Nature of nickel complexes in Psychotria douarrei and other nickel-accumulating plants

The nature of nickel complexes in various Ni accumulating plant species, mainly from New Caledonia, was investigated by techniques including gel chromatography, ion-exchange chromatography, high performance liquid chromatography, and a combination of gas-liquid chromatography and mass spectrometry. Psychotria douarrei contained Ni complexed mainly (63%) as a negatively-charged malate complex balanced by a cationic aquo complex. Phyllanthus serpentinus had anionic Ni bound as 42% citrate and 4O% malate. All other species studied, contained Ni as an anionic citrate complex.     

The relationship between Candida species charge density and chitosan activity evaluated by ion-exchange chromatography

Chitosan, a natural biopolymer presents antifungal activity that seems to be dependent on the interaction of its cationic amino groups and yeast cell surface. In this work we used ion-exchange chromatography to assess the surface charge density of Candida species and subsequently to relate this with their sensitivity profile to chitosan. The ability of several strains from distinct Candida species to interact with strong anionic and cationic exchangers was tested and the yeasts charge surface was assessed by measuring the zeta potential. Our results showed that all the yeast cells tested presented no interaction with the cationic resin and a species-related pattern of interaction was observed with the anionic resin. Specifically, regarding the Q-Sepharose support, Candida glabrata showed the lower retention affinity, followed by Candida albicans, presenting Candida tropicalis an intermediate profile; Candida parapsilosis and Candida guilliermondii revealed a stronger ionic interaction. The yeasts retention synergy in the anionic resin corroborates with the zeta potential outcomes. The behavior observed fit with sensitivity patterns to chitosan as the most susceptible species to chitosan presented higher affinity to the anionic resin in contrast to the less sensitive ones (C. albicans and C. glabrata). This data confirms and reinforces that chitosan activity is probably mediated by an ionic reaction between its amino free groups and ionic charges at the cell surface.     

Speciation analysis of nickel in soil solutions and availability to oat plants

Soil solutions were collected for speciation analysis of nickel from a pot experiment with oats. Oat plants (Avena sativa L.) were grown on 3 soils differing in total amount and origin of nickel (Ni) (Luvisol, LS with 28 mg kg^-1; sludge amended Luvisol, LS+SS with 32 mg kg^-1; Cambisol, CS with 95 mg kg^-1). Results were compared with those for soil solutions obtained from corresponding unplanted pots. Separation methods were used for characterization of size and charge distribution and stability of the Ni species. In addition, short-term experiments were performed on the uptake rates of Ni by oat plants from the different soil solutions as well as from nutrient solutions with increasing concentrations of a synthetic chelator.The Ni concentrations in the soil solutions of unplanted soils increased in the order: LS5000 g mol^-1) was the predominant form, whereas in the other soils the low-molecular-size cationic and chelated Ni species (500–1000 g mol^-1) dominated in the soil solution. In the short-term uptake studies, the uptake rates of Ni from the solutions decreased in the order: nutrient solution > soil solutions, and in the latter in the order: LS>LS+SS>CS, which was inversely related to the concentrations of dissolved organic carbon in the soil solutions.The results demonstrate that Ni availability to plants is not only affected by total concentration of Ni in the soil solution and the rate of replenishment from the solid phase, but also by Ni species, which can differ considerably between soil types.     

The influence of ionic composition on the distribution of nematode species in springs of the province of Granada (Spain)

The ionic composition of 38 mineral springs in the province of Granada (Spain), and the distribution of 45 species of nematodes belonging to orders Monhysterida, Araeolaimida, Chromadorida and Enoplida were examined. Water chemistry is used to make two diagrams representing anionic and cationic composition. Diagrams for anionic composition (given the greater variance seen in the springs considered) are used to illustrate the distribution of individual species. The results obtained from species distribution and the correlation between species made it possible to group species which could be associated with springs where each of the anions considered predominated. A greater number of species groups was found to inhabit springs in which chloride concentrations was less than 50% of the total concentration of anions.     

Pancreatic anionic trypsin: evidence for the existence of a 30 kDa form.

1. An anionic form of trypsin has been isolated from pancreas of various species (rat, pig, dog and cow). 2. The purification procedure included affinity chromatography on STI-Sepharose 4B and ion-exchange chromatography on DEAE-Sephadex A-50. 3. The preparation was homogenous as checked by SDS-polyacrylamide slab gel electrophoresis, resulting in an estimated molecular weight of 30 kilodaltons (kDa) for this anionic form. 4. Antibodies against the anionic form from rat pancreas cross-reacted towards the anionic enzyme from porcine pancreas but not with the dog or bovine enzyme, nor with all the studied cationic forms. 5. Limited proteolysis of tubulin, a cytoskeletal protein, with an anionic or cationic form of trypsin showed striking differences in the size of produced peptides.     

The Distribution of Nickel, Copper, Zinc, and Iron in Tree Leaves

The distributions of nickel, copper, zinc, and iron in freshleaves from four tree species were studied using differentialoentrifugation of cell homogenates to fractionate the cell organellesand atomic absorption spectrophotometry to measure the concentrationsin the various fractions. Results from each species were closelysimilar. Approximately 90 per cent of the nickel was presentin the supernatant fraction while approximately 80 per centof the iron was present in the chloroplast fraction. Both copperand zinc had subcellular distributions similar to one anotherand contained appreciable amounts of the elements in both chloroplastand supernatant fractions. High-voltage paper electrophoretic separations of the solubleextracts of leaves showed that while copper, zinc, and ironexisted principally as anionic complexes, nickel existed asa cationio species. It was considered that this cationic formof nickel was unlikely to be an unchelated aquated cation.     

Isolation and identification of a citrato-complex of nickel from nickel-accumulating plants

An organometallic complex of nickel has been isolated from nickel-accumulating plants of New Caledonia and has been identified by a combination of infrared spectroscopy, high-voltage paper electrophoresis, gas-liquid chromatography and mass spectrometry. Most of the nickel in Sebertia acuminata, Homalium francii, Homalium guillainii, Homalium kanaliense, Hybanthus austrocaledonicus and Hybanthus caledonicus is found as a negatively-charged citratonickelate (II) complex with Ni(H₂O)₆²⁺ as the major cationic constituent.     

The relation between nickel and citric acid in some nickel-accumulating plants

A strong correlation has been found between the levels of nickel and citric acid in the leaves of 17 New Caledonian plant species which show a range of nickel-accumulating ability: Sebertia acuminata, Psychotria douarrei, Geissois pruinosa, Hybanthus austrocaledonicus, Hybanthus caledonicus, and 12 species of Homalium. Purified extracts of these plants contain nickel as a citrate complex. Three nickel-accumulating plants from beyond New Caledonia (Alyssum bertolonii, Alyssum serpyllifolium s.sp. lusitanicum and Pearsonia metallifera) do not contain unusually high levels of citrate, nor is citrate present in purified nickel-containing extracts of these species.     

Recognition of left-handed Z-DNA of short unmodified oligonucleotides under physiological ionic strength conditions

The left-handed Z-DNA form of the short unmodified alternating guanine-cytosine oligonucleotides, 5′-(dGdC)₂₄ and 5′-(dGdC)₁₈, was selectively detected under physiological ionic strength and pH conditions using the anionic nickel(II) porphyrin, NiTPPS. No spectroscopic signal was observed for NiTPPS with any right-handed oligonucleotides under identical conditions. The 48mer 5′-(dGdC)₂₄ Z-form was detected at concentrations as low as 100 nM. The binding of NiTPPS to the B- and Z-oligonucleotides was studied quantitatively by UV-vis absorption and circular dichroism spectroscopies. NiTPPS was found to be a universal DNA binder, with binding affinity and geometry depending on the ionic composition of the solution, rather than on the DNA helical twist. This is the first example of a successful spectroscopic detection of the Z-DNA of short unmodified oligonucleotides under physiological pH and ionic strength conditions.     

Structure-activity relationships of RGD mimetics as fibrinogen-receptor antagonists

The activities of a series of RGD mimetics, which contained a variety of cationic structures, for the inhibition of platelet aggregation and fibrinogen-receptor binding were measured. The stability of the coulombic ion-pairing complex of the model compounds with the acetate anion as a model for the receptor was calculated in terms of the ionic interaction energy. The results suggest that stability is one of the significant factors which govern the inhibitory potency of fibrinogen-receptor binding. The distance between cationic and anionic groups might also affect the potency. A compound which contained an amidinophenyl structure as the cationic moiety showed exceptionally high inhibitory activity, suggesting that some other factors, in addition to coulombic interaction and the distance, affect the potency.     

Antibacterial activity and biocompatibility of a chitosan-γ-poly(glutamic acid) polyelectrolyte complex hydrogel

In this study, we prepared a polyelectrolyte complex (PEC) hydrogel comprising chitosan as the cationic polyelectrolyte and γ-poly(glutamic acid) (γ-PGA) as the anionic polyelectrolyte. Fourier transform infrared spectroscopy revealed that ionic complex interactions existed in the chitosan-γ-PGA PEC hydrogels. The compressive modulus increased upon increasing the degree of complex formation in the chitosan-γ-PGA PEC hydrogel; the water uptake decreased upon increasing the degree of complex formation. At the same degree of complex formation, the compressive modulus was larger for the chitosan-dominated PEC hydrogels; the water uptake was larger for the γ-PGA-dominated ones. Scanning electron microscopy images revealed the existence of interconnected porous structures (pore size: 30-100 μm) in all of the chitosan-γ-PGA PEC hydrogels. The chitosan-γ-PGA PEC hydrogels also exhibited antibacterial activity against Escherichia coli and Staphylococcus aureus. In addition, in vitro cell culturing of 3T3 fibroblasts revealed that all the chitosan-γ-PGA PEC hydrogels were effective in promoting cell proliferation, especially the positively charged ones (chitosan-dominated). Therefore, the chitosan-γ-PGA polyelectrolyte hydrogel appears to have potential as a new material for biomedical applications.     

The charge distribution in the rough endoplasmic reticulum/golgi complex transitional area investigated by microinjection of charged tracers

The distribution of microinjected ferritin, ranging in charge from anionic to highly cationic, has been used to indicate differences in surface charge on the rough endoplasmic reticulum and the Golgi complex of intact cells. Highly cationic ferritins (HCF)(HCF1, pI 7.9-9.1; HCF2, pI 8.5-9.4; and HCF3.pI 9.5-10.1) were mostly bound and caused swelling of the rough endoplasmic reticulum. Cationic ferritin (CF) (pI 7.0-8.0) and anionic ferritin (AF) (pI 4.0-4.4) caused no changes in morphology. The distribution of these ferritins in the cytoplasmic space varied with their charge. Significantly more CF was bound to surfaces than was found in the free cytoplasmic space. Conversely, there was significantly more AF in the free cytoplasmic space than close to surfaces. Therefore, the intracellular surfaces are negatively charged. Comparison of the structures in the secretory pathway showed no differences in ferritin binding to transition vesicles, rough endoplasmic reticulum, Golgi saccules or secretory vesicles. The Golgi complex beads are not distinguished by their charge. It is therefore unlikely that charge differences play a role in regulating membrane-membrane interactions in this region of the secretory pathway.     

Peroxidases from alfalfa roots: Catalytic properties and participation in degradation of polycyclic aromatic hydrocarbons

From the roots and root exudates of 3-week-old plants of alfalfa (Medicago sativa L.), anionic and cationic peroxidases differing in principal physicochemical and catalytic properties were isolated and purified. Main features of anionic peroxidases detected in the roots and root exudates were identical. Phenanthrene present in the soil used for alfalfa growing influenced the number of forms and activity of peroxidases in crude enzyme preparations but did not affect the properties of pure enzymes. In the presence of a synthetic mediator, purified peroxidases can oxidize phenanthrene and its derivatives, including potential microbial metabolites of polycyclic aromatic hydrocarbons (PAH). The fact that the enzymes excreted in root exudates in a purified form can oxidase PAH proves their participation in degradation of PAH and their microbial metabolites in alfalfa rhizosphere. These new data indicate that the processes of plant and microbial degradation of pollutants in the rhizosphere are coupled; they are relevant to understanding the molecular mechanisms of degradation of persistent pollutants by plant-microbial complexes.     

Membrane fusion induced by mutual interaction of the two charge-reversed amphiphilic peptides at neutral pH

An anionic amphiphilic peptide and the charge-reversed cationic peptide are synthesized. They contain 20 amino acids with the same sequence except for 5 Glu residues for the anionic versus 5 Lys residues for the cationic peptides. Fusion of egg phosphatidylcholine large unilamellar vesicles is assayed with the fluorescent probes by the lipid mixing and the internal content mixing at neutral pH. The peptide mixture causes a rapid and efficient membrane fusion, in spite of no fusions with each peptide by itself. Each peptide takes nearly random coils with a small amount of helix, but the peptide mixture has an ordered helical structure. The equimolar peptide mixture forms a much more hydrophobic complex than those of different molar ratios of peptides and also that of each peptide itself. The equimolar peptide mixture causes the most efficient fusion. Preincubations of two peptides before addition to vesicles cause the slower rates of fusion. The fusion is greatly reduced at higher ionic strength and nearly zero at 800 mM NaCl and 40 mM sodium phosphate. Each peptide and the peptide mixture show the same alpha-helical structure, interact with vesicles, but do not induce fusion at higher ionic strengths. These results suggest that the two peptides interact mutually through the electrostatic Coulombic interaction between the charged groups. The electrically neutralized hydrophobic complex aggregates the separate vesicles together and interacts with the hydrocarbon region of lipid bilayers to cause fusion.     

Effects of composition and molecular packing on ionic properties of lipid monolayer having both cationic and anionic head groups

The effects of composition and molecular packing on the overall ionic property of mixed monolayer involving cationic, anionic and zwitterionic lipids were studied by measuring surface potential change when the concentration of sodium or calcium ions in the aqueous substrate was varied. Those lipids used were N,N-dimethyl-N-n-hexadecyl-n-octadecyl ammonium chloride (DDAC) as cationic lipid, stearic acid (StH) or sodium docosylsulfate (SDocS) as anionic lipid, and 1,2-dipalmitoyl-sn-glycero-3-phosphorylethanolamine (PE) and 1,2-distearoyl-sn-glycero-3-phosphorylcholine (PC) as ampholytic lipids. For the equimolar mixture of StH and DDAC, the reversal of apparent charge is observed when molecular packing exceeds 3.25 X 10(-10) M/cm2. The effect is rendered to the discreteness of cationic and anionic charges in the monolayer. It was also found that the addition of 30% of PC drastically changes the ionic property of PE closer to that of PC.     

Structural diversity in five-coordinate nickel(II) complexes of the tripyrrin ligand

Three different five coordinate nickel(II) complexes of tripyrrin ligands with chloro, oxalato and nitrato anionic ligands were obtained by ligand exchange reactions from respective trifluoroacetato species prepared in situ. Crystallographic studies of these compounds revealed different coordination geometries as well as different packing pattern. In the solid, the chloride complex accepts one water ligand to form a distorted trigonal bipyramid with two N donor centers in apical and one in an equatorial position. The molecules are organized in the crystal via hydrogen bonds, resulting in endless chains. Oxalate serves as a bridging ligand between two nickel(II) tripyrrins. Again the coordination of nickel(II) is found to be trigonal bipyramidal but with two equatorial and one apical nitrogen donors. The discrete dinuclear complexes are arranged in the crystal in a way as to form channels filled with toluene molecules. The nitrate species displays a η² bound nitrate ligand and short contacts between the nickel(II) center and an ethyl substituent of a neighboring molecule. The complex shows an unusually distorted molecular structure and unexpected differences in the two Ni-O bond lengths.     

Fluorescence quenching of beta-carboline alkaloids in micellar media. A study to select the adequate surfactant to use in analytical techniques.

The study of fluorescence quenching of the fluorophores allows the localization of the alkaloids (harmane and harmine) in the micelles (SDS, CTAB, Brij-35) to be established. In aqueous micellar solutions (SDS and Brij-35) at pH 13.0, emission corresponding to the neutral or zwitterionic forms can be observed. In the presence of CTAB (pH = 13.0) it was possible to observe the emission of anionic form. These species are not present in buffered aqueous solutions at these pH values. Bromide ion was added to the different surfactant solutions and the quenching effect was studied according to the Stern-Volmer equation. In the presence of SDS the quenching effect is considerably reduced compared to the aqueous solutions without surfactants, while for Brij-35 micelles were similar to those observed in homogeneous aqueous solution. For CTAB micelles a notable fluorescence quenching was observed for the different pH values studied. The fluorescence quenching studies show that the neutral species are associated inside the micelles, instead of the ionic species (cationic, zwitterionic or anionic) remaining on the surface of the micelles. The anionic surface of SDS micelles prevents the quenching effect by anionic quenchers for both neutral and charged species.     

Peroxyoxalate chemiluminescence enhanced by oligophenylenevinylene fluorophores in the presence of various surfactants

The effect of several surfactants on peroxyoxalate chemiluminescence (PO‐CL) using oligophenylenevinylene fluorophores was investigated. Among several oligophenylenevinylenes consisting of stilbene units, linearly conjugated ones, such as distyrylbenzene and distyrylstilbene, effectively enhanced PO‐CL efficiency. Various effects of anionic, cationic, amphoteric and non‐ionic surfactants on the CL efficiency of PO‐CL were determined using three oxalates and the distyrylbenzene fluorophore. Anionic and non‐ionic surfactants effectively enhanced CL efficiency, in contrast to the negative effect of cationic and amphoteric surfactants. Non‐ionic surfactants were also effective in CL reactions of oxalates bearing dodecyl ester groups by the hydrophobic interaction between their alkyl chains. Considering these results, the surfactants not only increase the concentrations of water‐insoluble interacting species in the hydrophobic micelle cores, but also control rapid degradation of the oxalates by alkaline hydrolysis. Copyright © 2014 John Wiley & Sons, Ltd.     

The influence of pH on cystine and dibasic amino acid transport by rat renal brushborder membrane vesicles

The uptake of cystine and lysine by rat renal brushborder membrane vesicles was examined at various intravesicular and extravesicular hydrogen ion concentrations to discern whether ionic species are determinants of specificity for the shared transport system and whether hydrogen ion gradients play a role in determining uptake values. When intravesicular and extravesicular pH are identical, the highest uptake of cystine occurred at pH 7.4, with lesser uptake at pH 6.0 and 8.3. Since cystine is electroneutral at pH 6.0 and 90% anionic at pH 8.3, it appears that neither form of the amino acid is a preferred species for transport. A similar relationship between pH and uptake occurs for lysine, which is cationic at pH below 8.5. This suggests that pH affects the functioning of the membrane carrier system independent of ionic species of the substrate. There is no apparent relationship of cystine uptake to hydrogen ion gradients across the membrane. Over the range of extravesicular pH studied, optimal cystine uptake occurred whenever the intravesicular pH was 7.4. Competitive interactions between unlabeled amino acids and labeled cystine were not affected by the extravesicular pH and, therefore, did not seem determined by the ionic species of cystine.     

Chitosan-colloidal gold complexes as polycationic probes for the detection of anionic sites by transmission and scanning electron microscopy

A new cationic colloidal gold complex has been developed for ultrastructural localization of cell surface anionic sites by transmission and scanning electron microscopy. The marker is prepared by labelling gold particles of suitable sizes (6 to 70 nm in diameter) with chitosan, a polymer of beta (1----4)-linked D-glucosamine. Using human red blood cells as a model, chitosan-gold complexes were shown to be specific for anionic sites and at pH 2 for sialic acid residues. The binding capacity of complexes of different sizes with carboxymethyl and phosphorylated celluloses was examined as a function of pH and ionic strength. The results indicated that these complexes can be used under acidic conditions as well as in physiological buffers. The complexes were further tested by transmission and scanning electron microscopy in detecting anionic sites on cells of various origins such as Escherichia coli, Lactobacillus maltaromicus, Lactobacillus reuteri, Saccharomyces cerevisiae, Saccharomyces rouxii, Schizosaccharomyces pombe, Fusarium oxysporum, Catharantus roseus.