A simple hydroponic culture method for the development of a highly viable root system in Arabidopsis thaliana.
In the studies of nutritional absorption and metal toxicity in the root, it is important to grow plants without technical damage. We established a simple hydroponic culture system for Arabidopsis thaliana to obtain a healthy plant having a well-developed root system with many lateral roots. The phytotoxic effects of Cr, Cu, and Al ions were examined by FDA-PI staining using this culture system. The pattern of root inhibition varied with the ion, suggesting the usefulness of this culture system. (+info)
Binding of the transition state analog MgADP-fluoroaluminate to F1-ATPase.
Escherichia coli F1-ATPase from mutant betaY331W was potently inhibited by fluoroaluminate plus MgADP but not by MgADP alone. beta-Trp-331 fluorescence was used to measure MgADP binding to catalytic sites. Fluoroaluminate induced a very large increase in MgADP binding affinity at catalytic site one, a smaller increase at site two, and no effect at site three. Mutation of either of the critical catalytic site residues beta-Lys-155 or beta-Glu-181 to Gln abolished the effects of fluoroaluminate on MgADP binding. The results indicate that the MgADP-fluoroaluminate complex is a transition state analog and independently demonstrate that residues beta-Lys-155 and (particularly) beta-Glu-181 are important for generation and stabilization of the catalytic transition state. Dicyclohexylcarbodiimide-inhibited enzyme, with 1% residual steady-state ATPase, showed normal transition state formation as judged by fluoroaluminate-induced MgADP binding affinity changes, consistent with a proposed mechanism by which dicyclohexylcarbodiimide prevents a conformational interaction between catalytic sites but does not affect the catalytic step per se. The fluorescence technique should prove valuable for future transition state studies of F1-ATPase. (+info)
Fluorimetric determination of aluminum traces in hemodialysis solutions using Mordant Red 19.
A sensitive and accurate method for the spectrofluorimetric determination of trace levels of aluminum in hemodialysis solutions using Mordant Red 19 as the complexation reagent has been developed. The optimal experimental conditions for the concentration of fluorimetric reagent, pH, temperature, and the specific type of matrix are reported. The emission of the fluorescent metal chelate was measured at 555 nm, excitation at 478 nm. Linearity between emission intensity and aluminum concentration was found in the 2-20 ppb range in standard aluminum solutions. Limit of detection was 0.4 ppb. The aluminum amounts in some commercial hemodialysis solutions were determined by means of the extrapolation method. The proposed method proved to be suitable in terms of sensitivity and accuracy for the determination of aluminum in dialysis fluids. (+info)
Aluminum is a weak agonist for the calcium-sensing receptor.
BACKGROUND: Aluminum (Al3+) has diverse biological effects mediated through activation of a putative extracellular cation-sensing receptor. A recently identified calcium-sensing receptor (CaSR), which has been identified in target tissues for Al3+, may transduce some of the biological effects of Al3+. METHODS: To test this possibility, we transfected human embryonic kidney 293 (HEK 293) cells with a cDNA encoding the rat CaSR and evaluated CaSR expression by Western blot analysis and function by measurement of intracellular calcium ([Ca2+]i) levels and inositol monophosphate (IP1) generation following stimulation with Al3+ and a panel of CaSR agonists. RESULTS: The CaSR protein was detected by immunoblot analysis in cells transfected with the CaSR cDNA but not in nontransfected HEK 293 cells. In addition, [Ca2+]i levels and IP1 generation were enhanced in a dose-dependent fashion by additions of the CaSR agonists calcium (Ca2+), magnesium (Mg2+), gadolinium (Gd3+), and neomycin only in cells transfected with CaSR. To determine if Al3+ activated CaSR, we stimulated cells transfected with rat CaSR with 10 microM to 1 mM concentrations of Al3+. Concentrations of Al3+ in the range of 10 microM to 100 microM had no effect on [Ca2+]i levels or IP1 generation. In contrast, 1 mM Al3+ induced small but significant increases in both parameters. Whereas Gd3+ antagonized calcium-mediated activation of CaSR, pretreatment with Al3+ failed to block subsequent activation of rat CaSR by Ca2+, suggesting a distinct mechanism of Al3+ action. CONCLUSION: Al3+ is not a potent agonist for CaSR. Because Al3+ affects a variety of target tissues at micromolar concentrations, it seems unlikely that CaSR mediates these cellular actions of Al3+. (+info)
High-resolution PET imaging for in vivo monitoring of tumor response after photodynamic therapy in mice.
The aim of this study was to investigate the use of [18F]fluoro-2-deoxy-D-glucose (FDG) and a small-animal PET scanner to assess early tumor response after photodynamic therapy (PDT) in mice. PDT consists of intravenous administration of a photosensitizer that accumulates preferentially in tumor tissue, followed by local illumination of the tumor with red light. Two different photosensitizers were used: Photofrin (PII), which has been approved for clinical use, and disulfonated aluminum phthalocyanine (AIPcS), which is a second-generation drug. These drugs have been shown to induce tumor necrosis through different action mechanisms, i.e., mainly initial vascular stasis (PII) or direct tumor cell kill (AIPcS). FDG PET was used to follow both perfusion and metabolic activity in the tumor tissue. METHODS: The study was performed using a mouse model implanted with two contralateral murine mammary tumors (5 mm diameter x 2.5 mm thickness) on the back. Only one tumor was subjected to PDT, whereas the other tumor served as a control. A total of 13 mice were studied, 1 without illumination, 3 at 30 min and 3 at 2 h after PDT with both PII-PDT and AIPcS-PDT. Dynamic PET imaging of the mice, which were placed in pairs in a prostate position parallel to the transaxial planes of the Sherbrooke animal PET scanner, was performed after a bolus injection of 11 MBq (300 microCi) FDG. Blood samples were collected concurrently from 1 mouse during each study using an automated microvolumetric blood sampler. RESULTS: Analysis of the tumor time-activity curves showed that (a) scans during the first 3 min provided an estimate of tumor perfusion, as confirmed by the blood samples; (b) the tumor FDG uptake after 15 min was a direct measurement of tumor metabolism clearly demonstrating the relative efficacy of the two PDT drugs; and (c) the tumor tracer concentration in the interval 3-15 min after FDG injection is an appropriate indicator of the different mechanisms of tumor necrosis through indirect vascular stasis (PII) or direct cell kill (AIPcS). CONCLUSION: This pilot study confirmed the feasibility of using dynamic in vivo PET imaging for assessing early tumor response to PDT in mice. (+info)
Evidence that cytosolic phospholipase A2 is down-regulated by protein kinase C in intact human platelets stimulated with fluoroaluminate.
We reported that protein kinase C (PKC) inhibitors increase the release of arachidonic acid induced by fluoroaluminate (AlF4-), an unspecific G-protein activator, in intact human platelets. Now we demonstrate that this effect is independent of the extracellular Ca2+ concentration and that AlF4(-)-induced release of AA is abolished by BAPTA, an intracellular Ca2+ chelator, even in the presence of GF 109203X, a specific and potent PKC inhibitor. This compound also blocks the liberation of the secretory phospholipase A2 in the extracellular medium, indicating that this enzyme is not involved in the potentiation of arachidonic acid by PKC inhibitors. On the other hand, the latter effect is completely abolished by treatment of platelets with AACOCF3, a specific inhibitor of cytosolic phospholipase A2 (cPLA2). These observations indicate that cPLA2 is responsible for the AlF4(-)-induced release of arachidonic acid by a mechanism that is down-regulated by PKC. (+info)
A preliminary study of dietary aluminium intake and risk of Alzheimer's disease.
BACKGROUND: epidemiological studies of Alzheimer's disease and aluminium intake have focused on aluminium in drinking water. There have been no studies investigating the relation between the disease and the consumption of foods containing large amounts of aluminium additives. OBJECTIVES: to conduct a pilot study to determine whether dietary intake of aluminium additives differs in individuals with and without Alzheimer's disease. DESIGN: matched case-control study. Controls were matched to cases on age, gender and date of admission to the centre. SETTING: Syracuse, New York, USA. SUBJECTS: 46 participants comprising 23 matched sets. METHODS: residents of the Loretto Geriatric Center with and without newly-diagnosed Alzheimer's disease were selected. Next-of-kin were asked to complete information on the resident's medical history, lifestyle behaviour and dietary intake before admission to the centre. An expanded form of the Health Habits and History Questionnaire was used to determine dietary intake. Consumption of foods containing elevated levels of aluminium additives was compared between cases and controls. RESULTS: the crude odds ratio for daily intake of foods containing high levels of aluminium was 2.0 and, when adjusted for covariates, was 8.6 (P=0.19). Intake of pancakes, waffles, biscuits, muffins, cornbread and/or corn tortillas differed significantly (P=0.025) between cases and controls. Adjusted odds ratios were also elevated for grain product desserts, American cheese, chocolate pudding or beverages, salt and chewing gum. However, the odds ratio was not elevated for tea consumption. CONCLUSION: past consumption of foods containing large amounts of aluminium additives differed between people with Alzheimer's disease and controls, suggesting that dietary intake of aluminium may affect the risk of developing this disease. Larger studies are warranted to corroborate or refute these preliminary findings. (+info)
Effects of aluminum potassium sulfate on learning, memory, and cholinergic system in mice.
AIM: To study the relationship between aluminum potassium sulfate (APS) and memory deficits of mice. METHODS: 30, 60, or 90 d after the mice were given daily APS i.g., the step-through latency (STL) was determined with a passive avoidance task. Aluminum (Al) contents in brain and blood were assayed with atomic absorption spectrophotometry. Acetylcholine (ACh) content in brain was determined with chemiluminescent method and choline acetyltransferase (ChAT) activity was measured radiochemically. RESULTS: APS 1 g.kg-1 increased blood-Al only after 30 d. After 60 d, STL, ACh content and ChAT activity decreased by 46.4%, 8.5%, and 22.6%, respectively. These parameters decreased by 50%, 11.1%, and 27.8%, respectively, with increased Al in blood and brain, after 90 d. APS 0.25 g.kg-1 had no effects on mice except blood-Al. In ethylcholine mustard aziridium chloride (AF64A) treated mice, APS 1 g.kg-1 only increased blood and brain-Al. CONCLUSION: The intake of APS 1 g.kg-1.d-1 for 60 d induced learning and memory deficits in mice. (+info)