Poly(vinyl amine)-silica composite nanoparticles: models of the silicic acid cytoplasmic pool and as a silica precursor for composite materials formation

The role of polymer (poly(vinylamine)) size (238-11000 units) on silicic acid condensation to yield soluble nanoparticles or composite precipitates has been explored by a combination of light scattering (static and dynamic), laser ablation combined with aerosol spectrometry, IR spectroscopy, and electron microscopy. Soluble nanoparticles or composite precipitates are formed according to the degree of polymerization of the organic polymer and pH. Nanoparticles prepared in the presence of the highest molecular weight polymers have core-shell like structures with dense silica cores. Composite particles formed in the presence of polymers with extent of polymerization below 1000 consist of associates of several polymer-silica nanoparticles. The mechanism of stabilization of the "soluble" silica particles in the tens of nanometer size range involves cooperative interactions with the polymer chains which varies according to chain length and pH. An example of the use of such polymer-poly(silicic acid) nanoparticles in the generation of composite polymeric materials is presented. The results obtained have relevance to the biomimetic design of new composite materials based on silica and polymers and to increasing our understanding of how silica may be manipulated (stored) in the biological environment prior to the formation of stable mineralized structures. We suspect that a similar method of storing silicic acid in an active state is used in silicifying organisms, at least in diatom algae.     

Release of Si from Silicon,a Ferrosilicon (FeSi) Alloy and a Synthetic Silicate Mineral in Simulated Biological Media

Unique quantitative bioaccessibility data has been generated, and the influence of surface/material and test media characteristics on the elemental release process were assessed for silicon containing materials in specific synthetic body fluids at certain time periods at a fixed loading. The metal release test protocol, elaborated by the KTH team, has previously been used for classification, ranking, and screening of different alloys and metals. Time resolved elemental release of Si, Fe and Al from particles, sized less than 50 µm, of two grades of metallurgical silicon (high purity silicon, SiHG, low purity silicon, SiLG), an alloy (ferrosilicon, FeSi) and a mineral (aluminium silicate, AlSi) has been investigated in synthetic body fluids of varying pH, composition and complexation capacity, simple models of for example dermal contact and digestion scenarios. Individual methods for analysis of released Si (as silicic acid, Si(OH)₄) in synthetic body fluids using GF-AAS were developed for each fluid including optimisation of solution pH and graphite furnace parameters. The release of Si from the two metallurgical silicon grades was strongly dependent on both pH and media composition with the highest release in pH neutral media. No similar effect was observed for the FeSi alloy or the aluminium silicate mineral. Surface adsorption of phosphate and lactic acid were believed to hinder the release of Si whereas the presence of citric acid enhanced the release as a result of surface complexation. An increased presence of Al and Fe in the material (low purity metalloid, alloy or mineral) resulted in a reduced release of Si in pH neutral media. The release of Si was enhanced for all materials with Al at their outermost surface in acetic media.     

Silica in plants: biological, biochemical and chemical studies

BACKGROUND: The incorporation of silica within the plant cell wall has been well documented by botanists and materials scientists; however, the means by which plants are able to transport silicon and control its polymerization, together with the roles of silica in situ, are not fully understood. RECENT PROGRESS: Recent studies into the mechanisms by which silicification proceeds have identified the following: an energy-dependent Si transporter; Si as a biologically active element triggering natural defence mechanisms; and the means by which abiotic toxicities are alleviated by silica. A full understanding of silica formation in vivo still requires an elucidation of the role played by the environment in which silica formation occurs. Results from in-vitro studies of the effects of cell-wall components associated with polymerized silica on mineral formation illustrate the interactions occurring between the biomolecules and silica, and the effects their presence has on the mineralized structures so formed. SCOPE: This Botanical Briefing describes the uptake, storage and function of Si, and discusses the role biomolecules play when incorporated into model systems of silica polymerization as well as future directions for research in this field.     

Microbially mediated aluminosilicate formation in acidic anaerobic environments: A cell‐scale chemical perspective

Through the use of scanning transmission electron microscopy (STEM) combined with other complementary techniques (SEM, cryo‐TEM, HRTEM, and EELS), we have studied the interaction of microorganisms inhabiting deep anoxic waters of acidic pit lakes with dissolved aluminum, silica, sulfate, and ferrous iron. These elements were close to saturation (Al, SiO₂) or present at very high concentrations (0.12 m Fe(II), 0.12–0.22 m SO₄^2?) in the studied systems. The anaerobic conditions of these environments allowed investigation of geomicrobial interactions that are difficult to see in oxidized, Fe(III)‐rich environments. Detailed chemical maps and through‐cell line scans suggest both extra‐ and intracellular accumulation of Al, Si, S, and Fe(II) in rod‐like cells and other structures (e.g., spherical particles and bacteriomorphs) of probable microbial origin. The bacterial rods showed external nanometric coatings of adsorbed Fe(II) and Al on the cell surface and cell interiors with significant presence of Al, Si, and S. These microbial cells coexist with spherical particles showing similar configuration (Fe(II) external coatings and [Al, Si, S]‐rich cores). The Al:Si and Al:S ratios and the good Al–Si correlation in the cell interiors suggest the concurrent formation of two amorphous phases, namely a proto‐aluminosilicate with imogolite‐like composition and proto‐hydrobasaluminite. In both cases, the mineralization appears to comprise two stages: a first stage of aluminosilicate and Al‐hydroxysulfate precipitation within the cell or around cellular exudates, and a second stage of SO₄^2? and Fe(II) adsorption on surface sites existing on the mineral phases in the case of (SO₄^2?) or on presumed organic molecules [in the case of Fe(II)]. These microbially related solids could have been formed by permineralization and mineral replacement of senescent microbial cells. However, these features could also denote biomineralization by active bacterial cells as a detoxification mechanism, a possibility which should be further explored. We discuss the significance of the observed Al/microbe and Si/microbe interactions and the implications for clay mineral formation at low pH.     

Silicic acid (Si(OH)(4)) is a significant influence upon the atomic absorption signal of aluminium measured by graphite furnace atomic absorption spectrometry (GFAAS).

We have identified silicic acid (Si(OH)(4)) as an important modifier of the absorbance signal of aluminium measured by graphite furnace atomic absorption spectrometry (GFAAS). The presence of Si(OH)(4) enhanced the signal by as much as 50%. The extent of the enhancement was dependent upon both [Al] and [Si(OH)(4)] and was maximal when [Al]< or =4.44 micromol dm(-3) and [Si(OH)(4)]> or =0.50 mmol dm(-3). The enhancement of the Al absorbance signal was not linearly related to [Si(OH)(4)] and the effect was, generally, saturated, for all [Al] tested, at [Si(OH)(4)]> or =0.50 mmol dm(-3). Si(OH)(4) was significantly more effective in enhancing the Al absorbance signal than Mg(NO(3))(2). However, the co-occurrence of 10 mmol dm(-3) Mg(NO(3))(2) and 2 mmol dm(-3) Si(OH)(4) in samples abolished the enhancement due to Si(OH)(4). The presence of Si(OH)(4) in samples could result in an overestimation of the Al content of those samples by as much as 50%. Errors in the measurement of Al in samples containing Si(OH)(4) could be prevented using matrix-matched calibration standards. Our observation could have serious implications for the determination of Al in aqueous samples of both geochemical and biological interest. It may also point towards the application of Si(OH)(4) as a novel and effective matrix modifier in the determination of Al by GFAAS since the inclusion of Si(OH)(4) in standards and samples improved the limit of detection of Al from ca 8 nmol dm(-3) to 3 nmol dm(-3).     

The use of root growth and modelling data to investigate amelioration of aluminium toxicity by silicon in Picea abies seedlings

Three-week-old Picea abies seedlings were grown for 7 days in 100 microM aluminium (Al), combined with 1000 or 2000 microM silicon (Si). Solution pH was adjusted to 4.00, 4.25, 4.50, 4.75, or 5.00. In the absence of Si, solution pH had no effect on the decrease in root growth caused by 100 microM Al. Silicon did not ameliorate toxic effects of Al on root growth at pH 4.00, 4.25 and 4.50, whereas significant, and apparently complete, amelioration was found at pH 4.75 and 5.00. An equilibrium speciation model (EQ3NR), with a current thermodynamic database, was used to predict the behaviour of Al and Si in growth solutions. When Si was not present in the 100 microM Al solutions, Al(3+) declined from 92.4% of total Al at pH 4.00 to 54.6% at pH 5.00, and there was a concomitant increase in hydroxyaluminium species as pH increased. The addition of 1000 microM Si to the 100 microM Al solutions caused a reduction in Al(3+) content over the whole pH range: at pH 4.00 Al(3+) fell from 92.4 to 83.3% in the presence of Si; and at pH 5.00 the fall was from 54.6 to 17.7%. These falls were attributed to the formation of hydroxyaluminosilicate (HAS) species. Similar, but somewhat greater, changes were observed in solutions containing 2000 microM Si. The match between root growth observations and the modelling data was not very good. Modelling predicted that change in Al(3+) content with pH in the presence of Si was gradual, but root growth was markedly increased between pH 4.50 and 4.75. Differences between root growth and modelling data may be due to the model not correctly predicting solution chemistry or to in planta effects which override the influence of solution chemistry.     

Multiparametric Analyses Reveal the pH-Dependence of Silicon Biomineralization in Diatoms

Diatoms, the major contributors of the global biogenic silica cycle in modern oceans, account for about 40% of global marine primary productivity. They are an important component of the biological pump in the ocean, and their assemblage can be used as useful climate proxies; it is therefore critical to better understand the changes induced by environmental pH on their physiology, silicification capability and morphology. Here, we show that external pH influences cell growth of the ubiquitous diatom Thalassiosira weissflogii, and modifies intracellular silicic acid and biogenic silica contents per cell. Measurements at the single-cell level reveal that extracellular pH modifications lead to intracellular acidosis. To further understand how variations of the acid-base balance affect silicon metabolism and theca formation, we developed novel imaging techniques to measure the dynamics of valve formation. We demonstrate that the kinetics of valve morphogenesis, at least in the early stages, depends on pH. Analytical modeling results suggest that acidic conditions alter the dynamics of the expansion of the vesicles within which silica polymerization occurs, and probably its internal pH. Morphological analysis of valve patterns reveals that acidification also reduces the dimension of the nanometric pores present on the valves, and concurrently overall valve porosity. Variations in the valve silica network seem to be more correlated to the dynamics and the regulation of the morphogenesis process than the silicon incorporation rate. These multiparametric analyses from single-cell to cell-population levels demonstrate that several higher-level processes are sensitive to the acid-base balance in diatoms, and its regulation is a key factor for the control of pattern formation and silicon metabolism.     

Analytical studies on the incorporation of aluminium in the cell walls of the marine diatom Stephanopyxis turris

The eukaryotic diatoms are unicellular algae. They are well known for their filigree micro- and nanostructured cell walls which mainly consist of amorphous silica as well as various organic compounds. However, diatoms are also known to incorporate certain amounts of aluminium into their cell walls. Unexpectedly, enhanced Al concentrations in the Southern Yellow Sea were found to be correlated with a diatom spring bloom. Therefore, we have analyzed the influence of strongly enhanced Al concentrations in the culture medium upon the growth behaviour of the diatom Stephanopyxis turris (S. turris). The uptake and incorporation of Al into the cell walls was monitored. It turned out that S. turris survives aluminium concentrations up to 105.5 μM (2.85 mg/l) in the culture medium. Under the applied conditions, this corresponds to an Al/Si ratio of 1:1. These large amounts of Al had to be offered in the form of bis–tris-chelates in order to prevent uncontrolled precipitation. Under these conditions, the Al/Si ratio in the cell walls could be increased up to about 1:15 as determined by ICP-OES, the highest amount of aluminium found in diatom cell walls yet. Structural characterization of the biosilica by ATR-FTIR and solid-state ²⁷Al NMR spectroscopy revealed that an amorphous aluminosilicate phase is formed where the aluminium exists as four- and sixfold-coordinated species.     

Effect of temperature,Si/Al and metal loading on the positioning of Pt atoms in MOR-type zeolites

Metropolis Monte Carlo (MMC) and molecular dynamics (MD) simulations were performed to study the feasibility of synthesising Pt nanowires inside the pores of zeolites with the MOR framework, such as mordenite. Results show that the temperature, the silicon to aluminium ratio (Si/Al) of the zeolite and the Pt metal loading have an important combined effect on the positioning of Pt atoms inside the framework. MMC simulations indicate that low Si/Al and high metal loadings promote the positioning of Pt atoms inside the main pore channels, which are the places where metal nanowires could be formed. On the other hand, high Si/Al and low metal loadings favour the positioning of Pt atoms in the side pockets of the MOR structure. For Pt loadings of 5% (mol/mol), the guest metal atoms were positioned mainly in the main pore channels for all conditions studied. MD simulation results are in agreement with MMC simulations, showing that when Si/Al = ∞, the trend of single Pt atoms is to move towards the side pockets of the MOR framework, while for Si/Al = 5, the Pt atoms remain in the main pore channel of the structure. Temperature had the effect of increasing the mobility of Pt atoms through the MOR framework.     

Silicon amelioration of aluminium toxicity and cell death in suspension cultures of Norway spruce (Picea abies (L.) Karst.)

A role for silicon (Si) in the amelioration of aluminium (Al) toxicity in gymnosperms is suggested by their codeposition in planta, including within needles. This study was designed to investigate Al/Si interactions at the cellular level using suspension cultures of Norway spruce. Toxic effects of Al were dependent on duration of Al exposure, concentration of Al, and pH. Toxicity was reduced when Si was present, and the effect was enhanced at pH 5.0 compared to pH 4.2. Study of the ultrastructure of Al-treated cells indicated that changes in cell wall thickening, degree of vacuolation, and the degeneration of mitochondria, Golgi bodies, ER and nucleus preceded cell death, and significant amelioration was noted when Si was also present. When the fluorescent dye Morin was employed to localise free Al, cells treated with Al and Si in combination showed less fluorescence than the cells treated with Al alone. Intensity of fluorescence depended on the concentration of Al, duration of treatment and pH. Notably, presence of Si reduced the concentration of free Al in the cell wall in parallel with amelioration of Al toxicity. We therefore propose that formation of aluminosilicate complexes in the wall and apoplasm provide a significant barrier to Al penetration and cell damage in Norway spruce.     

Ultrastructural evidence for uptake of silicon-containing silicic acid analogs byUrtica pilulifera and incorporation into cell wall silica

Summary In natural environments the stinging nettle plant,Urtica pilulifera, bears stinging cells in which electron dense silica deposits occupy a significant volume of the cell wall. Plants were grown in hydroponic solutions with and without supplements of silicic acid, the chemical form of silicon available to biological systems to determine if this plant and the stinging cells will grow normally under conditions of silicon starvation. In separate experiments, several analogs of silicic acid were added as supplements to the hydroponic solution to determine whether silicic acid binding sites had detectably different specificities for the different molecular structures of the analogs. The analogs [(R-)_nSi(-OH)_m] have the following structures (R, n, m): (1)-H, 1, 3; (2)-CH₃, 1, 3; (3)-CH₃, 2, 2; (4)-CH₃, 3, 1; (5)-CH₂CH₃, 1, 3; and (6)-C₆H₅, 1, 3. Electron microscopy was used as an assay for the uptake and incorporation of analogs into an electron dense silica-like product in the stinging cell wall. The results indicate that cell wall silica production occurred only when the analog contained at least three hydroxyl groups. The morphology and ontogeny of the plant was normal except for: 1, the appearance of green spots on the leaves when the analog contained two or more hydroxyl groups, and 2, total blockage of flowering by the two methyl derivative of silicic acid, (CH₃)₂Si(OH)₂.     

Direct and indirect identification of the formation of hydroxyaluminosilicates in acidic solutions.

Morin-aluminium fluorescence and membrane filtration were successfully applied to the indirect identification of the formation of hydroxyaluminosilicates (HAS) in acidic solutions of varying pH and of known concentrations of aluminium (Al) and silicic acid (Si(OH)(4)). It was proven to be especially useful in providing evidence of the strong competition between Si(OH)(4) and Al(OH)(3) to condense with hydroxyaluminium templates to form HAS in preference to Al(OH)(3(s)). The aggregation and stability of HAS and Al(OH)(3(s)) were dependent upon both the pH and the [Al] of the solution. The applicability of these indirect techniques was confirmed using the direct observation of HAS in solution by atomic force microscopy (AFM). AFM was also a powerful tool in providing valuable information on the morphology of colloidal HAS of various structures and stoichiometries. The results have provided further confirmation of both the mechanism of HAS formation and the form and stability of HAS in solution. This information is essential to our understanding of the biological availability and hence toxicity of Al in biota, including man.     

Expression, purification, and reconstitution of a diatom silicon transporter

The synthesis and manipulation of silicon materials on the nanoscale are core themes in nanotechnology research. Inspiration is increasingly being taken from the natural world because the biological mineralization of silicon results in precisely controlled, complex silica structures with dimensions from the millimeter to the nanometer. One fascinating example of silicon biomineralization occurs in the diatoms, unicellular algae that sheath themselves in an ornate silica-based cell wall. To harvest silicon from the environment, diatoms have developed a unique family of integral membrane proteins that bind to a soluble form of silica, silicic acid, and transport it across the cell membrane to the cell interior. These are the first proteins shown to directly interact with silicon, but the current understanding of these specific silicon transport proteins is limited by the lack of in vitro studies of structure and function. We report here the recombinant expression, purification, and reconstitution of a silicon transporter from the model diatom Thalassiosira pseudonana. After using GFP fusions to optimize expression and purification protocols, a His(10)-tagged construct was expressed in Saccharomyces cerevisiae, solubilized in the detergent Fos-choline-12, and purified by affinity chromatography. Size-exclusion chromatography and particle sizing by dynamic light scattering showed that the protein was purified as a homotetramer, although nonspecific oligomerization occurred at high protein concentrations. Circular dichroism measurements confirmed sequence-based predictions that silicon transporters are α-helical membrane proteins. Silicic acid transport could be established in reconstituted proteoliposomes, and silicon uptake was found to be dependent upon an applied sodium gradient. Transport data across different substrate concentrations were best fit to the sigmoidal Hill equation, with a K(0.5) of 19.4 ± 1.3 μM and a cooperativity coefficient of 1.6. Sodium binding was noncooperative with a K(m)(app) of 1.7 ± 1.0 mM, suggesting a transport silicic acid:Na(+) stoichiometry of 2:1. These results provide the basis for a full understanding of both silicon transport in the diatom and protein-silicon interactions in general.     

Cell surface elemental composition of Microcystis aeruginosa: high-Si and low-Si subpopulations within the water column of a eutrophic lake

Energy-dispersive X-ray microanalysis (10 kV microprobe) wascarried out to determine the surface elemental composition ofsingle cells of Microcystis aeruginosa within mixed phytoplanktonpreparations, sampled at different depths (0–8 m) withina stratified eutrophic lake. Mean elemental concentrations (mmolkg^–1 dry weight) throughout the sampled water column were:magnesium, 125; silicon (Si), 1864; phosphorus, 341; sulphur,122; chlorine, 88; potassium, 282; calcium, 63. Although somesignificant differences in elemental composition occurred withdepth, the underlying pattern was one of relative uniformitywithin the top 8 m of the water column. At each depth, the frequencydistribution of Si was bimodal, indicating two distinct subpopulationsof cells with cell surface Si concentrations ranging from 0to 1000 mmol kg^–1 (overall mean 112, low-Si cells) andfrom 1000 to 6800 mmol kg^–1 (overall mean 3649, high-Sicells). The two subpopulations occurred mixed together withinindividual colonies, and both included dividing and non-dividingcells. Both cell types had a wide range of sizes, but high-Sicells reached a higher maximum size in both dividing (diametergreater by 0.90 µm) and non-dividing (difference of 0.44µm) cells. This difference in size is consistent withSi being present as a surface layer (up to 0.2–0.4 µmin thickness) in high-Si cells. Support for the presence ofa surface layer of Si [with aluminium (Al)] is also providedby correlation analysis (Si and Al are significantly negativelycorrelated with other cell elements), principal component analysis(Si and Al occur as a distinct subgroup) and lower mean concentrationsof elements (other than Al) in high-Si cells (due to reducedX-ray contribution from the cell interior). The proportion ofhigh-Si cells in the water column was locally high in the 2m depth sample, and had an overall value of 40.5% for the wholedata set. The biological significance of high- and low-Si cellsis not known.     

Putative silicon transport vesicles in the cytoplasm of the diatom Synedra acus during surge uptake of silicon

We studied the growth of the araphid pennate diatom Synedra acus subsp. radians (Kützing) Skabichevskii using a fluorescent dye N ¹,N ³-dimethyl-N ¹-(7-nitro-2,1,3-benzoxadiazol-4-yl)propane-1,3-diamine (NBD-N2), which stains growing siliceous frustules but does not stain other subcellular organelles. We used a clonal culture of S. acus that was synchronized by silicon starvation. Epifluorescence microscopy was performed in two different ways with cells stained by the addition of silicic acid and the dye. Individual cells immobilized on glass were observed during the first 15–20 min following the replenishment of silicic acid after silicon starvation. Alternatively, we examined cells of a batch culture at time intervals during 36 h after the replenishment of silicic acid using fluorescence and confocal microscopy. The addition of silicic acid and NBD-N2 resulted in the rapid (1–2 min) formation of several dozen green fluorescent submicrometer particles (GFSPs) in the cytoplasm, which was accompanied by the accumulation of fluorescent silica inside silica deposition vesicles (SDVs) along their full length. In 5–15 min, GFSPs disappeared from the cytoplasm. Mature siliceous valves were formed within the SDVs during the subsequent 14–16 h. In the next 8–10 h, GFSPs appeared again in the cytoplasm of daughter cells. The data obtained confirm observations about the two-stage mechanism of silicon assimilation, which includes rapid silicon uptake (surge uptake) followed by slow silica deposition. It is likely that the observed GFSPs are silicon transport vesicles, which were first proposed by Schmid and Schulz in (Protoplasma 100:267–288, 1979).     

Effects of germanium on the morphology of silica deposition in a freshwater sponge

The effect of germanium on the secretion of siliceous spicules by the freshwater sponge Spongilla lacustris was investigated by exposing germinating and hatching gemmules to varying concentrations of germanium (Ge) in the presence of silicon (Si). Results were analyzed quantitatively and qualitatively and demonstrate that a [Ge]/[Si] (= molar ratio) of 1.0 completely inhibits silicon deposition. Intermediate ratios (0.5, 0.1, 0.01) which are permissive to spicule appearance result in fewer, shorter, and thinner spicules, in proportionately fewer microscleres, and in short bulbous megascleres. The size of the bulb increases with increasing [Ge]/[Si], while the length of the bulbous megascleres decreases with increasing [Ge]/[Si]. Microscleres do not demonstrate these graded responses suggesting that they are secreted in an all or none manner. Swellings produced in pond water and bulbs produced in germanium appear to decrease in size with time indicating a spreading of the accumulated silica. The effect of germanium on spicule secretion can be partially explained by its ability to uncouple the growth in length of the axial filament from the growth of the surrounding silicalemma. Under these conditions excess silicalemma is produced in which silica accumulates as bulbs in short spicules. Continuous exposure to Ge is necessary to produce this altered morphology. It is conjectured that the bulbs may be retained due to an inhibition of spreading. which in turn may be caused by the incorporation of germanium into the silica.     

Electron micrographic study of precipitates formed by interaction of silicic acid and alkaline phosphatase: contribution to a study of silica urolithiasis in cattle

Association of alkaline phosphatase with silicic acid in precipitates formed in dilute solution was studied as a model for the nonspecific reaction between silicic acid and protein. Precipitates contained 68-83% of the silicic acid and 52-83% of the enzyme in the original mixture and were in the form of aggregates of roundish particles 150-800 nm in diameter. Enzyme protein formed a tightly bound layer on the surface of particles formed in solutions of freshly prepared silicic acid. The similarity between the ultrastructural features of precipitates from solutions of silicic acid and of internal portions of siliceous urinary calculi from cattle suggests that deposition of silica during development of such calculi is due, at least in part, to the interaction of protein with silicic acid in urine.     

Effects of Germanium (Ge) on the silica spicules of the marine sponge Suberites domuncula: Transformation of spicule type

Germanium (Ge), in the form of germanic acid, at a Ge/Si molar ratio of 1.0 inhibits gemmule development and silica deposition in the marine demosponge Suberites domuncula. Lower Ge/Si ratios inhibit the growth in length of the silica spicules (tylostyles) producing short structures, but with relatively normal morphology and close to normal width; spherical protuberances occasionally occur on these spicules. A few of the short spicules possess completely round rather than pointed tips. Many of the latter develop when Ge is added (pulsed) to growing animals, thus inducing a change in spicule type. These results indicate that the growth in length of the axial filament is more sensitive to Ge inhibition than is silica deposition and that pointed spicule tips normally develop because the growth of the axial filament at the spicule tip is more rapid than silica deposition. Newly formed spicules initiate silica deposition at the spicule head but the absence of Ge-induced bulbs as in freshwater spicules (oxeas) leaves open the question of whether there is a silicification center(s) present in Suberites tylostyles. The morphogenesis of freshwater oxeas and of marine tyolstyles appears fundamentally different-bidirectional growth in the former and unidirectional growth in the latter. X-ray analysis demonstrate relatively uniform Ge incorporation into the silica spicules with considerable variation from spicule to spicule in the incorporated level. Increased silicic acid concentration induces the formation of siliceous spheres, suggesting that the axial filament becomes prematurely encased in silica.     

Regular consumption of a silicic acid-rich water prevents aluminium-induced alterations of nitrergic neurons in mouse brain: histochemical and immunohistochemical studies

Silicon is not generally considered an essential nutrient for mammals and, to date, whether it has a biological role or beneficial effects in humans is not known. The results of a number of studies suggest that dietary silicon supplementation might have a protective effect both for limiting aluminium absorption across the gut and for the removal of systemic aluminium via the urine, hence, preventing potential accumulation of aluminium in the brain. Since our previous studies demonstrated that aluminium exposure reduces the number of nitrergic neurons, the aim of the present study was to compare the distribution and the morphology of NO-containing neurons in brain cortex of mice exposed to aluminium sulphate dissolved in silicic acid-rich or poor drinking water to assess the potential protective role of silicon against aluminium toxicity in the brain. NADPH-d histochemistry and nNOS immunohistochemistry showed that high concentrations of silicon in drinking water were able to minimize the impairment of the function of nitrergic neurons induced by aluminium administration. We found that silicon protected against aluminium-induced damage to the nitrergic system: in particular, we demonstrated that silicon maintains the number of nitrergic neurons and their expression of nitrergic enzymes at physiological levels, even after a 12 and 15 month exposure to aluminium.     

The Interaction Between Silicon and Aluminium in Sorghum bicolor (L.) Moench: Growth Analysis and X-ray Microanalysis

Seeds of Sorghum bicolor (L.) Moench. were germinated on moistfilter paper for 6 d, before the seedlings were transferredto pots containing 500 µmol l^-1 Ca(NO₃)₂ for 2 d. Theseedlings were then treated with 0 or 100 µmol l^-1 Alin factorial combination with 0, 1400 or 2800 µmol l^-1Si for 8 d. The background solution used throughout was 500µmol l^-1 Ca(NO₃)₂. Aluminium treatment reduced root growthand caused a significant increase in shoot/root ratio. The presenceof silica in the solution significantly ameliorated the effectsof aluminium on root growth. Three treatment were selected for a microanalytical investigationof the basal region of the root: 2800 µmol l^-1 Si only;100 µmol l^-1 Al only; and a combination of the two. Inthe 2800 µmol l^-1 treatment silica was deposited in theendodermis, with the greatest accumulation being in the innertangential wall (ITW). When plants were treated with 100 µmoll^-1 Al only, aluminium concentration was highest in the outertangential wall (OTW) of the epidermis. The element was presentin the hypodermal walls and OTW of the endodermis and was notdetectable in the stele. With both 2800 µmol l^-1 Si and100 µmol l^-1 Al in the nutrient solution the two biomineralizationsites were the ITW of the endodermis, where silicon was themajor element deposited, and atypically in the OTW of the epidermiswhere both aluminium and silicon were present. The sequestrationof aluminium in the Al-Si deposit in the OTW of the epidermismay represent the mechanism that allows greater root growthin this treatment.Copyright 1993, 1999 Academic Press Sorghum bicolor (L.) Moench., aluminium, silicon, calcium, root, toxicity, biomineralization, X-ray microanalysis, freeze substitution