Effects of copper, and aluminium in ionic, and nanoparticulate form on growth rate and gene expression of Setaria italica seedlings.

This study aims to determine the effects of copper, copper oxide nanoparticles, aluminium, and aluminium oxide nanoparticles on the growth rate and expression of ACT-1, CDPK, LIP, NFC, P5CR, P5CS, GR, and SiZIP1 genes in five days old seedling of Setaria italica ssp. maxima, cultivated in hydroponic culture. Depending on their concentration (ranging from 0.1 to 1.8 mg L-1), all tested substances had both stimulating and inhibiting effects on the growth rate of the seedlings. Copper and copper oxide-NPs had generally a stimulating effect whereas aluminium and aluminium oxide-NPs at first had a positive effect but in higher concentrations they inhibited the growth. Treating the seedlings with 0.4 mg L-1 of each tested toxicant was mostly stimulating to the expression of the genes and reduced the differences between the transcript levels of the coleoptiles and roots. Increasing concentrations of the tested substances had both stimulating and inhibiting effects on the expression levels of the genes. The highest expression levels were usually noted at concentrations between 0.4 and 1.0 mg/L of each metal and metal nanoparticle, except for SiZIP1, which had the highest transcript amount at 1.6 mg L-1 of Cu2+ and at 0.1-0.8 mg L-1 of CuO-NPs, and LIP and GR from the seedling treated with Al2O3-NPs at concentrations of 0.1 and 1.6 mg L-1, respectively.

Highly Crystalline Copper Aluminum-Layered Double Hydroxides with Intrinsic Fenton-Like Catalytic Activity for Robust Oral Health Management.

As the main challenge of dental healthcare, oral infectious diseases are highly associated with the colonization of pathogenic microbes. However, current antibacterial treatments in the field of stomatology still lack a facile, safe, and universal approach. Herein, we report the controllable synthesis of copper aluminum-layered double hydroxides (CuAl-LDHs) with high Fenton-like catalytic activity, which can be utilized in the treatment of oral infectious diseases with negligible side effects. Our strategy can efficiently avoid the unwanted doping of other divalent metal ions in the synthesis of Cu-contained LDHs and result in the formation of binary CuAl-LDHs with high crystallinity and purity. Evidenced by experimental and theoretical results, CuAl-LDHs exhibit excellent catalytic ability toward the ·OH generation in the presence of H2O2 and hold strong affinity toward bacteria, endowing them with great catalytic sterilization against both Gram-positive and Gram-negative bacteria. As expected, these CuAl-LDHs provide outstanding treatments for mucosal infection and periodontitis by promoting wound healing and remodeling of the periodontal microenvironment. Moreover, toxicity investigation demonstrates the overall safety. Accordingly, the current study not only provides a convenient and economic strategy for treating oral infectious diseases but also extends the development of novel LDH-based Fenton or Fenton-like antibacterial reagents for further biomedical applications.

A Platinum-Aluminum Bimetallic Salen Complex for Pro-senescence Cancer Therapy.

Cell senescence is defined as irreversible cell cycle arrest, which can be triggered by telomere shortening or by various types of genotoxic stress. Induction of senescence is emerging as a new strategy for the treatment of cancer, especially when sequentially combined with a second senolytic drug capable of killing the resulting senescent cells, however severely suffering from the undesired off-target side effects from the senolytic drugs. Here, we prepare a bimetalic platinum-aluminum salen complex (Alumiplatin) for cancer therapy-a combination of pro-senesence chemotherapy with in situ senotherapy to avoid the side effects. The aluminum salen moiety, as a G-quadruplex stabilizer, enhances the salen's ability to induce cancer cell senescence and this phenotype is in turn sensitive to the cytotoxic activity of the monofunctional platinum moiety. It exhibits an excellent capability for inducing senescence, a potent cytotoxic activity against cancer cells both in vitro and in vivo, and an improved safety profile compared to cisplatin. Therefore, Alumiplatin may be a good candidate to be further developed into safe and effective anticancer agents. This novel combination of cell senescence inducers with genotoxic drugs revolutionizes the therapy options of designing multi-targeting anticancer agents to improve the efficacy of anticancer therapies.

Effect of TiC Nanoparticles on a Zn-Al-Cu System for Biodegradable Cardiovascular Stent Applications.

Despite being a weaker metal, zinc has become an increasingly popular candidate for biodegradable implant applications due to its suitable corrosion rate and biocompatibility. Previous studies have experimented with various alloy elements to improve the overall mechanical performance of pure Zn without compromising the corrosion performance and biocompatibility; however, the thermal stability of biodegradable Zn alloys has not been widely studied. In this study, TiC nanoparticles were introduced for the first time to a Zn-Al-Cu system. After hot rolling, TiC nanoparticles were uniformly distributed in the Zn matrix and effectively enabled phase control during solidification. The Zn-Cu phase, which was elongated and sharp in the reference alloy, became globular in the nanocomposite. The strength of the alloy, after introducing TiC nanoparticles, increased by 31% from 259.7 to 340.3 MPa, while its ductility remained high at 49.2% elongation to failure. Fatigue performance also improved greatly by adding TiC nanoparticles, increasing the fatigue limit by 47.6% from 44.7 to 66 MPa. Furthermore, TiC nanoparticles displayed excellent phase control capability during body-temperature aging. Without TiC restriction, Zn-Cu phases evolved into dendritic morphologies, and the Al-rich eutectic grew thicker at grain boundaries. However, both Zn-Cu and Al-rich eutectic phases remained relatively unchanged in shape and size in the nanocomposite. A combination of exceptional tensile properties, improved fatigue performance, better long-term stability with a suitable corrosion rate, and excellent biocompatibility makes this new Zn-Al-Cu-TiC material a promising candidate for biodegradable stents and other biodegradable applications.

Aluminum zirconate nanoparticles in etch and rinse adhesive to caries affected dentine: An in-vitro scanning electron microscopy, elemental distribution, antibacterial, degree of conversion and micro-tensile bond strength assessment.

To incorporate different concentrations of Al2O9Zr3 (1%, 5%, and 10%) nanoparticles (NP) into the ER adhesive and subsequently assess the impact of this addition on the degree of conversion, µTBS, and antimicrobial efficacy. The current research involved a wide-ranging examination that merged various investigative techniques, including the application of scanning electron microscopy (SEM) for surface characterization of NP coupled with energy-dispersive x-ray spectroscopy (EDX), Fourier-transform infrared (FTIR) spectroscopy, µTBS testing, and microbial analysis. Teeth were divided into four groups based on the application of modified and unmodified three-step ER adhesive primer. Group 1 (0% Al2O9Zr3 NPs) Control, Group 2 (1% Al2O9Zr3 NPs), Group 3 (5% Al2O9Zr3 NPs), and Group 4 (10% Al2O9Zr3 NPs). EDX analysis of Al2O9Zr3 NPs was performed showing elemental distribution in synthesized NPs. Zirconium (Zr), Aluminum (Al), and Oxides (O2). After primer application, an assessment of the survival rate of Streptococcus mutans was completed. The FTIR spectra were analyzed to observe the characteristic peaks indicating the conversion of double bonds, both before and after the curing process, for the adhesive Etch and rinse containing 1,5,10 wt% Al2O9Zr3 NPs. µTBS and failure mode assessment were performed using a Universal Testing Machine (UTM) and stereomicroscope respectively. The µTBS and S.mutans survival rates comparison among different groups was performed using one-way ANOVA and Tukey post hoc (p = .05). Group 4 (10 wt% Al2O9Zr3 NPs + ER adhesive) specimens exhibited the minimum survival of S.mutans (0.11 ± 0.02 CFU/mL). Nonetheless, Group 1 (0 wt% Al2O9Zr3 NPs + ER adhesive) displayed the maximum surviving S.mutans (0.52 ± 0.08 CFU/mL). Moreover, Group 2 (1 wt% Al2O9Zr3 NPs + ER adhesive) (21.22 ± 0.73 MPa) samples displayed highest µTBS. However, the bond strength was weakest in Group 1 (0 wt% Al2O9Zr3 NPs + ER adhesive) (14.13 ± 0.32 MPa) study samples. The etch-and-rinse adhesive exhibited enhanced antibacterial activity and micro-tensile bond strength (µTBS) when 1% Al2O9Zr3 NPs was incorporated, as opposed to the control group. Nevertheless, the incorporation of Al2O9Zr3 NPs led to a decrease in DC. RESEARCH HIGHLIGHTS: 10 wt% Al2O9Zr3 NPs + ER adhesive specimens exhibited the minimum survival of S.mutans. 1 wt% Al2O9Zr3 NPs + ER adhesive samples displayed the most strong composite/CAD bond. The highest DC was observed in Group 1: 0 wt% Al2O9Zr3 NPs + ER adhesive.

Insights into the Structure of Sucralfate by Advanced Solid- and Liquid-State NMR.

Sucralfate, which is a sucrose octasulfate aluminum complex, is an active pharmaceutical ingredient (API) falling in the category of cytoprotective agents which are very effective for gastric and duodenal ulcers. On interaction with stomach acid, it ionizes into aluminum and sucrose octasulfate ions to form a protective layer over the ulcerated region inhibiting further attack from acid. The mechanism of action of sucralfate in the context of its structure is not well understood. Considering that at least two forms of this API are available in the market, there are no reports on the various forms of sucralfate and differences in their pharmacological action. We characterized the two forms of sucralfate using multinuclear, multidimensional solid-state NMR, and the results show significant structural differences between them arising from variation in the aluminum environment and the level of hydration. The impact of structural differences on pharmacological action was examined by studying acid-induced Al release by 27Al liquid-state NMR. The sucralfate, European pharmaceutical standard, Form I, undergoes faster disruption in acid compared to Form II. The difference is explained on the basis of structural differences in the two forms which gives significant insights into the action of sucralfate in relation to its structure.

How do different concentrations of aluminum and zinc affect the survival, body size, morphology and immune system of Physalaemus cuvieri (Fitzinger, 1826) tadpole?

The assessment of amphibian responses as bioindicators of exposure to chemical pollutants is an important tool for conservation of native species. This study aimed to investigate the effects of chronic aluminum (Al) and zinc (Zn) exposure on survival, body size, morphology (malformations), and immune system (leukocyte profile) in P. cuvieri tadpoles. Ecotoxicological analyses were performed utilizing chronic toxicity tests in which 210 tadpoles at the 25th Gosner developmental stage were exposed to Al and Zn. Individuals of P. cuvieri were maintained in glass containers containing various concentrations of aluminum sulfate (0.1, 0.2, or 0.3 mg/L) and zinc sulfate (0.18, 0.27 or 0.35 mg/L), and tests were performed in triplicate. After 14 days, amphibians were weighed, measured and survival rate, malformations in the oral and intestine apparatus, leukocyte profile, and ratio between neutrophils and lymphocytes determined. The differing concentrations of Al and Zn did not produce lethality in P. cuvieri where 95% of the animals survived 326 hr following metal exposure. Individuals exposed to Zn achieved greater body growth and weight gain compared to controls. Aluminum increased weight gain compared controls. These metals also produced malformations of the oral and intestine apparatus and enhanced occurrence of hemorrhages, especially at the highest doses. Lymphocytes were the predominant cells among leukocytes, with lymphopenia and neutrophilia observed following Al and Zn treatment, as evidenced by elevated neutrophil/lymphocyte ratio, an important indicator of stress in animals. Data suggest that further studies need to be carried out, even with metal concentrations higher than those prescribed by CONAMA, to ensure the conservation of this species.

Biodegradable Monometallic Aluminum as a Biotuner for Tumor Pyroptosis.

Pyroptosis is an effective anti-tumor strategy. However, monometallic pyroptosis biotuners have not been explored until now. Here, we discover for the first time that biodegradable monometallic Al can act as a pyroptosis biotuner for tumor therapy. pH-sensitive Al nanoparticles (Al@P) are obtained by equipping polyethylene glycol-b-(poly(methyl methacrylate)-co-poly(4-vinylpyridine), which can exert their effect at the tumor site without affecting normal cells. The H2 and Al3+ release by Al@P in the acidic environment of tumors disrupts the redox balance and ionic homeostasis in tumor cells, thus generating large amounts of reactive oxygen species (ROS), leading to caspase-1 activation, gasdermin D cleavage, and IL-1ß/LDH release, which induces canonical pyroptotic death. Meanwhile, the prodrug Doxorubicin (Pro-DOX) is successfully loaded onto Al@P (Al@P-P) and can be activated by ROS to release DOX in the tumor cells, thus further improving the tumor-killing efficiency. Ultimately, Al@P-P is degradable and exhibits efficient tumor inhibition.

Difference in the cellular response following THP-1 derived phagocytic monocyte cells exposure to commercial aluminum-based adjuvants and aluminum-containing vaccines.

BACKGROUND: Aluminum-based adjuvants (ABAs) enhance the immune response following vaccine injection. Their mechanisms of action are not fully understood, and their bio-persistency have been described associated with long-term adverse effects. METHODS: We evaluated and compared the cellular effects of the two main ABAs and whole vaccines on ATP production, ROS generation and cytokines production (IL-6 and IL-10), using THP-1 cells. RESULTS: ABAs altered the cell energy metabolism by increasing ROS production after 24 h and reducing ATP production after 48 h. In addition, both ABAs and whole vaccines induced different kinetics of IL-6 production, whereas only ABAs induced IL-10 secretion. CONCLUSION: This study showed clearly, for a first time, a difference in cellular response to the ABAs and whole vaccines which should be taken into consideration in future studies focusing on the effect of ABA in vaccines. Future studies on ABAs should also pay attention to mitochondrial function alterations following exposure to ABA-containing vaccines.

Zinc/aluminum-layered double hydroxide-gallic acid doped carboxymethyl cellulose nanocomposite films for wound healing.

Effective loading and delivering the wound healing-based materials to the wound site and area with an optimum concentration and limited cytotoxicity are essential for a complete and fast healing process. Here, we have designed Zn/Al-LDH nanoparticles-loaded CMC films for encapsulation and delivery of gallic acid (GA) in order to develop an effective and efficient wound-healing scaffold. The physicochemical properties of the prepared Zn/Al-LDH nanohybrids were thoroughly characterized by several characterization techniques, such as FESEM, Hi-TEM, FTIR, and XRD techniques. The thermal properties of the scaffolds were evaluated by DSC and TGA analysis. The release profiles of GA from fabricated films were studied over 8 h by UV-vis spectroscopy. In vitro drug release studies in PBS solutions with pH 7.4 showed a mono-phasic profile in which the liberation of the drug mainly occurred by scaffold erosion and increased by increasing the experiment period. The in vitro antibacterial activity of Zn/Al-LDH-GA-loaded CMC films was assessed by disk diffusion and cell viability contact tests. The results showed the desired antibacterial activity against Staphylococcus aureus and Escherichia coli bacteria. Incorporating GA within CMC and CMC-Zn/Al-LDH films rereleased good cytocompatibility at the studied incubation time and different concentrations toward human normal HFF cell line than the free drug. The results of the present study indicated that the Zn/Al-LDH and Zn/Al-LDH-GA-loaded CMC have promising wound healing features to further develop a better future for clinical remedy of the different non-healing and hard-to-heal wounds.

Biogenic Selenium Nanoparticles Synthesized by L. brevis 23017 Enhance Aluminum Adjuvanticity and Make Up for its Disadvantage in Mice.

The most popular vaccine adjuvants are aluminum ones, which have significantly reduced the incidence and mortality of many diseases. However, aluminum-adjuvanted vaccines are constrained by their limited capacity to elicit cellular and mucosal immune responses, thus constraining their broader utilization. Biogenic selenium nanoparticles are a low-cost, environmentally friendly, low-toxicity, and highly bioactive form of selenium supplementation. Here, we purified selenium nanoparticles synthesized by Levilactobacillus brevis 23017 (L-SeNP) and characterized them using Fourier-transform infrared spectroscopy, energy-dispersive X-ray spectroscopy, scanning electron microscopy, and transmission electron microscopy. The results indicate that the L-SeNP has a particle size ranging from 30 to 200 nm and is coated with proteins and polysaccharides. Subsequently, we assessed the immune-enhancing properties of L-SeNP in combination with an adjuvant-inactivated Clostridium perfringens type A vaccine using a mouse model. The findings demonstrate that L-SeNP can elevate the IgG and SIgA titers in immunized mice and modulate the Th1/Th2 immune response, thereby enhancing the protective effect of aluminum-adjuvanted vaccines. Furthermore, we observed that L-SeNP increases selenoprotein expression and regulates oxidative stress in immunized mice, which may be how L-SeNP regulates immunity. In conclusion, L-SeNP has the potential to augment the immune response of aluminum adjuvant vaccines and compensate for their limitations in eliciting Th1 and mucosal immune responses.

Auxin acts upstream of nitric oxide to regulate cell wall xyloglucan and its aluminium-binding capacity in Arabidopsis thaliana.

MAIN CONCLUSION: Auxin acts upstream of NO through NOA and XXT5 pathways to regulate the binding capacity of the root cell wall to Al. In our previous study, we identified an unknown mechanism by which 1-naphthaleneacetic acid (NAA) decreased the fixation of aluminum (Al) in the cell wall. Here, we observed that external application of the nitric oxide (NO) donor S-nitrosoglutathion (GSNO) increased the inhibition of Al on root elongation. Further analysis indicated that GSNO could induce Al accumulation in the roots and root cell walls, which is consistent with lower xyloglucan content. In comparison to the Columbia-0 (Col-0) wild type (WT), endogenous NO-reduced mutants noa1 (NOA pathway) and nia1nia2 (NR pathway) were more resistant to Al, with lower root Al content, higher xyloglucan content, and more Al accumulation in the root cell walls. By contrast, the xxt5 mutant with reduced xyloglucan content exhibited an Al-sensitive phenotype. Interestingly, Al treatment increased the endogenous auxin and NO levels, and the auxin levels induced under Al stress further stimulated NO production. Auxin application reduced Al retention in hemicellulose and decreased the xyloglucan content, similar to the effects observed with GSNO. In yucca and aux1-7 mutants, exogenous application of NO resulted in responses similar to those of the WT, whereas exogenous auxin had little effect on the noa1 mutant under Al stress. In addition, as auxin had similar effects on the nia1nia2 mutant and the WT, exogenous auxin and NO had little effect on the xxt5 mutant under Al stress, further confirming that auxin acts upstream of NO through NOA and XXT5 pathways to regulate the binding capacity of the root cell wall to Al.

Effects of Chronic Aluminum Administration on Blood and Liver Iron-Related Parameters in Mice

In this study, the effects of chronically administered aluminum on iron metabolism-related parameters of liver and blood of mice were investigated. An additional purpose to determine how chronic aluminum administration together with vitamin E as an antioxidant to mice changed the parameters related to iron metabolism. For these purposes, we used 21 adult female Balb-c mice in this study. The animals were divided into three groups: one group with aluminum administered chronically, another group with aluminum plus vitamin E administered chronically, and the control group. Serum levels of hemoglobin, ferritin, iron, transferrin, hematocrit, total iron binding capacity (TIBC), as well as percentage of transferrin saturation were determined in all groups. In addition, the liver tissue levels of ferritin and iron were analyzed. Hemoglobin and hematocrit levels of the aluminum group and aluminum plus vitamin E group were significantly decreased compared to the control. In conclusion, no changes occurred in the serum iron related parameters although Al induced anemia in mice when Al administered chronically. There was an increase in the levels of liver iron and ferritin with Al, but Vit E had no effect on the changes of all blood and liver parameters caused by Al.

Lead ions but not other metallic ions increase resistance to hypotonic lysis in prenatal hemopoiesis red blood cells

Metals known to have toxic effects on exposed individuals (Aluminum (Al), Cadmium (Cd), Zinc (Zn) and lead (Pb)) were selected. Umbilical cord erythrocytes from normal newborns were incubated in isotonic media alone or with addition of Pb (20 microM), Cd, Zn or Al (concentration range: 20-250 microM). Red cells were then placed in media of diminishing tonicity, to measure cellular lysis and volume; the regression curves of percent lysis as a function of osmolarity were determined for each data set and the break points calculated. Resistance to lysis increased significantly in Pb treated cells whereas cells treated with the other metals did not differ from controls, even at concentrations ten times higher than that of Pb. Lead produced a reduction in cellular volume corrected by addition of quinidine (an inhibitor of potassium channels activation) to the cell suspension; on the other hand, quinidine did not modify the effect of lead on lysis sensitivity. These results suggest that the effect of lead on cell resistance to lysis might be mediated by changes in membrane structure. The other metals examined did not affect the variables studied.(AU)

Effects of aluminum sulfate on delta-aminolevulinate dehydratase from kidney, brain, and liver of adult mice

The purpose of the present study was to investigate the in vitro and in vivo effects of aluminum sulfate on delta-aminolevulinic acid dehydratase (ALA-D) activity from the brain, liver and kidney of adult mice (Swiss albine). In vitro experiments showed that the aluminum sulfate concentration needed to inhibit the enzyme activity was 1.0-5.0 mM (N = 3) in brain, 4.0-5.0 mM (N = 3) in liver and 0.0-5.0 mM (N = 3) in kidney. The in vivo experiments were performed on three groups for one month: 1) control animals (N = 8); 2) animals treated with 1 g per cent (34 mM) sodium citrate (N = 8) and 3) animals treated with 1 g per cent (34 mM) sodium citrate plus 3.3 g per cent (49.5 mM) aluminum sulfate (N = 8). Exposure to aluminum sulfate in drinking water inhibited ALA-D activity in kidney (23.3 + ou - 3.7 per cent, mean + ou - SEM, P<0.05 compared to control), but enhanced it in liver (31.2 + ou - 15.0 per cent, mean + ou - SEM, P<0.05). The concentrations of aluminum in the brain, liver and kidney of adult mice were determined by graphite furnace atomic absorption spectrometry. The aluminum concentrations increased significantly in the liver (527 + ou - 3.9 per cent, mean + ou - SEM, P<0.05) and kidney (283 + ou - 1.7 per cent, mean + ou - SEM, P<0.05) but did not change in the brain of aluminum-exposed mice. One of the most important and striking observations was the increase in hepatic aluminum concentration in the mice treated only with 1 g per cent sodium citrate (34 mM) (217 + ou - 1.5 per cent, mean + ou - SEM, P<0.05 compared to control). These results show that aluminum interferes with delta-aminolevulinate dehydratase activity in vitro and in vivo. The accumulation of this element was in the order: liver > kidney > brain. Furthermore, aluminum had only inhibitory properties in vitro, while in vivo it inhibited or stimulated the enzyme depending on the organ studied.

Aluminum induces lipid peroxidation and aggregation of human blood platelets

Aluminum (Al3+) intoxication is thought to play a major role in the development of Alzheimer's disease and in certain pathologic manifestations arising from long-term hemodialysis. Although the metal does not present redox capacity, it can stimulate tissue lipid peroxidation in animal models. Furthermore, in vitro studies have revealed that the fluoroaluminate complex induces diacyglycerol formation, 43-kDa protein phosphorylation and aggregation. Based on these observations, we postulated that Al3+-induced blood platelet aggregation was mediated by lipid peroxidation. Using chemiluminescence (CL) of luminol as an index of total lipid peroxidation capacity, we established a correlation between lipid peroxidation capacity and platelet aggregation. Al3+ (20-100 muM) stimulated CL production by human blood platelets as well as their aggregation. Incubation of the platelets with the antioxidants nor-dihydroguaiaretic acid (NDGA) (100 muM) and n-propyl gallate (NPG) (100 muM), inhibitors of the lipoxygenase pathway, completely prevented CL and platelet aggregation. Acetyl salicylic acid (ASA) (100 muM), an inhibitor of the cyclooxygenase pathway, was a weaker inhibitor of both events. These findings suggest that Al3+ stimulates lipid peroxidation and the lipoxygenase pathway in human blood platelets thereby causing their aggregation.

Efeito do aluminio em plaquetas humanas / Effect of aluminium in human platelets

O efeito do aluminio foi avaliado em plaquetas humanas de individuos clinicamente saudaveis atraves de estudos de agregacao, peroxidacao e fosforilacao de proteinas em residuo de tirosina com os inibidores plaquetarios "in vitro" (Trolox, AAS, NDGA, EDTA E DFO). Os resultados obtidos demonstraram que o aluminio foi capaz de promover peroxidacao lipidica com ativacao de proteina tirosina quinase (PTK) e agregacao plaquetaria. A ativacao plaquetaria pelo aluminio independe da formacao de metabolitos do acido araquidonico via ciclooxigenase. por outro lado, os eventos de agregacao e fosforilacao de PTK foram dependentes de peroxidacao enzimatica mediada preferencialmente por hidrolise de acido araquidonico, via ativacao de lipoxigenases

Inhibition of bovine brain acetylcholinesterase by aluminum

We report the in vitro inhibitory effect of very low concentrations of aluminum salts (IC50 = 4.1 x 10-12M) on bovine brain acetylcholinesterase (AChE). The enzymatic assays were performed using acetylcholine bromide in a buffered pH 7.4 solution at 37§C. The relevant enzyme interacting species is the Al3+ ion, whose concentrations were fixed at pM levels by a citrate metal ion buffer system. The IC50 demonstrates that Al3+ is a potent inhibitor of AChE