EDTA Therapy for Vascular Disease

Chelation Natural Miracle For Protecting Your Heart

Chelation therapy has been conclusively shown to be up to 82 Percent Effective at dissolving the plaque that blocks arteries! In the ebook Chelation Natural Miracle For Protecting Your Heart you'll discover: The frustrating reason many doctors are ignoring Edta chelationor even openly rejecting it. The deadly heart surgeries even the American Heart Association admits are unnecessary. The hidden signs and symptoms of heart attacks and strokes? Are you in danger right now?. The average number of years stripped away by heart and vessel disease. Can you get them back?. The newest set of risk factors for heart disease (they'll likely surprise you!). Shady government practices that protect Big Pharma and keep Edta chelation out of the public eye. How the Roman Empire could have been savedif only they'd known about Edta chelation. Why Edta chelation is guaranteed to be safeeven in extremely high doses. (It puts aspirin to shame!). The shocking truth about plaque in young childrenand how to keep your little ones safer. Why dentists, artists and welders need Edta chelationand whether your workplace is dangerous too. The differences between IV and oral chelationand which kind of Edta is right for you. Other forms of chelationand how these little-known treatments can dramatically boost your health.

Chelation Natural Miracle For Protecting Your Heart Summary


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Author: Michael Cutler, M.D.
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Iron Chelation Therapy

Iron chelation therapy inhibits erythrocytic Plasmodium parasites in vitro and in vivo (Mabeza et al. 1999). Ultrastructural studies show a stage specific effect on a trophozoite-to-schizont stage transition with an enlarged nucleus rather than an enlarged digestive vacuole as seen with the quinolines (Atkinson et al. 1991). Iron chelation also inhibits mouse malaria development in hepatocytes at appropriate concentrations (Loyevsky et al. 1999b). The mechanism of action with diverse groups of iron chelators can be grouped to either withholding of iron or by formation of a toxic complex with iron (Mabeza et al. 1999). Iron withholding has a direct effect on the ribonucleotide reductase necessary for DNA synthesis and or 6-aminolevulinate synthase for heme synthesis. Because of minimal effects on host cells and concurrent development of iron chelators as adjunctive cancer chemotherapy, several trials of iron chelators have been performed for both severe malaria and uncompli cated...

Endocannabinoid Uptake Studies In Vitro Assay of Endocannabinoid Uptake

COS cells (107) grown to confluence in flasks in DMEM (Life Technologies, Gaithersburg, MD) containing 10 FCS (Life Technologies) were split 1 2, harvested the next day using trypsin EDTA, centrifuged (200g) for 10 min at 4 C, washed with sterile HEBS (20 mM HEPES, 137 mM NaCl, 5 mM KCl, 0.7 mM Na2HPO4, and 6 mM dextrose), recentrifuged, and resuspended at 107 cells mL in 4 C HEBS. Then, 0.9 mL of suspended cells was transfectedby electroporation at 300 V and 1100 F in 400-mm Gene Pulser cuvettes (Bio-Rad) containing 20 g of plasmid DNA and 500 g of fish sperm DNA (Boehringer-Mannheim, Mannheim, Germany) using a geneZAPPER 450 2500 (IBI, New Haven, CT).

General Information

The use of calcitetracemate in the treatment of lead poisoning has been described (1). Between 1993 and 2000, 45 adult patients consulted the Poison Centre of Marseilles, France after lead exposure (9 women, 36 men, average age 44, range 22-76 years). In 22 patients, a calcitetracemate provocation test was negative. Six patients with a positive test refused to be treated the other 16 were treated with a total of 58 courses of calcitetracemate by infusion 500 mg bd for 5 days. The mean blood lead concentration in these 16 patients was 566, range 320-943 ng ml the mean urinary lead excretion was 3011, range 789-7229 mg day. The mean amount of lead eliminated in the urine during chelation therapy was 30 912 mg. In 12 patients in whom lead exposure ended after the diagnosis of lead poisoning, chelation therapy reduced the blood lead concentration by 69 . In four patients in whom exposure continued during treatment, the blood lead concentration fell by only 7 . In the 16 treated patients,...

Susceptibility Factors Age

Overload has been established and is associated with a significant reduction in mean body length (151,152). Impairment of growth is associated with a rickets-like syndrome and joint stiffness. Metabolic studies have shown reduced hair and leukocyte zinc concentrations and leukocyte alkaline phosphatase activity. Retardation of growth may be related to chelation of trace elements (for example zinc), to a direct toxic effect of unchelated deferoxamine by inhibiting critical iron-dependent enzymes, or to both. It is advised that in thalassemia major, treatment with deferoxamine be started only after iron accumulation is established, that is at around 3 years of age, after 20-30 blood transfusions, when ferritin concentrations are in the range 800-1000 ng ml. Deferoxamine doses should be established on the basis of studies of iron balance and dose-response curves, and longitudinal growth monitoring is warranted.

Other features of the patient

A possible relation has been suggested between toxicity and the ratio of metabolite B of deferoxamine to un-metabolized deferoxamine (SEDA-18, 250) and there is a relation between regular deferoxamine treatment in respect to the degree of iron overload, as defined by the therapeutic index, that is the daily dose of deferox-amine in mg kg day divided by the serum ferritin concentration in ng ml (SED-13, 621), and the ratio of metabolite B to total deferoxamine (deferoxamine plus ferrioxamine) in the plasma or urine (153). This is consistent with the hypothesis that metabolite B of deferox-amine, which is a product of the intercellular metabolism of iron-free but not iron-bound deferoxamine, inversely reflects the availability of iron in the plasma compartment. In patients who receive a high amount of chelation (measured as mean daily dose of deferoxamine in mg kg) in relation to iron stores (as reflected by serum ferritin in ng ml), the proportion of iron-free deferoxamine that is...

Calcineurin inhibitors

Oral absorption of Sandimmune is low (5-30 ) and highly variable, ranging from 4 to 89 in renal and liver transplant patients (33,34). Absorption of the microemulsion formulation is more consistent, averaging approximately 40 (35). Peak blood concentrations typically occur between 1-3 and 2-6 h following oral administration of Neoral and Sandimmune, respectively (33,36,37). Absorption can be delayed for several hours in a subgroup of patients. Because CsA is lipophilic, it crosses most biologic membranes and has a wide tissue distribution (38). CsA is highly bound to plasma proteins ( 90 to lipoproteins), with the majority of CsA localizing in erythrocytes. The distribution of CsA between plasma and erythrocytes is temperature-dependent and varies with changes in hematocrit (39). Because of the potential for artifactural redistribution of CsA during specimen processing because of ambient temperature fluctuations, ethylenediaminetetraacetic acid (EDTA)-anticoagulated whole blood should...

Preanalytic Variables

Plasma or serum can be used to measure MPA and free MPA blood concentrations (187). However, plasma from EDTA-anticoagulated whole blood is the recommended specimen of choice because the same sample can be used to measure whole blood CsA, tacrolimus, and sirolimus (196). MPA and MPAG are stable in whole blood and plasma samples at room temperature for at least 4h (197). Plasma samples are stable at 4 C for 4 days and at least 11 months when stored at 20 C (196). Free MPA is stable for at least 6 months when stored at 20 C (198). Thawing and re-freezing of plasma samples can be performed up to four times without significant loss of MPA (199). When monitoring MPA during intravenous infusion of mycophenolate mofetil, whole blood samples should be immediately placed in ice and the plasma separated within 30min (200,201). This is because mycophenolate mofetil is very unstable and rapidly undergoes temperature-dependent degradation to MPA in whole blood samples placed at room temperature...

Modulation of the Release of Endocannabinoids by Other Neurotransmitters

The essential finding that postsynaptic increases in intracellular Ca2+ can induce endocannabinoid release has also led many investigators to an important corollary hypothesis that activation of postsynaptic receptors known to be coupled to the mobilization of intracellular Ca2+ should also promote endocannabinoid release. At both GABAergic and glutamatergic synapses in the CNS, a number of G-protein-coupled receptor systems are known to be linked to Ca2+ mobilization, including metabotropic glutamate receptors (mGluRs Anwyl, 1999 De Blasi et al., 2001) and muscarinic cholinergic receptors (Kerlavage et al., 1987). Thus, several recent studies have investigated whether activation of these receptor systems can also facilitate endocannabinoid release. In hippocampal slices, Varma et al. (2001) showed that activation of group I mGluRs (mGluRl and or mGluR5) reduced GABA release onto CA1 pyramidal cells and that this effect was blocked by a CBj receptor antagonist (Varma et al., 2001)....

LTD at Glutamatergic Synapses in the Dorsal Striatum

The basal ganglia contain some of the highest densities of CBj receptors in the CNS (Herkenham et al., 1991 Herkenham, 1992). As was the case for other brain regions, pharmacological and electrophysiological techniques confirmed the presynaptic inhibitory effects of CBj receptors at both glutamatergic and GABAergic terminals within various basal ganglia nuclei (see Table 9.1). However, as was the case in the hippocampus and cerebellum, the fundamental link between the presence of these functional CB1 receptors and their endogenous ligands remained to be established. Anandamide release in the dorsolateral striatum, and its enhancement by dopamine D2 receptor agonists, was first demonstrated using microdialysis techniques (Giuffrida et al., 1999). Such an approach, although well-suited to addressing global or widespread activation of endocannabinoid systems, cannot assess endocannabinoid release and activity at the synaptic level. Thus, whole-cell and extracellular recording techniques...

Preparation of Cell Membrane Homogenates

The membrane homogenate preparation procedure isolates cell membranes and membrane-associated macromolecules and assures their homogeneity for assaying small fractions of the preparation in each of many test tubes. It removes the majority of soluble macromolecules, small molecules (including endogenous receptor ligands), and minerals via multiple centrifugation steps each followed by resuspension in fresh buffer containing the divalent ion chelator ethylenediamine tetraacetic acid (EDTA). Some prefer a P2 preparation, which involves discarding an initial pellet generated by a low-speed centrifugation step, but this appears to be unnecessary for these assays.

Treatment Of Toxic Element Poisoning

Removal of the poisoned individual from the environmental source (or vice versa) responsible for the toxic metal contamination and supportive care are generally all that is required for decontamination. In some cases, however, chelation therapy using organic compounds to bind and remove metals from the body may be warranted. Because chelation therapy may pose life-threatening side effects, due to the simultaneous chelation of essential elements such as calcium, care must be taken to manage the administration of chelation therapy (53-56). In any case, monitoring the elimination of toxic element exposure and confirmation of effective decontamination are useful when the appropriate laboratory specimen is utilized. For lead and iron, blood is a good specimen for monitoring decontamination, whereas urine is preferred for monitoring decontamination of mercury and most other toxic elements. Chelation therapy to treat children with blood lead concentrations of 20-44 g dL showed no benefit...

Preparation of Protein Extracts

Sonication buffer for quick and easy protocol (100 mL) 2 mL of 1 M N-(2-hydroxyethyl) piperazine-N'-(2-ethanesulfonic acid) (HEPES)-KOH buffer, pH 7.9 (20 mM), 25 mL of 100 glycerol (25 ), 25 mL of 2 M KCl (0.5 M), 150 L of 1 M MgCl2 (1.5 mM), and 40 L of 0.5 M sodium EDTA, pH 8.0 (0.2 mM). Bring to 100 mL with distilled water. Additives Before use add to an aliquot of sonication buffer (e.g., 1 mL) 1 L of 1 MDTT mL (1 mM) and 1X mammalian protease inhibitor cocktail (commercially available from Calbiochem cat. no. 539134) (see Note 1). If state of phosphorylation is important, add 1 L of 1 M NaF mL (1 mM), 1 L of 5 Mokadaic acid mL (5 nM), and 1 L of 100 mM Na3VO4 mL (100 M). 2. Buffer A for crude nuclear extract (100 mL) 1 mL of 1 MHEPES-KOH, pH 7.9 (10 mM), 150 L of 1 M MgCl2 (1.5 mM), 500 L of 2 M KCl (10 mM), 200 L of 0.5 M sodium EDTA, pH 8.0 (1 mM), and 25 mL of 100 glycerol (25 ). Bring to 100 mL with distilled water. Additives Before use add to an aliquot 1 L of 1 M DTT mL (1...

Photographicprinting chemicals

Many of these substances arc highly irritant and corrosive (see overview in Schardein 2000, Gilstrap 1998, Paul 1993). The most commonly used chemicals are acetic acid, ammonium sulfate, ammonium thiocyanate, ammonium thiosulfate, bromine potassium bromide, citric acid, diethylenetriaminepenta acetic acid (DTPA), ethylenediamine tetra acetic acid (EDTA ferric ammonium salt of EDTA), glycol ethers, hydrocarbon solvents, hydroxy-lamine, 3-phenylenediamine, 4 -phenylenediamine, potassium carbonate, sodium benzoate, sodium sulfite, and sulfur dioxide.

Phenoxyacetic acid derivatives and chlorinated dibenzodioxins

In general, maternal blood lead concentrations within the normal range (i.e. 5 ng dl. In adults, the blood lead concentrations regarded as toxic and requiring chelation therapy are in the range of 60pg dl. In pregnant women, blood lead concentrations 30 tg dl would be cause for concern. At concentrations which cause severe maternal toxicity ( 100 .ig dl), an increased risk of fetal loss may occur. In such circumstances, the uterine muscles relax and the fetus is expelled from the uterus. It is not known whether this is due entirely to the high concentrations of lead per se, or whether it is secondary to the maternal toxicity (see overview in Schardein 2000, Rabinowitz 1988, Barlow 1982, Scanlon 1975). In children with brain dysfunction following in utero exposure to lead, the blood concentrations of lead are usually 35 pg dl. Lead concentrations of 40 tg dl in children are regarded as toxic, and require chelation therapy. If indicated, dental amalgam fillings may be restored during...

Antioxidant Scavanger Action

In our own laboratory, we assessed the antioxidant activity of EGb761 in in vitro experiments. The formation of TBA reactive substances (TBARS) was used as an index of lipoperoxidation (LPO) in all forebrain membranes 34 . The degree of peroxidation (lipoperoxidation index LPO) increased gradually with increasing time (30 to 60 min) of incubation in the medium containing 0.1 mmol l FeSO4 EDTA plus 1 mmol l H2O2 mg of protein. The addition of 50 or 100 J.l ml EGb761 significantly protects the generation of LPO products in cerebral membranes. In addition, oxidative modifications in protein measured by the content of total sulfhydryl group level was protected up to 75 to 81 by the addition of 50 or 100 J.l ml EGb.

Other Therapies 1151 Razoxane

Razoxane is a cyclized analog of ethylenediaminetetraacetic acid (EDTA) that was first synthesized in the late 1960s (see Structure 11.5). It has limited activity in the leukemias and is now little used in the clinic. Not surprisingly, line EDTA, razoxane possesses intracellular iron-chelating activity.

Setting Up the RNA Workplace

RNA is very susceptible to hydrolysis by RNases. RNases are present not only in tissues, but on the skin of investigators, and can thus be reintroduced into the preparation at any time. RNases are very difficult to inactivate. They are resistant to metal chelating agents, are active over a wide pH range, and can survive prolonged boiling or autoclaving. Indeed, if autoclaved reagents contain microorganisms, their RNases can be set free during autoclaving. Fortunately, RNases rely on histidine residues for catalytic activity and they can be inactivated by the alkylating agent DEPC, by 3 hydrogen peroxide solutions, 1 SDS solutions, or 1 N sodium hydroxide solutions. Glassware can also be baked at 180 C for several hours. The following steps should be taken before starting to work with RNA

Common Household Chemicals as Urinary Adulterants

Schwarzhoff and Cody studied the effect of 16 different adulterating agents ammonia-based cleaner, L-ascorbic acid, Visine eye drops, Drano, Golden Seal root, lemon juice, lime solvent, Clorox, liquid hand soap, methanol, sodium chloride, tribasic potassium phosphate, toilet bowl cleaner (Vanish, Drackett Products), white vinegar, ionic detergent (Multi-Terge) and whole blood anticoagulated with EDTA on FPIA analysis of urine for abused drugs. The authors tested these adulterating agents at 10 by volume concentration of urine with the exception of Golden Seal because of the insolubility. For Golden Seal tea, one capsule was suspended in 60 mL urine. Out of six drugs tested (cocaine metabolites, amphetamines, opiates, phencyclidine, cannabinoid and barbiturates), the cannabinoid test was most susceptible to adulteration. Approximately half of the agents (ascorbic acid, vinegar, bleach, lime solvent, Visine eye drops and Golden Seal) tested caused false negatives. Both cannabinoid and...

Adverse Reactions and Toxicity

Toxicity of a contrast medium is attributed to its molecular structure, solution property, formulation, and the amount used as a dose. Toxicity may be further classified as chemotoxic-ity, osmotoxicity, and ion toxicity (407, 459). Chemotoxicity is related to a molecular structure that allows binding to proteins, leading to interaction with biomacromolecules such as enzymes, cell membranes, and cell components. Osmotoxicity is attributed to the hyper-osmolality of contrast media, and a marked difference exists between the osmolalities of ionic and nonionic contrast media. Ion toxicity refers to adverse reactions as a consequence of too high or too low ion concentrations in contrast media that interfere with cellular function. Contrast media formulations containing citrate and EDTA buffers can interfere with serum Ca2+ and affect myocardial contractility function. Contrast media are transported by the cardiovascular system and are excreted by the kidneys. Any reactions to contrast...

Plasmodium Iron Sources and Pathways

The Plasmodium parasite requires iron for DNA synthesis, glycolysis, pyrimi-dine synthesis, heme synthesis and electron transport. Debate continues on the critical source of iron for the intraerythrocytic parasite. An important paradox regarding iron is that millimolar desferrioxamine (DFO), an iron chelator, is cytostatic for mammalian cells and bacteria, while a 60-fold lower concentration of 15 M DFO is cytocidal for P. falciparum despite the availability of 20 mM heme iron in hemoglobin (Mabeza et al. 1999). Postulated available sources susceptible to chelation include (1) extracellular iron from transferrin or free iron in the media (2) intracellular iron bound to low molecular weight proteins, heme iron or erythrocyte ferritin iron. Lovesky has identified an intraerythrocytic labile pool of bioavailable iron with the fluorophore calcein (Loyevsky et al. 1999a). In the infected erythrocyte most of the labile pool of iron is in the parasite. In addition, uninfected erythrocytes...

Targeting Hiv Rna with Small Molecules

Far from being a passive carrier of genetic code, RNA is intimately involved in a wide range of biological processes including chemical catalysis and information storage. The vast majority of cellular RNAs form higher order structures with other cellular components to facilitate the exchange of biochemical information. Successful molecular recognition of RNA often precedes catalytic events that are essential to a wide range of cellular activities, including initiation of DNA replication 3 , extension of the telomeric regions of chromosomes 4 , splicing of pre-mRNA 5 , and iron chelation 6 . In addition, RNA serves as the primary genome of most pathogenic viruses 7J.

Organs and Systems Nervous system

There have been reports of taste disturbance in patients taking ACE inhibitors, tentatively attributed to chelation of metal ions. However, no mechanism for taste loss due to angiotensin-II receptor blockers has been proposed. Reversible loss of taste discrimination has been reported with losartan (6). In two cases dysgeusia occurred some time after switching from an ACE inhibitor to losartan (1 week in one case and 3 months in the other). In both cases the dysgeusia disappeared after withdrawal of losartan (timing not reported in one case, 1 week after in the other) (7). The authors referred to previously reported cases, one with losartan (8) and one with valsartan (9), and to a personal communication from the manufacturer of 12 other cases within a large safety monitoring program.

Therapeutic Drug Monitoring of Immunosuppressant Drugs

Whole blood (EDTA) Whole blood (EDTA) Whole blood (EDTA) Whole blood (EDTA) EDTA, ethylenediaminetetraacetic acid. a , 75 , 100 , 100 , 150 The costs are based on published charge for these tests in our hospital laboratory and reference laboratories. EDTA, ethylenediaminetetraacetic acid. a , 75 , 100 , 100 , 150 The costs are based on published charge for these tests in our hospital laboratory and reference laboratories.

Triplex Cleaving Agents

Addition of an EDTA moiety to a triplex-binding ligand can convert it into a triplex-specific cleavage agent in the presence of Fe2+ and a reducing agent. EDTA conjugates, attached to the alkyamino side chain, have been prepared for BePI, BgPI 26 and BQQ 130 . The ligands cleave DNA at sites to which a triplex-forming oligonucleotide is bound, though the background cleavage in non-targeted duplex DNA is greater than that produced by EDTA-tethered oligonucleotides 3 . BgPI and BQQ conjugates produce double-stranded cuts at triplex target sites, cleaving the purine and pyrimidine strands of the duplex with equal efficiency. As a result of their selectivity for triplex DNA these agents can be used to cleave plasmid DNA at a single site to which a triplex-forming oligonucleotide is bound. In contrast BePI-conjugates only cleave the duplex pyrimidine strand while the purine strand is actually protected from background cutting. As a result, this conjugate cannot be used to produce...

Oxidation chemistry of 5ht and da

A widely used HO-generating system consists of AA, traces of Fe2+-ethylenediaminetetraacetic acid (EDTA), H2O2, and molecular oxygen (Udenfriend et al. 1954). In this medium, HO - is formed by decomposition of H2O2 by the Fenton reaction (Walling 1975) and the resulting Fe3+ is reduced back to Fe2+ by AA (equation 1). Thus, as long as AA and H2O2 are available, HO- is formed by this cyclic reaction. When incubated with this HO-generating system, 5-HT is oxidized extremely rapidly (Wrona et al. 1995). The initial step in the reaction involves HO- attack on 5-HT to give the 4,5-(1), 2,5-(2) (1) and 5,6-(3)-dihydroxytryptamine radicals, which are then oxidized by a second molecule of HO- to give 2,5-DHT, 4,5-DHT, and 5,6-DHT in approximate yields of 60 percent, 24 percent, and 11 percent, respectively (figure 1). However, 4,5-DHT is not an isolatable product owing to its very facile oxidation by molecular oxygen to give tryptamine-4,5-dione (T-4,5-D) in a reaction that forms H2O2 as a...

Laboratory Analysis Of Tcas

It is important to stress the need to avoid serum or plasma separator tubes when collecting samples for analysis of TCAs. Nyberg and Martensson (18) studied several types of blood collection tubes for stability of amitriptyline, imipramine, clomipramine, and their mono-demethylated metabolites collected in these tubes. Ethylenediaminete-traacetic acid (EDTA) tubes were most suitable and serum separator gel tubes were unsuitable because of loss of more than 40 of drug concentrations on storage. The losses were not caused by redistribution between blood cells and plasma but occurred mainly because of sample contact with serum or plasma separator gel or the caps of the tubes. Dasgupta et al. (19) studied Greiner serum separator gel tubes for stability of TCAs and many other drugs, and concluded that these tubes are not suitable for blood collection for analysis of TCAs.

Cisplatin carboplatin and oxaliplatin

However, it is still debatable which method most accurately predicts individual values of GFR or creatinine clearance. Whereas the Cr-EDTA method is the most accurate method of estimating GFR, most clinicians do not use it routinely, preferring to collect urine for estimation of creatinine clearance. Alternatively, the use ofspecial formulae has been recommended. A new formula (Wright's formula) may provide a more accurate estimate of GFR than the formulae of Cockcroft & Gault or Jelliffe (27C, 28C, 29c). Furthermore, the formulae used for conventional carboplatin chemotherapy cannot translate into high-dose chemotherapy in terms of prediction of myelosuppression or of response rates, because there is a poor correlation between the calculated and the real serum concentrations ofcarboplatin.


Inhibition of mature osteoclast function, promotion of osteoclast apoptosis, and interference with osteoblast-mediated osteoclast activation. In patients with osteoporosis, pamidronate increased bone mineral density by 6.8 over 2.2 years (11). The frequently observed adverse effects with pamidronate therapy were gastrointestinal gastritis due to either a local reaction when a high concentration of the drug stays in the mucus or by chelation with calcium ions (12).

Specimen Types

Serum and plasma are the most common type of specimen used in TDM. These specimens may be interfered by all four types of interferents covered in this chapter. Additionally, collection tube additives for plasma or whole blood specimens, such as ethylene-diamino-tetra-acetate (EDTA), heparin, citrate, fluoride, and oxalate may chelate metal ions and thus interfere with label enzymes such as alkaline phosphatase and thus generating false-positive or false-negative results.


Several different mechanisms have been suggested to explain the biological activity of the anthracyclines, and controversy still exists about the relative importance of each. The first centers on the fact that the planar ring system inserts between two DNA base pairs perpendicular to the long axis of the double helix, with the amino sugar conferring stability on the adduct through hydrogen bonding interactions with the sugar phosphate backbone. It is known from in vitro experiments that the adduct formed is sufficiently stable to interfere with DNA processing, including transcription. Second, the anthracyclines are known to form complexes (i.e., ternary complexes) with topoisomerase enzymes and DNA, which can lead to strand breaks. Third, binding to cell membranes has been observed, and this may alter membrane fluidity and ion transport as well as disturb various biochemical equilibria in the cell. Lastly, generation of semiquinone species can lead to free radical or hydroxy radical...


10 mM Tris-HCl-1 mM EDTA buffer (TE buffer) 1 mL of 1 M Tris-HCl, pH 7.5, and 200 L of 0.5 MEDTA, pH 8.0, in 99 mL of dH2O. 8. Hybridization buffer 23.8 mL of formamide 0.95 mL of 1 M Tris-HCl, pH 7.4, 0.19 mL of 250 mM EDTA, pH 8.0, 3.75 mL of 4 M NaCl, 9.52 mL of 50 (w v) dextran sulfate, 0.95 mL of 50X Denhardt's solution. Add dH2O to 40 mL and store indefinitely at -20 C.

Ascorbic acid

There is a complex interaction between ascorbic acid, iron, and deferoxamine (157). Ascorbic acid appears to be essential for the mobilization of stored iron into a labile pool, available for chelation therapy. As a result, the administration of deferoxamine to patients with ascorbic acid deficiency will have a limited effect. On the other hand, ascorbic acid increases the toxic effects of iron, especially on the heart. In order to prevent serious cardiac dysrhyth-mias, ascorbic acid should only be given after deferoxamine infusion has been started and adequate serum concentrations of deferoxamine have been reached (SEDA-8, 239) (158). Ascorbic acid reduces ferric to ferrous iron, and it is the reduced iron that is responsible for the generation of highly reactive free radicals, causing tissue injury (157). Ascorbic acid up to 2 g day stimulates deferoxamine-mediated urinary iron excretion, probably without adverse cardiac results (159). However, a report of the precipitation of...


Reduced anticoagulation has been reported in a patient taking warfarin when he was given chelation therapy (25). A 64-year-old man with coronary artery disease taking atenolol, lisinopril, atorvastatin, glyceryl trinitrate, triamterene, hydrochlorothiazide, omeprazole, nizatidine, enteric-coated aspirin, and a liquid multivitamin formulation containing spinach and broccoli extracts was given warfarin for bilateral pulmonary embolism. About 3 weeks later he received chelation therapy with a cocktail including sodium edetate 3 g and heparin 2500 units. The next day his INR had fallen from 2.6 to 1.6. He denied having missed a dose of warfarin, having taken other drugs, or having made dietary changes. The dose of warfarin was increased and the INR reached the target range.


EDTA-anticoagulated whole blood is the recommended specimen matrix (132). This is because almost all of the sirolimus ( 95 ) is concentrated in erythrocytes, and plasma levels are too low for most analytical methods (134). Whole blood samples are stable for 10 days at ambient temperature (141), at least 1 week at 30-34 C (141, 142), 30 days at 4 C (143), and at least 2 months at -40 C (143). Whole blood samples can withstand three freeze-thaw cycles without altering measured sirolimus concentrations (141,142).

Alzheimers Disease

Accumulation of P-amyloid is associated with Alzheimer's disease development its toxicity in cultures of hippocampal neurons is mediated via ROS, lipid peroxidation, activation of the caspase cascade, and apoptosis. When such cells were co-exposed to EGCG, incidence of the latter three events decreased in a manner independent of p53, Bax, Bcl-xL, and cyclooxygenase (COX) 7 . The cytotoxicity of amyloid proteins appears to rely largely upon the formation of well-ordered fibrillar assemblies. Polyphenols inhibit this formation independently of their antioxidant activity 8 . Iron chelation by EGCG also reduces the aggregation of a major component of neurofibrillary tangles, hyperphosphorylated tau, in the brains of these patients 9 . ROS generation is partially dependent upon processes requiring intracellular iron, including the Fenton reaction. Dysregulation of cellular iron homeostasis, including uptake, distribution, transport, and storage, is a causal factor in the patho-genesis of...


Elevated toxic element concentrations in a specimen should always be confirmed with secondary testing to ensure that the elevated result was not due to external contamination. Sources of external contamination include collection materials, the specimen donor, the environment, transport containers, or assay reagents. As a result, certified trace element-free blood collection containers are available. Royal-blue top blood collection tubes, available with or without anticoagulant, are certified as trace element-free for most clinically important elements. Tan top tubes ethylene diamine tetra acetate (EDTA) anticoagulant are certified as lead-free, so can, in addition to the royal blue tubes, be used for lead testing. Laboratories that perform trace element testing are often physically isolated from other laboratory areas, and pay particular attention to sources of external contamination such as labware materials, dust control, and airflow dynamics. Certified trace metal-free reagents are...


Chelation therapy with intravenous desferoxamine is indicated if the serum iron level is 55 n mol l or if an overdose was clearly taken and the pregnant woman develops seizures, is unconscious, or presents with circulatory shock. In these cases, there is not time to wait for serum iron levels.


Carbomers are acrylic acid polymers that are used for a variety of purposes. They are used in pharmaceutical processes as suspending agents, gel bases, emulsifiers, and binding agents. They are also used as artificial tears. Carbomer 974P is a carbomer that is the major non-aqueous component (5 polymer, 94 water) of a proprietary formulation called BufferGel, which also contains dibasic potassium phosphate, magnesium sulfate, dibasic sodium phosphate, sorbic acid, monobasic sodium phosphate, and disodium EDTA. It is a buffering agent, a microbicide, and a spermicide. It is formulated in an aqueous gel at pH 3.9, which acidifies twice its volume of semen to a pH of 5 and maintains the protective acidity of the vagina. It is used vaginally as a spermicide that also protects against HIV infection and possibly other sexually transmitted diseases.

Detecting Monoamines

Twenty-minute samples were injected, without any purification, in a HPLC equipped with a reversed-phase ODS column (LC 18 DB Supelcosil, Supelco, Bellefonte, PA) and an electrochemical detector (BAS, Lafayette, IN) to quantitate DA, 3,4-dihydroxyphenylacetic acid (DOPAC), and homovanillic acid (HVA). The mobile phase was 0.23 M sodium acetate, 0.015 M citric acid, 100 mg L EDTA pH 5.5. The electrode used was a silicon carbon paste TL-3 working electrode with potential set at +0.65 V in oxidation, coupled with an Ag AgCl reference electrode. Experiments were always performed 24 h after probe implantation. Perfusion Ringer (147.2 mM NaCl, 3.4 mM Ca Cl2, and 4 mM KCl) flow was 2 L min (11). Dialysate samples (10 or 20 L) were injected without purification into an HPLC equipped with a reversed-phase column (LC-18 DB, 15 cm, 5 m particle size Supelco) and a coulometric detector with a 5014 A cell (ESA Coulochem II, Bedford, MA) to quantitate DA and NA. The first electrode of the detector...

Iron Fe

Fe is unavailable to plants when the pH of the water or soil is too high. If deficient, lower the pH to about 6.5 (for rockwool, about 5.7), and check that you're not adding too much P, which can lock up Fe. Use iron that's chelated for maximum availability. Read your fertilizer's ingredients - chelated iron might read something like iron EDTA . To much Fe without adding enough P can cause a P-deficiency.


Mercury contamination from dental amalgam does not lead to dramatic increases in heavy metal concentrations that would demand consequences such as weaning. Detoxification treatment is not indicated. In addition, chelating agents may mobilize heavy metals and in this way increase contamination of the mother's milk. On the other hand, amalgam fillings should only be removed in case of dental problems. Extensive restoration should be postponed until after the breastfeeding period. Wherever possible, amalgam should be avoided. However, at an individual level, the amalgam issue should not be stirred up into a toxicological crisis that puts an unjustifiable strain on the mother-child relationship.


For quantitation of tacrolimus, EDTA-anticoagulated whole blood is the specimen of choice for the same reasons provided for CsA. Whole blood samples are stable for 1 week when shipped by mail without coolant (98,99), 1-2 weeks at room temperature (99,100), 2 weeks at refrigerator temperatures (100), and almost 1 year at 70 C (100).

DNA strand cleavage

Oxidative stress by hydroxyl radical also causes direct DNA damage, mainly by strand cleavage and oxidation of pyrimidine and purine bases. DNA, because of the negative charge of its phosphate groups, acts as an anion and is therefore capable of binding many cations, including those required for Fenton chemistry, like Fe2+ and Cu+. Additionally, deoxyribose has also good iron-binding properties. This allows ''site-specific'' hydroxyl radical generation that cannot be countered by radical scavengers. Perhaps for this reason, antitumor compounds that act by DNA strand cleavage are also normally chelating agents.


Also, incubating 1 ml specimen with 10 l of alkaline phosphatase enzyme (Sigma Chemical Company, St. Louis, MO, USA) converts any fosphenytoin present in the specimen to phenytoin within 5 min at room temperature. This procedure eliminates interference of fosphenytoin in phenytoin immunoassays. The authors observed complete conversion of fosphenytoin to phenytoin by alkaline phosphatase in heparin, ethylenediaminetetraacetic acid (EDTA), and citrated plasma (16).

Parkinsons Disease

A number of polyphenols, including EGCG, function as antioxidants at low concentrations 5 . Some of their actions involve scavenging ROSs (discussed below) and induction of endogenous antioxidants via PKC 16 . ROS generation via iron dysregulation plays a role in Parkinson's disease. Toxic a-synuclein aggregates form after exposure to redox-active iron both occur within Lewy bodies, characteristic of Parkinson's disease 12 . As in Alzheimer's disease, iron also regulates degradation of IRP in Parkinson's disease. Decreased amounts of IRP lead to decreased transcription of the transferrin receptor, and thus increases in the iron transport protein, ferritin. Mice lacking the IRP gene accumulate iron in their substriatia and develop symptoms of Parkinson's disease, including tremor and bradykinesia. EGCG prevents accumulation of a-synuclein and decreases removal of IRP in murine models of Parkinson's disease 1 . Iron chelation may thus contribute to EGCG's neuropro-tective effects in...