transport was competitively inhibited by Co²⁺ and Mn²⁺. Zn²⁺ only weakly inhibited ⁵⁵Fe²⁺ transport. Our data reveal that DMT1 selects Fe²⁺ over its other physiological substrates and provides a basis for predicting the contribution of DMT1 to intestinal, nasal, and pulmonary absorption of metal ions and their cellular uptake in other tissues. Whereas DMT1 is a likely route of entry for the toxic heavy metal cadmium, and may serve the metabolism of cobalt, manganese, and vanadium, we predict that DMT1 should contribute little if at all to the absorption or uptake of zinc. The conclusion in previous reports that copper is a substrate of DMT1 is not supported.     

Inhibition of Na+/K+-ATPase and Mg2+-ATPase by metal ions and prevention and recovery of inhibited activities by chelators

Kinetics and inhibition of Na⁺/K⁺-ATPase and Mg²⁺-ATPase activity from rat synaptic plasma membrane (SPM), by separate and simultaneous exposure to transition (Cu²⁺, Zn²⁺, Fe²⁺ and.Co²⁺) and heavy metals (Hg²⁺and Pb²⁺) ions were studied. All investigated metals produced a larger maximum inhibition of Na⁺/K⁺-ATPase than Mg²⁺-ATPase activity. The free concentrations of the key species (inhibitor, MgATP^{2 ?} , MeATP^{2 ?} ) in the medium assay were calculated and discussed. Simultaneous exposure to the combinations Cu²⁺/Fe²⁺ or Hg²⁺/Pb²⁺caused additive inhibition, while Cu²⁺/Zn²⁺ or Fe²⁺/Zn²⁺ inhibited Na⁺/K⁺-ATPase activity synergistically (i.e., greater than the sum metal-induced inhibition assayed separately). Simultaneous exposure to Cu²⁺/Fe²⁺ or Cu²⁺/Zn²⁺ inhibited Mg²⁺-ATPase activity synergistically, while Hg²⁺/Pb²⁺ or Fe²⁺/Zn²⁺ induced antagonistic inhibition of this enzyme. Kinetic analysis showed that all investigated metals inhibited Na⁺/K⁺-ATPase activity by reducing the maximum velocities (V_max) rather than the apparent affinity (K_m) for substrate MgATP^2-, implying the noncompetitive nature of the inhibition. The incomplete inhibition of Mg²⁺-ATPase activity by Zn²⁺, Fe²⁺ and Co²⁺ as well as kinetic analysis indicated two distinct Mg²⁺-ATPase subtypes activated in the presence of low and high MgATP^{2 ?} concentration. EDTA, L-cysteine and gluthathione (GSH) prevented metal ion-induced inhibition of Na⁺/K⁺-ATPase with various potencies. Furthermore, these ligands also reversed Na⁺/K⁺-ATPase activity inhibited by transition metals in a concentration-dependent manner, but a recovery effect by any ligand on Hg²⁺-induced inhibition was not obtained.     

Effect of Fe2+, Mn2+, Zn2+ and Pb2+ on H+/K+ fluxes in excisedPistia stratiotes roots

Pistia stratiotes is used for the epuration of domestic sewage in the Biyem Assi phytopurification station. During the process, Fe²⁺, Mn²⁺, Zn²⁺ and Pb²⁺ are absorbed in substantial amounts by the plant. These metals modify the H⁺/K⁺ exchange system at the root level. H⁺ efflux is inhibited by Fe²⁺ and by Zn²⁺ and enhanced by Mn²⁺ and Pb²⁺. K⁺ influx is inhibited by Fe²⁺, by Zn²⁺ and by Pb²⁺ and enhanced by Mn²⁺. It is shown that the purification capacity ofPistia stratiotes can vary with the composition of the heavy metals in the surrounding medium.     

A new heterobinuclear FeIIICuII complex with a single terminal FeIII–O(phenolate) bond. Relevance to purple acid phosphatases and nucleases

A novel heterobinuclear mixed valence complex [Fe^IIICu^II(BPBPMP)(OAc)₂]ClO₄, 1, with the unsymmetrical N₅O₂ donor ligand 2-bis[{(2-pyridylmethyl)aminomethyl}-6-{(2-hydroxybenzyl)(2-pyridylmethyl)}aminomethyl]-4-methylphenol (H₂BPBPMP) has been synthesized and characterized. A combination of data from mass spectrometry, potentiometric titrations, X-ray absorption and electron paramagnetic resonance spectroscopy, as well as kinetics measurements indicates that in ethanol/water solutions an [Fe^III–()OH–Cu^IIOH₂]⁺ species is generated which is the likely catalyst for 2,4-bis(dinitrophenyl)phosphate and DNA hydrolysis. Insofar as the data are consistent with the presence of an Fe^III-bound hydroxide acting as a nucleophile during catalysis, 1 presents a suitable mimic for the hydrolytic enzyme purple acid phosphatase. Notably, 1 is significantly more reactive than its isostructural homologues with different metal composition (Fe^IIIM^II, where M^II is Zn^II, Mn^II, Ni^II, or Fe^II). Of particular interest is the observation that cleavage of double-stranded plasmid DNA occurs even at very low concentrations of 1 (2.5 M), under physiological conditions (optimum pH of 7.0), with a rate enhancement of 2.7×10⁷ over the uncatalyzed reaction. Thus, 1 is one of the most effective model complexes to date, mimicking the function of nucleases.Electronic Supplementary Material Supplementary material is available for this article at .     

Effect of Sesbania,Azolla and rice straw incorporation on the kinetics of NH4, K,Fe, Mn,Zn and P in some flooded rice soils

In a greenhouse study, with and without rice plants, of five flooded Philippine rice soils whose organic C (OC) content varied from 0.5 to 3.6%, incorporation ofSesbania rostrata, Azolla microphylla and rice straw affected the kinetics of soil solution NH ₄ ⁺ −N, K⁺, Fe²⁺, Mn²⁺, Zn²⁺, and P. Sesbania and Azolla increased NH ₄ ⁺ −N concentration above the control treatment, whereas rice straw depressed it. In all soils Azolla released less NH ₄ ⁺ −N than Sesbania. The apparent net N release depended on the soil and ranged from 44–81% for Sesbania and 27–52% for Azolla. These effects persisted throughout the growth of IR36. Soil solution and exchangeable NH ₄ ⁺ −N increased initially but levelled off between 30 to 80 days and between 20 to 40 days after flooding (DF), respectively. With rice, soil solution NH ₄ ⁺ −N concentration, reached a peak at 15–40 DF and declined to very low levels (<4mg L⁻¹). In the 3 soils of low OC content nitrogen derived from green manure ranged from 34–53% and the apparent revovery of added green manure N varied from 29–67%. Almost all N released from both Azolla and Sesbania were recovered in the rice plant in all soils except Concepcion with only 77%. The concentration of K⁺, Fe²⁺, Mn²⁺ and P in the soil solution were higher with rice straw than Sesbania and Azolla in all soils except Hanggan which showed no change in Fe²⁺ and Mn²⁺ but increased K⁺ and P. In general, rice straw, Sesbania and Azolla decreased Zn²⁺ concentration in all soils.     

Influence of some trace metals and stimulants on citric acid production from brewery wastes by Aspergillus niger

The addition of increasing levels of Mn²⁺, Fe³⁺, Zn²⁺, Co²⁺, Cu²⁺, Ca²⁺, sodium monofluoracetate and methanol during citric acid surface fermentation of spent grain liquor by Aspergillus niger (ATCC 9142) was investigated. For spent grain liquor the addition of 51 ppb Mn²⁺, 5 ppb Fe³⁺, 75 ppb Zn²⁺ and 4% (v/va) methanol caused a 4.9, 1.9, 10.9 and 16.8% increase in citric acid yield respectively. In all other fermentations the yield of citric acid was decreased whereas the biomass production in some cases was increased.     

Levels and sub-cellular distribution of physiologically important metal ions in neuronal cells cultured from chick embryo cerebral cortex

Mg²⁺, Ca²⁺, Mn²⁺, Zn²⁺, and Cu content of neurons from chick embryo cortex cultivated in chemically defined serum free growth medium was determined by energy dispersive X-ray fluorescence and atomic absorption spectroscopy. The intracellular volume of cultured neurons was determined to be 2.73 l/mg. Intracellular Mn²⁺, Fe²⁺, Zn²⁺, and Cu²⁺ in the cultivated neurons were 100–200 times the concentrations in the growth medium: Mg²⁺ and Ca²⁺ were 0.9 and 1.7 mM respectively, around 20 fold higher than in growth medium. Mg²⁺, Fe²⁺, Cu²⁺ and Zn²⁺ concentrations in neurons were in the range of ca. 300–600 M, approximately 2–3 times the values previously reported in glial cells; Ca²⁺ and Mn²⁺ content of the neurons were higher by 5 and 10 fold respectively compared to glial cells. In neurons, the subcellular distribution of Fe²⁺, Cu²⁺, and Mn²⁺ follows the rank order: cytosol>microsomes>mitochondria; for Zn²⁺ the distribution differs as following: cytosol >mitochondria>microsomes. Determination of the superoxide dismutase activities in the cultivated neurons indicated that the Mn²⁺ linked activity predominates whereas, the Cu-Zn dependent enzyme is dominant in glial cells. Enrichment of the culture medium with Mn²⁺ to 2.5 M enhanced the Mn-SOD by approximately 33% but Cu²⁺–Zn²⁺ enzyme activity was not modified. The high Mn²⁺ content, the capacity to accumulate Mn²⁺, and the predominancy of the Mn–SOD form observed in neurons is in accord with a fundamental functional role for this metal ion in this type of brain cells.     

Chemical speciation in wound fluid leading to enhanced bioavailability for healing

ABSTRACT

ECCLES computer modelling is used to speciate Ca²⁺, Zn²⁺, Cu²⁺, Mn²⁺ and Fe³⁺ in wound fluid samples analysed for total metal content using Potentiometric stripping analysis. Bioavailability of metals, related to lipid soluble net-neutral complexes, was assessed and found to be broadly similar to the speciation in blood plasma. These appear to be benefits worthy of clinical investigation of keeping the wound fluid at pH = 6.4 and of raising the total cysteine concentration levels in wound fluid.     

Nickel in plants: I. Uptake kinetics using intact soybean seedlings

The absorption of Ni²⁺ by 21-day-old soybean plants (Glycine max cv. Williams) was investigated with respect to its concentration dependence, transport kinetics, and interactions with various nutrient cations. Nickel absorption, measured as a function of concentration (0.02 to 100 μm), demonstrated the presence of multiple absorption isotherms. Each of the three isotherms conforms to Michaelis-Menten kinetics; kinetic constants are reported for uptake by the intact plant and for transfer from root to shoot tissues. The absorption of Ni²⁺ by the intact plant and its transfer from root to shoot were inhibited by the presence of Cu²⁺, Zn²⁺, Fe²⁺, and Co²⁺. Competition kinetic studies showed Cu²⁺ and Zn²⁺ to inhibit Ni²⁺ absorption competitively, suggesting that Ni²⁺, Cu²⁺, and Zn²⁺ are absorbed using the same carrier site. Calculated K_m and K_i constants for Ni²⁺ in the presence and absence of Cu²⁺ were 6.1 and 9.2 μm, respectively, whereas K_m and K_i constants were calculated to be 6.7 and 24.4 μm, respectively, for Ni²⁺ in the presence and absence of Zn²⁺. The mechanism of inhibition of Ni²⁺ in the presence of Fe²⁺ and Co²⁺ was not resolved by classical kinetic relationships.     

Purification and Characterization of Killer Toxin from a Marine Yeast <Emphasis Type="Italic">Pichia anomala</Emphasis> YF07b Against the Pathogenic Yeast in Crab

The molecular mass of the purified killer toxin from the marine killer yeast YF07b was estimated to be 47.0 kDa. The optimal pH and temperature of the purified killer toxin were 4.5 and 40°C, respectively. The toxin was activated by Ca²⁺, K⁺, Na⁺, Mg²⁺, Na⁺, and Co²⁺. However, Fe²⁺, Fe³⁺, Hg²⁺, Cu²⁺, Mn²⁺, Zn²⁺, and Ag⁺ acted as inhibitors in decreasing activity of the toxin. The toxin was strongly inhibited by phenylmethanesulphonyl fluoride (PMSF), iodoacetic acid, ethylenediaminetetraacetic acid, and 1,10-phenanthroline. The Km of the toxin for laminarin was 1.17 g L⁻¹. The toxin also actively hydrolyzed laminarin and killed the whole cells of the pathogenic yeast in crab.     

Super-high-affinity binding site for [3H]diazepam in the presence of Co2+, Ni2+, Cu2+, OR Zn2+

Chloride salts of Li⁺, Na⁺, K⁺, Mg²⁺, Ca²⁺, Cr³⁺, Mn²⁺, Fe²⁺, and Fe³⁺ had no effect on [³H]diazepam binding. Chloride salts of Co²⁺, Ni²⁺, Cu²⁺, and Zn²⁺ increased [³H]diazepam binding by 34 to 68% in a concentration-dependent fashion. Since these divalent cations potentiated the GABA-enhanced [³H]diazepam binding and the effect of each divalent cation was nearly additive with GABA, these cations probably act at a site different from the GABA recognition site in the benzodiazepine-receptor complex. Scatchard plots of [³H]diazepam binding without an effective divalent cation showed a single class of binding, with a Kd value of 5.3 mM. In the presence of 1 mM Co²⁺, Ni²⁺, Cu²⁺, or Zn²⁺, two distinct binding sites were evident with apparent Kd values of 1.0 nM and 5.7 nM. The higher-affinity binding was not detected in the absence of an effective divalent cation and is probably a novel, super-high-affinity binding site.