Chelation for Coronary Heart 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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TABLE 1783 Adverse Effects of Chelating Agents

For symptomatic patients without encephalopathy, and for asymptomatic patients with elevated PbB levels requiring chelation, the use of BAL or DMSA (see discussion below) with or without CaNa2-EDTA and the dosing schedules are determined by the PbB levels, the presence or absence of symptoms, and changing practice as more experience with DMSA is obtained. Treatment may be initiated in the ED. Children who are symptomatic but not encephalopathic can be treated as discussed earlier, except with doses of BAL, 50 mg m2, and CaNa2-EDTA, 1000 mg m2 per 24 h, or with DMSA plus EDTA. A retrospective study of 45 children with lead levels of 45 pg dL or more treated either with BAL plus EDTA or DMSA plus EDTA found comparable reductions in lead levels, with fewer side effects in the DMSA group.10 The Centers for Disease Control and Prevention (CDC) recommend immediate chelation therapy for children with blood lead levels of 70 pg dL or more.8 Symptomatic, nonencephalopathic adults may be...


Chelation is the process whereby an organic moiety acts as a ligand to bind a metal ion through two or more coordination bonds. Some low-molecular-weight compounds that may be released during the digestion of food can act as metal chelators and increase metal solubility in the intestinal lumen. In some circumstances, chelated forms of metals are naturally present in food, such as heme iron (part of hemoglobin or myoglobin protein) found in meat. Heme is a stable protoporphyrin ring-containing compound that protects a central iron atom from interacting with other potentially deleterious compounds, such as phytic acid, that would reduce its availability and inhibit iron absorption. Nonheme iron bioavail-ability is affected by various enhancing and inhibitory substances found in food. In contrast, heme iron bio-availability is not. The heme moiety is absorbed intact by the enterocyte. Inside the enterocyte a cytosolic enzyme heme oxygenase breaks apart the protopor-phyrin ring and...

The Immobilisation Degradation or Monitoring of Pollutants from a Biological Origin

Immobilisation can be achieved by chemicals excreted by an organism or by chemicals in the neighbouring environment which trap or chelate a molecule thus making it insoluble. Since virtually all biological processes require the substrate to be dissolved in water, chelation renders the substance unavailable. In some instances

Detection Of Saliva Stains

EDTA and Extracted DNA is typically stored at 20 C, or even 80 C for long-term storage, to prevent nuclease activity. Nucleases are enzymes (proteins) found in cells that degrade DNA to allow for recycling of the nucleotide components. Nucleases need magnesium to work properly so one of the measures to prevent them from digesting DNA in blood is the use of purple-topped tubes containing a blood preservative known as EDTA. The EDTA chelates, or binds up, all of the free magnesium and thus prevents the nucleases from destroying the DNA in the collected blood sample.

Depletion of abundant proteins followed by fractionation of intact proteins

Misek et al. 25 identified many isoforms and compared relative abundance of proteins in serum, EDTA-plasma, and citrate-plasma labeled, respectively, with the fluorescent dyes Cy3, Cy5, and Cy2 after top-6 immunoaffinity depletion. The three labeled, depleted samples were subjected to three-dimensional protein fractionation by pi, hydrophobicity, and Mr About 3000bands on 1-D SDS gels with two-fold differences in intensity of fluorescence in dye pairs were excised and analyzed by MS MS, yielding a total of only 82 non-redundant proteins 28 proteins were identified in ten or more different fractions. Complement C3 and clusterin are presented as examples of proteins whose biologically significant cleavage products can be identified with this method. Not surprisingly, the yield in MS MS was greater for proteins with higher intensity (abundance). Multiple isoforms reduce the concentration of a protein in any particular spot or fraction and may react very differently with antibodies used...

Effects of lysozyme in real food product trials

Akashi and Oono (1972) reported lysozyme to exert a weakly preservative effect in lightly salted fish, when lysozyme was employed as a dipping treatment in 1 gelatin-0.05 lysozyme solution, but treatment with sorbic acid consistently resulted in a better preservative effect than lysozyme in these fish products (Akashi and Oono, 1972). Later reports have confirmed that lysozyme exerts only weak antibacterial potency in animal products such as pork sausage (bratwurst) and Camembert cheese (Hughey et al., 1989). In Camembert cheese lysozyme by itself or together with EDTA reduced an inoculated L. monocytogenes population by ten-fold during the first 3-4 weeks of the ripening period, but the effect of lysozyme decreased with longer storage, where L. monocytogenes was found to grow unhindered in the artificially inoculated, lysozyme-containing cheeses (Hughey et al., 1989). A lysozyme dip treatment (3 mg mL) of cod fillets spiked with L. monocytogenes resulted in retarded growth of...

Analysis with antibody arrays

Fig. 4 Comparison of twenty blood samples collected into tubes having either PI cocktail and EDTA (blue circles) or EDTA alone (red squares). Samples represent time zero timepoints, processed from whole blood into plasma, aliquoted, and frozen, within 15 min from blood draw. Upper graph 20data points for each tube type, showing m z ratio of approximately 6803 and signal intensity, after Fig. 4 Comparison of twenty blood samples collected into tubes having either PI cocktail and EDTA (blue circles) or EDTA alone (red squares). Samples represent time zero timepoints, processed from whole blood into plasma, aliquoted, and frozen, within 15 min from blood draw. Upper graph 20data points for each tube type, showing m z ratio of approximately 6803 and signal intensity, after

[2 Phenotype Changes of Fut8 Knockout Mouse Core Fucosylation Is Crucial for the Function of Growth Factor Receptors

For preparation of embryonic fibroblasts, a whole mouse embryo at 18.5 days after coitus was dissected, and the head and all internal organs were removed. The carcasses were minced, incubated in PBS (-) containing 0.05 trypsin, 0.53 mM EDTA, and 40 g ml DNase at 37 for 30 min with stirring three times, and then cells were plated on a 100-mm dish in

Advantages and disadvantages of the NMR technique

The measurements are easily repeatable and reproducible over the long term. The instrumental parameters that need careful attention from the analyst and that may vary from one data recording session to another are the tuning and the resolution. A difference (even small) in tuning will affect the intensity of all peaks in the spectra. In order to compare quantitatively sets of spectra recorded at different times, all the spectra can be normalised to the reference peak provided that a fixed amount of internal standard is added to all the samples. The problems of repeatability and reproducibility of chromatographic techniques that are caused by the ageing of columns (affecting separation performances and changing retention times), temperature fluctuations, etc., are not applicable to NMR. The disturbances that can occur are the small shifts of the NMR peaks caused by differences in pH in the NMR spectra of food extracts or the broadening of certain signals due to chemical exchange of...

Electrophoretic Karyotyping

For the determination of the number and size of the chromosomes by electrophoresis, it is necessary to keep the chromosomal DNA molecules intact during the procedure of cell and chromosome dissolution and the subsequent electrophoretic separation. Therefore, the cells or spheroplasts are embedded in agarose, which protects the DNA against mechanical breakage while allowing the free diffusion of solutions necessary for lysis and digestion. To inhibit DNAses, all treatments are carried out at high concentrations of EDTA. To release the DNA from the chromosomes, the embedded cells or spheroplasts are treated with Proteinase K, which degrades most of the chromosomal proteins. 2. Determine cell density by either cell chamber (hemocytometer) counts or photometrically. Flocculation or poor separation of cells may cause a problem in cell counting. This can usually be overcome by one or two washes with 50 mM EDTA. 1. Centrifuge a sample of the culture that contains approx 5 x 109 cells. Wash...

Deferoxamine Challenge Test

This test is sometimes performed in cases where toxicity is not clear and laboratory results are still pending (1st 6 h). The patient is given deferoxamine 50 mg kg up to 1 g IM, and all the urine is collected. If the urine turns a vin rose color, the test is positive and chelation is continued intravenously. However it is important to note that the urine color change is an insensitive marker and that a negative challenge test does not rule out the presence of iron toxicity. For that reason, many toxicologists no longer recommend this test.

Dna Separation Methods Slab Gel And Capillary Electrophoresis

Capillary Electrophoresis Dna

Electrophoresis buffer is poured over the gel until it is fully submerged. Two buffers are commonly used with electrophoresis, Tris-acetate-EDTA (TAE) and Tris-borate-EDTA (TBE). Samples are mixed with a loading dye and carefully pipetted into each well of the submerged gel. This loading dye contains a mixture of bromophenol blue, a dark blue dye which helps to visually see the sample, and sucrose to increase the sample's viscosity and help it stay in the well prior to turning on the voltage and initiating electrophoresis.

Quantitative Nasba And Tma Applications

HIV-1 RNA Using the NucliSens HIV-1 QT (bio-Merieux), 13'14 between 25 and 5 x 106 copies mL of HIV-1 RNA can be detected with a 95 detection rate of 176 copies mL, by using 1 mL of EDTA, citrate, or heparin plasma as a sample. Murphy et al. 13 compared the Roche COBAS AMPLICOR MONITOR version 1.5 (Roche Molecular Systems), the NucliSens HIV-1 QT with extractor, and the Bayer Quantiplex Version 3.0 (Bayer) for quantification of HIV-1 RNA in 460 plasma specimens from HIV-1 infected patients. They observed that NucliSens showed 100 specificity and the best sensitivity with an input of 2 mL. In another recent study, the same specificity was displayed by the NucliSens HIV-1 QT assay. 14 In a comparison with the ultrasensitive AMPLICOR HIV-1 Monitor version 1.0 assay (Roche Molecular Systems), the NucliSens and the AMPLICOR assays were equivalent in detecting HIV-1 RNA (concentrations of 103-105 copies mL), although at lower RNA concentrations, the NucliSens assay was more sensitive.

Preanalytical Variables Anticoagulant

Accurate assessment of in vivo or in vitro cellular expression of molecules requires optimal preanalytical conditions to prevent in vitro artifactual activation. The choice of anticoagulant is one of the critical preanalytical conditions because anticoagulants exert different effects on the activation of cells ex vivo. Historically, sodium citrate has been the favored anticoagulant for use in the studies of platelet activation and function, including aggregation and adherence. However, recent studies have shown that the anticoagulant CTAD (a mixture of sodium citrate, theophylline, adenosine, and dipyridamole) is better for retaining the ex vivo status of platelets (80). But it should be noted that this anticoagulant is light-sensitive and, when exposed, stable for only up to 4 h (81). In addition, when CTAD is combined with EDTA (ethylenediaminetetraacetic acid) and the blood held at 4 C, platelet activation after venesection is inhibited for at least 6 h after venesection (82,83)....

Analysis of Platelet Surface Activation Antigens aIIbfi3

The first antibody that recognized the activated conformation of the molecule glycoprotein OjIb P3 was developed in 1994. This antibody, PAC-1, is an IgM antibody that recognizes a sequence on the oIIb P3 molecule that is exposed on activation. It may not be used with samples that have been fixed with formaldehyde (85). The normal conformation of OjIb P3 is dependent on the presence of divalent ions (predominantly Ca2+) at physiological concentrations (86). Removal of divalent ions results in an altered conformation, and therefore blood samples in EDTA are unsuitable for study of the activated form of this molecule. EDTA may in itself cause dissociation of the OjIb P3 complex (69).

Organic acids mode of action

The principal mode of action of organic acids is the release of protons from carboxylic groups, thus lowering the pH of their environment. Proton release means dissociation of the acid molecule, with the degree of dissociation depending on the pKa value. Strong acids tend to be fully dissociated and thus produce a strong drop in pH, while weak acids are only partially dissociated. This has two important consequences (a) organic acids with two or three carboxylic groups react on changes of pH by changes in protonation of their carboxylic groups, i.e., they may 'buffer', and (b), in their undissociated state, some of these acids can pass through lipophilic cell membranes. Consequently, on a molar basis, the antibacterial efficacy of weak acids is stronger than that of strong acids. Table 5.3 reviews properties of selected organic acids. Kation effects are reported also. Protons may change the conformation of polar membrane proteins. Binding metal ions (Fe3+, Cu++, Mn++, Ca++, Mg++) by...

Cell Cultures of Coelomocytes New Tools to Detect Marine Pollution

As already mentioned, the development of in vitro cultures of coelomocytes has been considered important and recommended in order to better analyze the mechanisms involved in defence stress response and to overcome the stress of manipulation. Media for keeping sea urchin coelomocytes outside of the animal, for a reasonably sufficient period of time, generally involve the use of anti-coagulant reagents, such as the chelating agents EDTA and EGTA. In our studies, P.lividus coelomocytes were kept and separated into subpopulations by gradients in an EDTA-containing buffer (ISO-EDTA). This medium was good enough to show, for the first time, that coelomocytes respond to physical and chemical stresses by elevating the hsp70 expression levels (Matranga et al. 2000). However, it was not preserving the morphological features of freshly collected cells and it was not usable at all for in vitro exposure

Reagents and Solutions

Point Care Flow Cytometer

LB01 15 mM Tris 2 mM Na2EDTA 0.5 mM spermine.4HCl De Laat's 15 mM HEPES 1 mM EDTA Na2.2H2O 0.2 (v v) a Final concentrations are given. MOPS, 4-morpholinepropane sulfonate DTT, dithiothreitol Tris, tris-(hydroxymethyl)-amino-methane EDTA, ethylenediaminetetraacetic acid HEPES, acid. For details on the buffer preparation see the original reference(s). b pH of the buffers is not adjusted. Isolation buffers, in addition to releasing nuclei from the cytoplasm in sufficient quantities, must also maintain nuclear integrity throughout the experiment, protect DNA from degradation by endonucleases and permit stoichiometric DNA staining. From about 26 different isolation formulas described, six are commonly used in plant DNA flow cytometry (Loureiro et al. 2006a Table 4.3). Their usual components include (i) organic buffer substances (e.g. Tris, MOPS and HEPES) to stabilize the pH of the solution (usually set between 7.0 and 8.0, which is compatible with common DNA fluorochromes) (ii) non-ionic...

The Importance Of Determining Pon1 Status

Variability of PON1 protein levels within each genotype (Fig. 2). Indeed, PON1 levels are at least as important as genotype in determining rates of clearance of oxidized lipids, as well as determining resistance to organophos-phate toxicity. 10 By plotting the rates of diazoxon hydrolysis against paraoxon hydrolysis at high salt concentration (2 M NaCl) for individuals in a population, the subjects are clearly divided into three groups individuals homozygous for PON1Q192, heterozygotes (PON1Q R192), and individuals homozygous for PON1r192 in addition, information of PON1 activity levels is also obtained (Fig. 2 Ref. 9 ). The procedure can be easily carried out in 96-well microtiter plate readers capable of following continuous enzyme kinetics at the appropriate wavelength (for details, see Ref. 14 ). For this assay, plasma from blood collected in heparin or citrate can be used, but not ethylenediaminetetraacetic acid (EDTA) plasma, because EDTA irreversibly inhibits PON1. The accuracy...

Quantification and Characterization of Platelet Microparticles

Platelet Microparticles

Blood (10 pL) preferably anticoagulated with EDTA CTAD is incubated for 5 min at 4 C with FITC-isotype control (2l) and PE-isotype control (2l) or with FITC-CD42a (2l) and PE-CD62P (2l). Samples are diluted to 1 mL with HBSS-BSA containing LDS-751 (as above). Unlabeled polystyrene spheres 1.09 m in diameter (at 1 X 10 6 from the stock solution supplied Sigma-Aldrich) and EDTA CTAD at 0.25 of the concentration used for preventing blood coagulation are added and analyzed immediately by flow cytometry.

Therapeutic modification of the immune response

For example, the use of F(ab)2 fragments of 'humanized' mAbs may abrogate the nonspecific binding and host response problems. Improved chelation techniques may decrease the extent of complex dissociation. Other areas of research include the use of mAbs linked to enzymes which are then used to activate an inactive prodrug - antibody-directed enzyme prodrug therapy (ADEPT). This approach has the additional advantage that the half-life of specific binding is usually much greater than that of nonspecific binding, thereby improving the therapeutic ratio as administration of the active drug can be delayed to allow clearing of such nonspecifically bound antibody. By means of such approaches, it may become possible in the future to utilize mAbs for systemic therapy, probably as an adjunct to more conventional debulking treatment such as surgery, radiotherapy or chemotherapy although the clinical utility of ADEPT remains to be convincingly demonstrated.

Evaluation and Supportive Care

Iron poisoning is a clinical diagnosis. Patients who arrive at an emergency department asymptomatic and with normal findings on physical examination, and who remain so for 6 h following ingestion, do not require specific medical treatment for iron poisoning. Patients who are symptomatic from their ingestion should first be stabilized with attention to airway, breathing, and circulation, after which GI decontamination and chelation therapy may proceed. Dialysis is not effective in clearing iron.

Measurement of Reactive Oxygen Species ROS Production

The detailed protocol of this method is as follows. Experimental animals are wrapped in a towel to absorb external seawater and then bled by cutting the tip of one arm and draining 3 ml of coelomic fluid in an equal volume of sterile anticoagulant buffer Ca2+, Mg2+-free artificial seawater (CMFASW) (Noble 1970) with 1.2x10-2M EDTA at 4 C. The amoebocyte concentration of this suspension is determined as follows 200 l of the cell suspension from each animal is distributed in wells of a UV-transparent microplate (96-wells, UVstar, Greiner), cells are mildly agitated for 5 s and the absorbance at 280 nm is determined using a microplate reader (Spectrafluor Plus, TECAN three flashes per well). This is related to the cell concentration by using the following predetermined relation (Coteur et al. 2002) A280 0.157 x cell concentration (x106 cells ml-1)+0.067. The suspension is then centrifuged (400 g, 10 min, 4 C) and the supernatant replaced by CMFASW (without EDTA), the volume of which is...

Biological Removal Biotransformation and Biosorption of Metal Ions from Contaminated Wastewater

Metal ion contaminants in wastewater can be removed by microorganisms by either a direct or indirect influence on the redox state of the metal ions or through biosorption of metal ions on the cell surface (Lovley and Coates, 1997). Some microorganisms have also developed resistance mechanisms against toxic metals by changing the oxidation state without supporting anaerobic growth. Certain bacteria, yeasts, fungi, and algae can actively accumulate intracellular metal ions against a gradient. The process of bioaccumulation of metal ions depends on living, metabo-lically active cells, whereas biosorption is a passive, energy-independent process that can be mediated also by inactive cell material. Biosorption of metal ions includes mechanisms such as ion exchange, chelation, matrix entrapment, and surface sorption (Unz and Shuttleworth, 1996). After biosorption or active removal of metal ions Changing the redox state of natural and anthropogenic radionuclides by metal-reducing...

Dietary Management of Secondary Overnutrition

Chelating agents in Wilson's disease and recurrent phlebotomy in hemochromatosis. In a related variant condition, African hemosiderosis, common among Bantu in southern Africa, removing concentrated iron sources from the diet, specifically the iron-loaded native beers, provides effective long-term control.

Role Of Rocket Immunoelectrophoresis In Food Allergens Analysis

Figure 1 Crossed immunoelectrophoresis of serum from a patient with chronic urticaria and angioedema. The separating electrophoretic step was done in the presence of Ca2', and the second electrophoretic step was performed with a mixture of anti-C1s and anti-C1r in the gel and with (A) EDTA or (B) Ca2 in the buffer. (Reproduced with permission of Georg Thieme Publishers, Stuttgart, from Opferkuch W, Rother K and Schultz DR (eds) (1978) Clinical Aspects of the Complement System.)

Biochemical Mechanisms For Ironfolate Interactions

Clinical manifestations of folate deficiency during iron deficiency can occur in the absence of depleted tissue folate (17). This observation indicates that catabolism-mediated folate deficiency cannot, in itself, fully account for the iron-folate relationship. Iron status is influenced by three pools of cellular iron the functional pool that is bound by iron-requiring proteins, the storage pool that is ferritin-bound, and the labile, or regulatory, pool that exists free in solution. Elevated expression of HCF in the absence of increased iron availability lowers intracellular regulatory iron concentrations and triggers the cellular iron-deficiency response, which includes increased expression of the transferrin receptor and increased rates of iron uptake (51). A recent study demonstrated that increased expression of HCF, but not increased LCF, in cell cultures alters the relative distribution of folate one-carbon-substituted cofactors, indicating that HCF expression alters the flux of...

Combining traditional and new preservation techniques

The use of high pressure in combination with mild heat is promising (Patterson, et al., 1995a). This strategy is successful because there is evidence that microbial injury can occur at significantly lower pressures than that required for inactivation (Patterson et al., 1995b). E. coli cells surviving pressurization become sublethally injured and develop sensitivity to physical and chemical environments to which the normal cells are resistant (Kalchayanand et al., 1998a Hauben et al., 1996). This suggests that exposing E. coli to a combination of different intervention strategies renders the bacterium sublethally injured and serves as an effective food preservation method. Hauben et al. (1996) assessed the destruction and sublethal injury of E. coli by hydrostatic pressure and by combinations of high pressure treatments with lysozyme, nisin, and or EDTA. High pressure treatments (180 to 320 MPa) disrupt the bacterial cells outer membrane, causing periplasmic leakage and sensitization...

Outline Of The Method

The comet assay can be conducted on virtually any nucleated cell that can be brought into suspension. Thus, both cultured or freshly isolated cells from any organ can be used. Isolating cells from solid tissues usually involves time-consuming procedures, often implying the use of extracellular matrix-digesting enzymes, which can damage DNA. Instead, for the study of endogenous levels of DNA damage, it is necessary to use rapid isolation techniques to reduce the onset of artefacts, and, for some purposes, intact nuclei isolation by simple homogeniza-tion is advisable. 5 International expert guidelines have been provided for using the assay in in vivo genotoxicity testing. 6 Care should also be taken to prevent DNA damage induction during the procedure. For this purpose, it is convenient to work in dim light to avoid UV-induced DNA damage, and all solutions used should contain EDTA to inhibit endonuclease activity. NaCl, 100 mM Na2EDTA, 1 Triton X-100, and 10 dimethylsulfoxide....

Possible Mechanisms Of Anticancer Effects

Plaatje Continue Verbetering

Also due to its structure, PA can chelate polyvalent cations, such as iron, calcium and zinc 83 Figure 14.3(C) . These minerals are required for vital cellular functions, and their chelation may affect cellular growth kinetics. The chelation of minerals vital to DNA synthesis and cell growth may be a factor in studies that have shown a decrease in cell proliferation with PA supplementation 13,17 . This may also be a mechanism in the protective effects of PA on human rhabdomyosarcoma 59 and HepG2 liver cancer cells in athymic mice, where PA was injected directly into the tumor. In fact, studies have shown that PA can reduce increased colonic cell proliferation 15 and inhibit colon tumorigenesis 33 induced by the addition of minerals such as iron to the diet. Because the iron necessary to catalyze hydroxyl radical production may be diet derived 88 , its chelation by PA may inhibit hydroxyl radical production and subsequent oxidative damage Figure 14.3(C) . In fact, individuals consuming...

Activators and Inhibitors

Many compounds inhibit PPO activity but only a limited number of PPO inhibitors are considered acceptable on grounds of safety and expense for use to control enzymatic browning during food processing. Practical aspects of browning inhibition are discussed later. There are two principal types of PPO inhibitor compounds that interact with the copper in the enzyme and compounds that affect the site for the phenolic substrate. The metal-chelating agents such as cyanide, diethyldithiocarbamate, carbon monoxide, dimercaptopropanol, sodium azid, phen-ylthiourea, and potassium methyl xanthate are all inhib itory. In general, there are five major types of PPO inhibitors based on their chemical characters (56,57) (1) reducing agents, such as ascorbic acid and sulfur dioxide (2) copper-chelating agents, such as carbon monoxide, tropolone, and diethyl dithiocarbamate (3) quinone couplers, such as cysteine and glutathione (4) substrate analogues, such as cinnamic acid, p-coumaric acid, and benzoic...


The composition of the Cl complex of the complement system can be demonstrated by crossed Immunoelectrophoresis with monospecific antibodies against the subcomponents Clq, Clr and Cls, respectively. When fresh normal serum was run with Ca2 present in both electrophoretic steps, the native Clqrs complex remains at the application site and no visible precipitate appeared with anti-Clq, anti-Clr or anti-Cls in the second gel. When the second electrophoretic step was done with EDTA present, two separate precipitation arcs appeared at the appli Crossed immunoelectrophoresis has been used to study Cl and Cl activation in various diseases with an immunological background. Figure 1 shows the findings with serum from a patient with chronic urticaria and angioedema. Electrophoresis was run with monospecific antisera against the Cl subcomponents and with Ca21 present in the first step and with EDTA in the second step. Two separate precipitation arcs appeared at the application site, representing...

Heavy Metal Intoxication

Ingestions of one of several heavy metals can lead to systemic manifestations. Lead poisoning, or plumbism, is the most common heavy metal poisoning. Systemic signs of lead poisoning are highly variable depending on the age of the patient and the amount of lead ingested. Symptoms range from colic, iritability, fatigue, and anemia to encephalopathy. Intraorally, lead poisoning presents as an ulcerative stomatitis or a bluish hue to the buccal mucosa. The classic bluish lead line on the ginigiva, secondary to subepithelial deposits of lead sulfide, also may be seen. In addition, a tremor on tongue thrusting, excessive saliva production, metallic taste, and severe periodontal disease may occur. Treatment is chelation therapy.40

Linkage of MS data and antibodybased measurements

The preparation method that generally gave the highest protein concentrations varied among the four data sets. The GNF and MSI sets had higher protein abundances in the EDTA-plasma preparation, the DB set had higher abundances in the serum preparation, and the VARI set had highest values in the serum and heparin-plasma. The GNF and MSI sets focused on cytokine detection, and the relatively higher concentration of the cytokines in EDTA-plasma could indicate a protective effect of EDTA on cytokine stability, perhaps through EDTA's role as a protease inhibitor. The more abundant, common serum proteins measured in the other two sets could be less susceptible to protease activity and therefore not necessarily higher in the EDTA-plasma preparation. Other sources of variation in concentration could be the anticoagulant-induced release of certain analytes by lymphocytes, such as the release of tumor M2-PK in heparin-plasma but not in EDTA-plasma The analysis of specific proteins showed that...

TABLE 514 Common Causes of Restrictive Cardiomyopathy

TREATMENT AND DISPOSITION Symptoms and signs of CHF, particularly right-sided failure, with a normal-size cardiac silhouette on chest x-ray should prompt a suspicion of underlying restrictive cardiomyopathy, constrictive pericarditis, or diastolic LV dysfunction (most commonly due to ischemic heart disease, hypertension, or age-related changes in ventricular diastolic compliance). Doppler echocardiographic studies and cardiac catheterization with hemodynamic assessment are often required to differentiate between the above-mentioned entities. Computed tomography and magnetic resonance imaging of the heart have also been shown to be of value in differentiating constrictive pericarditis and restrictive cardiomyopathy. 11 Timely diagnosis is important because constrictive pericarditis can be surgically corrected and diastolic LV dysfunction not due to restrictive cardiomyopathy usually responds well to drug therapy (b blockers or calcium-channel blockers). The medical management of...

Biocompatibility and Toxicity

Besides local toxicity, discussed above, there are numerous other modes of potential adverse interactions involving excipients (19,20). Many of these pose little threat provided the amounts of excipients are constrained to certain levels. Excessive amounts, however, can cause problems, particularly for patients who are intolerant of even modest levels. Commonly used phosphate buffers may cause calcium loss with formation of insoluble calcium phosphates when such buffers are administered in over-ambitious amounts (21). Calcium phosphate precipitation has been noted particularly in nutritional parenteral admixtures for neonates because of the high nutrient requirements. Similarly, renal toxicity has been associated with depletion of zinc and other trace metals caused by large parenteral doses of ethylenediaminete-traacetic acid (EDTA) (22). Excessive absorption of glycine solutions, when used as irrigants during transurethral resections, can cause hyponatremia, hypertension, and...

Drug interactions affecting absorption

Chelation and Adsorption Drugs may form insoluble complexes by chelation in the gastrointestinal tract. Chelation involves the formation of a ring structure between a metal ion (e.g., aluminum, magnesium, iron, and to a lesser degree calcium) and an organic molecule (e.g., anti-infective medication), which results in an insoluble compound that is unable to permeate the intestinal mucosa because of the lack of drug dissolution.

Oxidative Stress Antioxidants And Lipid Peroxidation

These investigators also found that enhanced lipid peroxidation was associated with worsened neuropathy in diabetic rats. In another study, this group found that the antioxidant, a-lipoic acid, which improves nerve function and blood flow when administered to diabetic animals (Cameron et al. 1998), abolished lipid peroxidation induced in vitro by ascorbate-iron-EDTA or when the nerve was incubated in 20 mM glucose (Nickander et al. 1996). Van Dam et al. (1998) reported that malondialdehyde levels in diabetic rats were elevated in plasma, but not in the nerve. These investigators found, in contrast, that nerves of vitamin E-deficient rats had a markedly increased content of malondialdehyde. It has been shown that carboxymethyllysine, the major epitope recognized by antibodies against chemically modified proteins, may be formed not only via glycoxidation of glucose but also during lipid peroxidation of polyunsatu-rated fatty acids (Fu et al. 1996). Taken together, these...

Management of Thalassemia Major

Patients affected by thalassemia major are treated with regular blood transfusions and iron chelation therapy with desferrioxamine B (DFO). Life expectancy with this treatment extends to the third decade. The alternative oral iron chelator deferiprone (L1) is indicated only in patients with proven allergy or toxicity from DFO. Alternative chelation strategies and drugs, including the combination of deferiprone and DFO or ICL670 alone, are under investigation. Bone marrow transplantation (BMT) or cord blood transplantation from HLA-identical siblings represents an alternative to traditional transfusion and chelation therapy. In patients without liver fibrosis and low level of iron accumulation, the disease-free survival is over 90 . However, BMT from HLA-identical but unrelated donors has a lower (60 ) disease-free survival. 4-6

Practical Toxicity Issues

In broad terms, type-A metals are less toxic than type-B, but this is only a generalisation and a number of other factors exert an influence in real-life situations. Passive uptake by plants is a two-stage process, beginning with an initial binding onto the cell wall followed by diffusion into the cell itself, along a concentration gradient. As a result, those cations which readily associate with particulates are accumulated more easily than those which do not. In addition, the presence of chelating ligands may affect the bio-availability and thus, the resultant toxicity of metals. Whereas some metal-organic complexes (Cu-EDTA for example) can detoxify certain metals, lipophilic organometallic complexes can increase uptake and thereby the functional toxic effect observed.

Magnetic resonance scan

Chromium ethylenediaminetetraacetic acid clearance Ethylenediaminetetraacetic acid (EDTA) labelled with a radioactive an isotope of chromium is given intravenously and measurement of blood and urine concentration gives a close approximation to the GFR. The agent is filtered at the glomerulus only, with little or no tubular secretion occurring. It therefore provides a quick and convenient test of GFR.

TABLE 1998 Commonly Treated Forms of Internal Contamination

Chelating agents such as calcium and zinc salts of diethylenetriamine pentaacetic acid (Ca-DTPA and Zn-DTPA, respectively) are effective treatments for contamination with heavy metals and rare earths that emit alpha radiation. If alpha-emitting contamination is detected in wounds or in the nares or oropharynx, treatment with DTPA should be initiated promptly, ideally within 1 to 2 h after contamination has occurred. Potential contraindications for the use of DTPA are severe renal dysfunction, thrombocytopenia, or leukopenia. DTPA is administered systemically by slow IV push or by aerosol administration. Ca-DTPA has been shown to be more effective in animal studies and is the preferred form of drug for the initial one to two days of treatment. Zn-DTPA is less toxic and recommended for treatments of longer duration and of pregnant females. For aerosol administration, Ca-DTPA is preferred because of the metallic taste associated with Zn-DTPA. Most DTPA solutions in nuclear medicine...

Internally Contaminated Patients

DECORPORATION TREATMENT Once the radioactive material crosses into the extracellular fluid, incorporation has occurred and elimination is more difficult. Methods of decorporation include blocking agents, isotopic dilution, displacement, mobilizing agents, and chelation. Treatment with blocking agents reduces the uptake of a radioisotope at an organ or metabolic site by saturating the site with a stable form of the isotope. Isotopic dilution therapy involves administering large quantities of a stable form of the isotope, thus diluting the concentration of the radioisotope. In displacement treatment, a different but similar stable element is administered that will act as a competitor with the radioisotope for an uptake site. Mobilizing agents induce body tissues to release radioisotopes by increasing the natural turnover process. Chelating agents are organic compounds that provide an ion exchange matrix. The exchanging of inorganic ions results in stable nonionized ring complexes that...

Storage Of Dna Extracts

Traditionally, purified DNA is stored in either sterile water or TE (10 mM Tris, pH 7.5-8.0, 1 mM EDTA). In either of these solutions, DNA may be stored for several months at either 4 C or 20 C with little degradation. Extracted DNA stored in TE at temperatures between 70 C and 37 C for 6 months demonstrated little degradation, whereas DNA stored at 45 C to 65 C shows signs of degradation within 24 hr and was completely degraded by 8 days. 37 Repeated freezing and thawing of DNA does not appear to result in degradation. For longer cryogenic storage, DNA should be stored in nuclease-free solutions at 70 C (ultrafreezers) to 196 C (liquid nitrogen). Alternatively, DNA may be precipitated and stored in ethanol often layered with chloroform at room temperature or 4 C. Although lyophilized DNA stored at 20 C is protected from degradation, problems with rehydration after prolonged storage have been reported. 37 Buffers containing EDTA have been used to store DNA at room temperature with...

Selected nutrient deficiencies

A deficiency of iron is the most common nutritional problem worldwide, even in industrialized countries. On the one hand, free iron is necessary for bacterial growth (removal of iron with the help of lactoferrin or other chelating agents reduces bacterial multiplication, particularly in the presence of specific antibody) and, on the other, iron is needed by neutrophils and lymphocytes for optimal function. The lymphocyte proliferation response to mitogens and antigens is impaired response to tetanus toxoid and herpes simplex antigens is low in iron-deficient subjects and iron therapy results in a significant improvement in their response. There are many molecular explanations for impaired lymphocyte and neutrophil function in iron deficiency, including the deficiency of myeloperoxidase and ribonucleotidyl reductase.

Preliminary separation steps

It is advisable to maintain the ionic strength of the medium to aid protein stability. Many other additives can be included at this stage, e.g. protease inhibitors, or EDTA to chelate metal ions which can catalyse oxidation. However, with prior knowledge of the protein stability these can be kept to a minimum as they will inevitably have to be removed at a later stage and may cause unnecessary complications and delay. As a general rule, the preliminary extraction should be performed quickly, at sub-ambient temperatures.

Factors that improve the efficiency of lysozyme

The combination of lysozyme with other substances notably with EDTA, glycine, salt (NaCl), or other preservatives such as sorbate or ethanol have shown promise (Proctor and Cunningham, 1988). Use of lysozyme together with other antimicrobial enzymes or in combination with physico-chemical stressing of the cells, such as decreased pH and temperature, also enhance the bacteriocidal effects of lysozyme (Johansen et al., 1994 Proctor and Cunningham, 1988). Several approaches have been attempted in order to render hen egg white lysozyme more active against Gram-negative bacteria. EDTA addition, in particular, has been claimed - and widely investigated - to increase the permeability of the outer membrane, and thereby increase the accessibility of lysozyme to the peptido-glycan of Gram-negative organisms (Proctor and Cunningham, 1988). The exact mechanism by which EDTA may destabilise the outer lipopolysaccharide layer and perturb the membrane has not been fully clarified, however. With...

Synthetic Oxygen Carriers

Normally only 4 of the oxygen delivered is carried in solution (0.0225 ml 02 100 ml blood kPa), the rest is carried as oxyhaemoglobin. To rely entirely on dissolved oxygen (if this could all be extracted) would require either an increased cardiac output to 13 l min or an inspired oxygen pressure of 2-2.5 bar. The latter is the principle behind the treatment of carbon monoxide poisoning but requires hyperbaric equipment and risks oxygen toxicity. Therefore, an oxygen carrier is required that will carry oxygen more efficiently than plasma or plasma substitutes, and that will release it to the tissues. There are two options perfluorocarbons and chelating agents.

Collection of Deoxyribonucleic Acid from Blood Samples

It is easiest to amplify DNA that will be used for genetic testing when the DNA is taken from blood samples.910 Ideally, at the time of autopsy the coroner or pathologist collects 15 ml of blood in several tubes containing ethylenediaminetetraacetic acid (EDTA), which prevents coagulation and degradation of the DNA. The tubes are stored at 4 C until the DNA is extracted for analysis, which should be within 1 week, although sometimes we have extracted DNA 4 months after collection. If the blood samples are collected in tubes that do not contain an anticoagulant, the DNA should be extracted promptly (within days of the initial collection).

Biotechnological Potential of MCT

Matrix Model Trotter

Components may be isolated from the connective tissue and then reassembled with other biological or synthetic elements to form a novel composite. Examples of biomaterials produced in this way are Integra a combination of bovine collagen, shark chondroitin-6-sulphate and a silicone sheet (the last acting as an artificial epidermis), which is used as a skin substitute (Ramos-e-Silva and Ribeiro de Castro 2002), and acellular blood vessel grafts that consist of intestinal submucosa and bovine collagen (Huynh et al. 1999). It is as a source of such components that MCT could make the most direct contribution to biomaterial design, largely by virtue of the extractability of its collagen fibrils. It is notoriously difficult to extract intact collagen fibrils from the post-fetal connective tissue of vertebrates (Trotter et al. 1997). For this reason, the collagen used in existing reassembled biomaterials is in the form of disaggregated molecules. These have the advantageous property of...

Homozygous B Thalassaemia Thalassaemia Major Cooleys Anaemia

There is absent or greatly reduced P chain synthesis. This becomes apparent with the natural fall in foetal haemoglobin (a2 y2) levels due to the switch from y to P chain production. The infant presents with anaemia during the first 6 months of life. If transfusion therapy is not instituted then the infant may demonstrate the typical thalassaemic facies (frontal bossing, maxillary hyperplasia). With transfusion therapy there is normal development for the first decade, but iron overload then becomes clinically apparent in the absence of chelation therapy (desferoxamine) and death occurs in the second or third decade. Chelation is usually started in infancy and with good compliance patients can expect to live well into their 30's and perhaps longer. In a select group of children bone marrow transplantation is an option, but not without risk of transplant-related death.

Availability of Fe in natural waters

Some organisms, such as blue-green algae, have increased their ability to scavenge Fe by the production of extracellular ferric-specific chelating agents or 'siderophores' (Wilhelm et al., 1996). These low MW compounds function as extracellular ligands which aid in the solubilization and uptake of Fe3+ in environments where the low availability of Fe3+ serves to limit growth. The production of sidero-phores by terrestrial microorganisms and their role in iron-acquisition have been well documented for bacteria and fungi (Winkelmann, 1991), but are less clear for blue-green algae and their part in aquatic iron cycles.

Consistent alterations in specific protein abundances

We then examined whether specific proteins, as opposed to all the proteins in general, were consistently highest or lowest in a certain preparation type. Evidence for such a bias would be indicated by multiple specimen sets showing agreement in the alteration of the concentration of a specific protein, e.g., if all three of the BD specimen sets showed a certain protein higher in a certain preparation method. We identified the proteins that always gave a highest value in one particular preparation method, in every specimen set, and in every replicate experiment. Such biases toward a particular preparation method are more than 99 likely not to have occurred by chance, as determined by a permutation test similar to that described above. A summary of these results is shown in Tab. 1. Many proteins were always highest in serum or in EDTA-plasma, particularly in the DB and GNF sets, respectively. Twenty-four proteins were always lowest in citrate-plasma in the DB set. These specific biases...

The Suitability of Bioremediation

Bioremediation as a biotechnological intervention for cleaning up the residual effects of previous human activities on a site, typically relies on the inherent abilities and characteristics of indigenous bacteria, fungi or plant species. In the present discussion, the emphasis will concentrate on the contribution made by the first two types of organism. The use of plants, including bioaccumulation, phytoextraction, phytostabilisation and rhizofiltration, all of which are sometimes collectively known as phytoremediation, is examined as part of a separate chapter. Thus, the biological mechanisms underlying the relevant processes are biosorption, demethylation, methylation, metal-organic complexation or chelation, ligand degradation or oxidation. Microbes capable of utilising a variety of carbon sources and degrading a number of typical contaminants, to a greater or lesser extent, are commonly found in soils. By enhancing and optimising conditions for them, they can be encouraged to do...

Other natural antimicrobial systems

Antimicrobial compounds, such as sakacin, nisin and reuterin are small proteins produced by gram-positive bacteria and are effective against gram-positive (typically those closely related to the producer strain) and some gram-negative bacterial species (James et al., 1997 Naidu, 2000). In contrast to antibiotics, no residue problems are to be expected (Luecke, 1992). Of particular interest is the anti-listerial activity of nisin and sakacin, and the anti-Salmonella effect of reuterin. In combination with chelating agents, nisin may reduce E. coli 0157 H7 (James et al., 1997), but a single application of nisin on beef inoculated with E. coli 0157 H7 will give a mere 0.4 log10 unit reduction (Cutter and Siragusa, 1995). Nisin is an accepted food additive in various countries (Saltmarsh, 2000). The anti-listerial impact of nisin is also dependent on technology, e.g., high-temperature fermented (US style) sausages (ICMSF, 1998). Nisin as an additive to canned food products aids the...

Neutrophilic Lipopolysaccharide Binding Molecules

Lactoferrin is an 80 kDa glycoprotein that is present in neutrophilic granules, milk, and mucosal secretions (86). Lactoferrin has been shown to bind LPS and to be bac-teriostatic to bacteria, indirectly through chelation of iron ions and directly through destabilization of the gram-negative bacterial cell membrane (86). Lactoferrin is a major LPS-neutralizing compound produced and excreted by stimulated PMN. Lactoferrin peptides containing the LPS-binding site have been shown to prevent the LBP-mediated binding of LPS to CD14, which results in a reduction of TNF and IL-6 release by THP-1 cells in vitro. Germ-free piglets that were fed lactoferrin were less sensitive to LPS, as shown by reduced mortality and hypothermia (87).

Excipientdrug Conjugates

Whether chelation (as opposed to covalent bonding) of an active moiety to an excipient qualifies the entire construct as a new chemical entity (NCE) is a moot point. A case in point is polymer platinate. This compound consists of a polymer backbone, hydroxypropylmethacrylamide, linked to a polypeptide spacer. The peptide is in turn linked with an aminomalonate chelating group, which chelates the platinum compound (Fig. 2). Although it is generally accepted that covalent bonding or chelation of an excipient (eNCE) to a drug (dNCE) seems to qualify the new construct as a NCE, little or no part is played by the eNCE in either the causation or the elicitation of a pharmacological response in the vast majority of drug delivery systems. The authors have therefore classified the eNCE part of the construct as excipient in this discussion. Clinical experience and safety data from such NCEs may serve as supporting evidence to justify the usage of the eNCE as a stand alone excipient in another...

Selditof analysis for protease inhibitor studies

Plasma samples from 20 individuals were collected into identical EDTA-contain-ing tubes with a patented mechanical separator, either with or without the presence of a protease inhibitor cocktail. After collection, samples were processed as quickly as possible (within 15 min) into plasma, hence time zero analyses were from aliquots that were frozen at this earliest time point. Analysis was performed using H50 chips, as per manufacturer's instructions. Briefly, the chips are preincubated with 50 ACN, twice for 5 min each, followed by activation with binding buffer (10 ACN, 0.1 m NaCl, and 0.1 TFA) for 5 min and removed. Additional binding buffer (90 mL) is added, followed by 10 mL of

Solubilization of membrane molecules

The most widely used detergents for membrane solubilization are the nonionic detergents Triton X-100 and NP40 that have almost identical properties. Typically, 1-10 X 107 cells are resuspended in 0.51 ml of the following buffer 50 mM Tris-HCl, 150 mM NaCl, 5 mM EDTA, 10 mM iodoacetamide, pH 8, containing either 0.5-1 Triton X-100 or NP40 and the following protease inhibitors leupep-tin, antipain, chymostatin, pepstatin (all 1 xg ml 1) and 0.1 mM (AEBSF), and lysed at 0 C for 30 min. Insoluble material is subsequently removed by centrifugation at 13 000 g for 15 min.

Preparation from Suspension Cells

Centrifuge again at 400g and discard supernatant. Count cells and resuspend at an appropriate concentration, which will vary with sorter used but will be in the range of 1 X 106-1 X 107 per mL. The final suspension medium will depend on the cell types to be sorted. In general, a low protein concentration is recommended because this will lead to less cell clumping although the addition of 5 mM EDTA (ethylene-diaminetetraacetic acid) will also help this.

Discussion And Summary

The negative effects of phytate in food on human health are likely to be most pronounced in people on marginal subsistence diets that consist mainly of seeds grains fruits. Over two billion humans, largely located in developing countries, have micronutrient deficiencies 42-44 . Considerable scientific literature indicates that the strong chelation capacity of PA can decrease the uptake of elements such as Ca, Fe, and Zn in monogastric digestive tracts 20,45-47 .

Natural Plant Bioremediators

Certain plants, called hyperaccumulators, cope with excess heavy metals in the environment by taking them in and sequestering them in vacuoles, which are bubble-like structures in their cells. Sometimes the plant combines a pollutant with another molecule, a process called chelation. Organic acids often serve this role. Citric acid, for example, surrounds and thereby detoxifies cadmium, and malic acid does the same for zinc. A class of polypeptides called phytochelatins can also bind metals and escort them to vacuoles. Yet a third strategy that plants use to control metal accumulation is to employ a class of small, metal-binding proteins called metal-lothioneins. The intentional use of plants that use any of these ways to take heavy metals from soil is termed phytoremediation. It is a form of bioremediation.

Measurement of Blood Levels

Blood sample collection and processing are critical factors in the determination of homocysteine concentrations. Typically, blood samples for homocys-teine analysis are collected in tubes containing an anticoagulant (e.g., EDTA, heparin). Prompt separation of plasma from the blood cells after centrifuga-tion is required to avoid excess release of intracellular homocysteine into the plasma or removal of homocysteine from the plasma by meta-bolically active leucocytes after blood draw. Keeping the blood sample cold until centrifugation and separation (ideally within 4 h of blood draw) minimizes this problem. Serum homocysteine concentrations typically exceed plasma concentrations by 20 . This is likely due to the fact that blood collected to isolate serum (i.e., without an anticoagulant) must clot at room temperature for 30-60 min before centrifugation and separation. Therefore, plasma is preferred for measurement of homocys-teine. Once separated from the blood cells, the concentration...

Plant Growth Promotion

The plant-growth-promoting capacity of T. harzianum to solubilize in vitro some insoluble or sparingly soluble minerals via three possible mechanisms acidification of the medium, production of chelating metabolites, and redox activity was recently investigated (Altamore et al. 1999). T. harzianum was able to solubilize MnO2, metallic zinc, and rock phosphate (mostly calcium phosphate). Fe2O3, MnO2, Zn, and rock phosphate were also solubilized by cell-free culture filtrates. A size exclusion chromatographic separation of the components of the culture filtrates indicated the presence of a complexed form of Fe but no chelation of Mn. In liquid culture, T. harzianum also produced diffusible metabolites capable of reducing Fe(III) and Cu(II). Solubili-zation of metal oxides by Trichoderma involves both chelation and reduction (Altamore et al. 1999).

Supplementation and Fortification

Fortification of staple foods with 3-10 mg iron daily, depending on the needs of the population, is a long-term preventative strategy. In the United States, bread and cereal products are routinely fortified with 20 mg iron per pound (460 g) of flour, and additional fortification at the option of food suppliers is common. However, fortification is difficult when food processing is decentralized, as is common in poor populations. Food fortification carries the additional challenge that the chemical forms of iron most bioavailable also tend to be the most reactive with the food fortified, resulting in adverse changes in flavor, color, and shelf life. Promising approaches include the fortification of food sauces with iron chemically bound with amino acids or with EDTA (sodium iron ethylenediaminetetraacetic acid), which are well absorbed even in the presence

Factors Affecting Oxidation

The rate of autoxidation in vegetable oils is affected by many factors including degree of unsaturation of fatty acids, metals, and antioxidants. For example, linoleic acid, which contains two double bonds, is oxidized at a faster rate than oleic acid, which contains only one double bond. The relative rates of autoxidation of oleic to linoleic to lin-olenic (3 double bonds) acid has been reported as 1 4050 100 on the basis of oxygen uptake (14). Reasons for differences in autoxidative susceptibility include the fact that the bond strength of linoleic acid (52 kcal mole) is much lower than oleic acid (77 kcal mole), and linoleic acid has a higher reactivity with oxygen than oleic acid. Metals such as copper and iron that exist in two different valence states play an important catalytic role in the oxidation of fats and oils. Metals differ in their ability to catalyze the autoxidation of fats and oils depending on the concentration, reaction temperature, and polarity of the reaction...

Antibodybased measurements of the HUPO reference specimens

We investigated whether the blood preparation methods (serum, citrate-plasma, EDTA-plasma, heparin-plasma) introduced systematic bias into the abundances of all the proteins in general. A systematic bias in concentration would be evidenced by a consistent shift in the concentrations of analytes in one preparation method relative to the other methods. The protein abundances were compared between the samples that were prepared from the same starting material, i.e., we compared the four preparations within the BDAA specimen set, the four preparations within the BDAF specimen set, etc. For each preparation type (citrate-plasma, EDTA-plasma, etc.), the number of proteins that had a maximum concentration in that preparation was totaled. The number of proteins with minimum concentrations also was totaled for each preparation method. Those numbers were compared to the numbers of maxima or minima that would be expected by chance. Frequencies of maxima or minima much greater or lower than would...

Sea Urchin Coelomocytes

In the experiments described below, the sea urchin P. lividus was investigated. The coelomocytes were obtained as a total cell population by bleeding the animals through a cut in the peristomal membrane. The coelomic fluid was poured into an ISO-EDTA anti-coagulant solution. The resulting cell suspension was divided into several Petri dishes. The coelomocytes were harvested in CCM (coelomocytes culture medium) (Henson et al. 1992), kept in Petri dishes, and then exposed to UV or metal stress. Under long-term culture conditions using a modified medium (Le Marrec et al. 1999), the cells tend to form bundles and fibres (see Matranga et al., this Vol., Figs. 4,5).

Effect of Malnutrition on the Course of Tuberculosis

In addition, anemia and hypoalbuminemia are associated with severity of the clinical course of TB. Severe anemia has been negatively correlated with the response to treatment and is associated with early mortality from TB. Anemia in TB patients may be normocytic and normochromic, or it may be microcytic and hypochromic, suggesting a relationship to iron homeostasis in some cases. Iron metabolism is disrupted in active TB. Mycobacterium tuberculosis requires iron to grow and avidly scavenges iron from its environment. One aspect of the host response to TB is intense sequestration of iron that restricts microbial growth. Whether iron supplementation or iron chelation may play a role in the treatment of TB remains speculative.

Endotoxin Nomenclature And Extraction

Endotoxin Structure

Although used interchangeably, Hitchcock and others have proposed reserving the term lipopolysaccharide for purified bacterial extracts, which are reasonably free of detectable contaminants, particularly protein (8) and the term endotoxin for products of extraction procedures, which result in macromolecular complexes of LPS, protein, and phospholipid. Trichloroacetic acid (Boivin-TCA), butanol, and EDTA methods of extractions (8) yield associated outer membrane proteins (OMP) also known as lipid associated proteins (LAP). LAPs, in some cases, have been shown to have potent host effects of their own and act synergistically in association with LPS. Mangan et al. (10) demonstrated that LAP from S. typhimurium is three to four times more active in inducing IL-1 from monocytes than protein-free LPS. Since so many additional components of the bacterial cell wall have recently been implicated in inducing the production of cytokines, this distinction is becoming more widely appreciated....

Mycorrhizae And Bioremediation

The mechanism that confers heavy metal tolerance in mycorrhizal fungi is largely unknown. The survival of AM and EM fungi in polluted soil may depend heavily on the density of the external hyphae. The absorption of heavy metals to the hyphal surface could reduce soil concentrations and thus accumulation of fungal and plant tissue (Denny and Wilkins 1987 Marschner and Dell 1994). Components of the fungal cell wall, such as chitin and melanin, can bind heavy metals to the extraradical mycelium (Denny and Wilkins 1987 Tam 1995). Turnau et al. (1996) found that the EM fungal mantle contained the highest levels of heavy metals while the Hartig net contained the lowest levels. Glomalin, the glycoprotein that coats AM fungal hyphae, could play an equally important role in protecting AM fungi and host plants from toxic metal concentrations in soils, although this has not yet been investigated. Other possible mechanisms conferring heavy metal tolerance in fungi may include intracellular...

Phytatemineral Interactions

Phytate Iron Chelation

The relative binding strengths of different minerals to phytic acid vary greatly. Chelation strength increases with increasing atomic number of the mineral moving from the alkaline earth metals through transition metals in the periodic table. Vohra et al. 13 , using titration curves of phytate as free acid in the presence of single cations, reported that phytate forms complexes with cations in the following descending order of strength

Pcr Inhibition Or Contamination

The use of an internal control is important because impurities or improper nucleic acid concentration may inhibit the PCR reaction, resulting in false negatives. The internal control will give a greater degree of confidence in the validity of a negative amplification result. Every amplification test where inhibitors such as nontarget DNA, heme, bilirubin, bile salts, ethylenediaminetetra-acetic acid (EDTA), sodium dodecyl sulfate (SDS), proteases, and bacterial extracts, etc. are problematic should have an internal control. 5,11,12 To avoid DNA contamination or carried over false positives, the use of deoxyuridine 5'-triphosphate (dUTP) in combination with Uracil-DNA-N-glycosylase (UNG) is recommended. Several other precautions to avoid contamination should be taken, including specific workstations designed for PCR work alone, aseptic techniques, specific pipettes and tips plugged with sterile cotton wool or filter, small aliquots of reagents when performing PCR, and minimizing the...

Importance of trace metals in the culture of aquatic algae

Chelating agents such as citrate and ethylene-diamine tetra-acetic acid (EDTA, usually added as the sodium salt) are also added to the medium. These agents have a high affinity to metal ions, binding with them in a reversible equilibrium where there is always a proportion of free metal ions. The chelating agent (with metal) is too large a molecule to be taken up by the cell, but the presence of free metal ions ensures that there is a continuous low level of availability for metal uptake by algal cells. The addition of these compounds thus provides exploitable soluble reserves of trace metals without problems of toxicity. The chelating agents used in laboratory cultures have the same effect as various organic degradation products (e.g., humic and fulvic acids) and algal secretory products (siderophores) that are present in natural waters.

Morphological Characterization of Yeasts

Solution 1 0.9 M sorbitol and 0.1 M EDTA. Preparation Dilute 1 M EDTA 1 10 and dissolve 163.9 g L of sorbitol. 4. Solution 2 50 mM Tris-HCl and 20 mM EDTA. 9. TE buffer 10 mM Tris-HCl (pH 7.5) and 1 mM EDTA. 3. 1X TAE buffer 40 mM Tris-acetate (pH 8.0) and 1 mM EDTA. Prepare stock solution 50X TAE 242 g L Tris base, 57.1 mL glacial acetic acid, 20 mL of 0.5 M EDTA (pH 7.5). Dilute before use.

Indirect Methods Of Total Lipid Determination

Haugaard and Pettinati 64 have described a turbidimetric method for rapid determination of lipid in milk. The milk fat is homogenized to obtain uniform globules and the milk proteins are retained with chelating agents such as EDTA. Light transmission of the sample is measured and then converted to the lipid content with the use of a conversion chart.

PPP reference specimens

The primary specimens were sets of four reference specimens prepared under the direction of the HUPO PPP Specimens Committee by BD Diagnostics for each of three ethnic groups Caucasian-American (B1), African-American (B2), and Asian-American (B3). Each pool consisted of 400 mL of blood each from one male and one post-menopausal female healthy, fasting donor, collected into 10 mL tubes in a prescribed sequence (see Supplementary Protocol) after informed consent. Very large pools were rejected as requiring too prolonged specimen handling and processing unlike the collection ofindividual specimens even a protocol for two males and two females proved to require more than the 2 h limit we set. Equal numbers of tubes and aliquots were generated with appropriate concentrations of K2-EDTA, lithium heparin, or sodium citrate for plasma or permitted to clot at room temperature for 30 min to yield serum (with micronized silica as clot activator). The additives were dry-sprayed on the inner walls...

Initial Disaggregation And Plating

Gonadal ridges and mesenteries of week 5-9 postfertilization human embryos (obtained as a result of therapeutic termination of pregnancies) are collected in 1-ml ice-cold growth media and rapidly transported to a sterile work space. The tissues are then soaked in calcium-magnesium-free Dul-becco's phosphate-buffered saline (DPBS) for 5 minutes and then transferred to 0.1-ml trypsin-EDTA solution. The concentration of trypsin and EDTA is varied such that at the earliest developmental stages, a gentler 0.05 trypsin-0.5-mM EDTA is used, and at later developmental stages, a stronger 0.25 trypsin-0.5-mM EDTA solution is used. The tissue is mechanically disaggregated thoroughly using a fine forceps and iris scissors. This process is carried out for 5 to 10 minutes at room temperature and then incubated at 37 C for 5 to 10 minutes. This disaggregation process often results in a single-cell suspension and large pieces of undigested tissue. To stop the digestion, serum containing growth media...

Materials and Methods

Dilute 50-500 ng of target DNA to a final volume of 5 mL with 10 mM Tris-HCl (pH 8.5), -0.1 mM EDTA and denature the DNA for 5 min at 98 C in a thermocycler with heated lid. Mix 1.5 mL probe mix (1 fmol of each probe) and 1.5 mL MLPA buffer (1.5 M KCl, 300 mM Tris-HCl pH 8.5, and 1 mM EDTA) and add to the denatured DNA. Heat the mix for 1 min at 95 C before incubating overnight (16 h) at 60 C. After hybridization dilute the sample to 40 mL with ligase mix (2.6 mM MgCl2, 5 mM Tris-HCl pH 8.5, 0.013 nonionic detergents, 0.2 mM NAD) containing 1U Ligase-65 (MRC-Holland) and perform ligation reaction of the annealed probes at 54 C for 15 min. After ligation inactivate the Ligase-65 by incubating the sample 5 min at 98 C. Mix 10 mL of the ligation mix with 30 mL of PCR buffer (20 mM Tris-HCl pH 8.5, 50 mM KCl, 1.6 mM MgCl2, and 0.01 nonionic detergents). While at 60 C, add 10 mL of a solution containing 10 pmol PCR primers, 2.5 nmol dNTPs, and 2.5 U SALSA polymerase (MRC-Holland) to the...

Comparison Of Viral Load Assays

A number of studies have compared the performance of these assays in clinical situations. Schuurman et al. 4 showed that interlaboratory reproducibility of the Roche, Organon, and Versant assays was very good. Others have shown that each of the assays has relatively low intra- and interassay variability.1-5-1 The biological variation of HIV-1 RNA in plasma in clinically stable individuals is approximately threefold (0.5 log) and this variation was not associated with diurnal fluctuations.1-6-1 The anticoagulant used is important for optimal results and the preferred anticoagulant for bDNA is EDTA while citrate or EDTA may be used for NASBA and RT-PCR. Heparin interferes with reagents used in the assays and should be avoided and there is also some question over its effectiveness as a preservative for RNA.

Cytoskeleton Proteins

The partial purification of a 110-kDa peptide, possessing K+EDTA-, Ca2+-, Mg2+-, and F-actin-activated Mg2+-ATPase activities characteristic of myosin-like motor proteins involved in particle vesicle intracellular trafficking, has been reported (D'Andrea et al. 1994). The authors claim that coelomocytes display particle vesicle movements even in cells devoid of microtubules (petaloid) and possess an unconventional myosin which may be the motor protein driving particle vesicle translocation.

Principle of the test

Critical components in the assay are the complement (fresh frozen or lyophilized guinea pig serum) and the sheep red blood cells (El. The latter requires the use of cells from a single or a couple of selected animals. The amount of antibody on E needs to be optimized to a saturating, but subagglutinating, level so that the assay is sensitive to changes in complement activity. The amount of complement is titrated to a level that typically corresponds to twice the amount of the 50 hemolytic dose (CH50) under the test conditions. The amount of complement should be just enough to lyse the indicator cells. Controls for the viability of the detection system (EA + complement), background lysis (no complement) and 100 lysis (lysis by water) need to be included. Reproducibility of the CF assay requires that the ionic strength, pH, Ca2+ and Mg2 content remain constant. Chelators (EDTA, EGTA) should not be used in the test samples because they directly block...

Labeling of DNA Probe by Random Priming

Denature 100-500 ng of satellite DNA e.g., Y chromosome repeats (AATAAAC)n or (AATAC)n or dodeca satellite in water or 10 mM Tris-HCl, 1 mM EDTA, pH 8.0 (TE) buffer by heating at 100 C for 10 min in a 0.5-mL Eppendorf SafeLock microcentrifuge tube. 4. Stop the reaction by adding 2 L of 200 mM EDTA, pH 8.0 (and or by heating at 65 C for 10 min).

Phytate As An Antioxidant In Chicken And Beef

Graf and coworkers 2,12 were the first to evaluate phytate as an antioxidant in a meat system. In cooked, minced chicken breast muscle +10 added water, phytic acid (1.5 mM, pH 6.0) was highly effective in reducing thiobarbituric acid (TBA) values (Figure 12.1) and warmed-over flavor (WOF) intensity (Figure 12.2), compared to controls 12 . Phytic acid (2 mM) was more effective than other antioxidants (2 mM ascorbic acid, BHT, or EDTA) for lowering TBA values in fresh beef homogenates incubated for 60 min at 37 C 13 .

Components Of Liquid And Lyophilized Protein Formulations

A liquid formulation is usually comprised of a buffering agent, a stabilizer (which may also serve as a tonicity agent), a surfactant, and an anti-oxidant when protein oxidation is significant. Chelating agents are employed when metal ion catalyzed reactions predominate. A preservative may be included when a multi-dose formulation is desired.

Surface Treatment and Finish

Passivation is used either after cauterization or as a final treatment of ground, brushed, or polished steels with special surface structures. A natural passive layer is invariably formed when stainless steel is exposed to air. Artificial passivation with dilute nitric acid is frequently used for corrosion resistance at critical condition. The oxidizing effect of nitric acid accelerates the formation of a dense passive layer. Reports on passivation show that organic acid chelating agents can produce a more long lasting passive layer (199).

Materials 21 Cell Culture

4X Hybridization buffer 200 mM HEPES, pH 7.5, 0.8 sodium dodecyl sulfate (SDS), 8 mM EDTA, 2 M NaCl. 9. HE buffer 10 mM HEPES pH 7.5, 1 mM EDTA. 10. Streptavidin solution 2 g pL in 0.15 M NaCl, 10 mM HEPES, 1 mM EDTA, pH 7.4. 12. Phenol saturated with 10 mM Tris-HCl, pH 7.5, 1 mM EDTA. 15. Northern hybridization buffer (100 mL) 1 g BSA, 43 mL H2O, 7 g SDS, 50 mL 1 M sodium phosphate, pH 6.8, 0.2 mL 0.5 M EDTA. 17. RNA gel loading buffer 0.5 mL glycerol, 2 mL 0.5 M EDTA, 5 mg xylene cyanol,5 mg bromphenol blue. 19. Northern washing solution (1 L) 5 g BSA, 50 g SDS, 40 mL 1 M sodium phosphate, pH 6.8, 2 mL 0.5 M EDTA. 20. Northern washing solution II (1 L) 10 g SDS, 40 mL 1 M sodium phosphate, pH 6.8, 2 mL 0.5 M EDTA.

Human Zinc Deficiency

In addition to dietary inadequacy, there are several routes that lead to zinc deficiency. Acrodermatitis enteropathica, the genetic disorder of zinc malabsorption, has already been mentioned. Other, more generalized, malabsorption syndromes (e.g., coeliac disease) can also lead to zinc deficiency. Deficiency has also resulted from inappropriate intravenous feeding and the use of chelation therapy. Children are likely to be particularly at risk of zinc deficiency, because of its involvement in growth.

Modification of the Rapid Mechanistic Plate Assay for HTS

The KSHV Pol-8 PF-8 HTS assay thus employs a reaction buffer containing 50 mM (NH4)2SO4, 20 mM Tris-HCl, pH 7.5, 3 mM MgCl2, 0.1 mM EDTA, 0.5 mM DTT, 2 glycerol, 40 g mL BSA, in addition to 0.625 M dNTPs, 0.125 M DIG-dUTP, and KSHV Pol-8 and PF-8. Z-factors of the modified KSHV Pol-8 Pf-8 assay generally ranged from 0.5 to 0.8.

Protocol 61 Target preparation

Incubate for 30 min at 42 C and add another 1 l SSII for 40 min at 42 C. Add 2.5 l 500 mM EDTA and heat to 65 C for 1min. Add 5 l 1 M NaOH and incubate at 65 C for 15 min to hydrolyze the RNA. Add 12.5 l 1 M Tris immediately to neutralize the pH. Bring the volume to 70 l by adding 35 l of 1 x TE.

Platelet Membrane Fluidity and Other Platelet Tests

Numerous AD-associated phenomena have been described in platelets (Table 8). An increase in the fluidity of platelet membranes in persons with early-onset and late-onset AD using fluorescence spectroscopy has been most extensively studied, and furthermore has been evaluated in a longitudinal study (427). To study platelet membrane fluidity, purified platelets obtained from fasting blood samples are collected in the morning in a plastic syringe containing EDTA as an anticoagulant and a protease inhibitor are labeled with the lipid probe 1,6-diphenyl-1,3,5-hexatriene (DPH), and the steady-state anisotropy of the DPH labeled membranes is determined at 37 C. An increase in platelet membrane fluidity is reflected by a decrease in the steady-state anisotropy of the labeled membranes. In 1987, 71 of patients were reported to have lower anisotropy values than 8 of the controls. The alteration in platelet fluidity parallels clinical severity as measured by the Mini-Mental State Exam (MMSE)...

Nebulizers and Aqueous Based Systems

Aqueous formulations for nebulization generally follow the same principles as those adopted for the development of parenteral products. Excipient selection is based upon achieving pH, osmolarity, and sterility suitable for deposition in the lung. In addition, the stability of formulations, whether solution or suspension based, follows typical guidelines adopted for any aqueous delivery system. Isotonicity is typically achieved by adding buffer salts or sodium chloride (9). Preservatives such as EDTA and benzalkonium chloride have been included in many formulations, but there has been some concern over the possible physiological implication of acute and chronic exposure to these compounds (9). Thus, to manufacture sterile aqueous-based oral inhalation solutions and suspensions, the unit-dose production and packaging in sealed nebules or similar packaging is recommended to prevent microbial contamination. The use of preservatives or stabilizing agents in inhalation spray formulations is...

Lysis Buffers And Other Solutions

Ature. 44,55,56 The various formulations for ''lysis buffers'' generally consist of a Tris buffer at a pH between 7.5 and 8.0, EDTA (chelator), and sodium dodecyl sulfate or n-lauroylsarcosine (detergents to lysis cells). These buffers have demonstrated to be effective at room temperature in the preservation of DNA in blood samples for 6 months 44 and tissue samples for 2 years or more. 39 The use of a dimethyl sulfoxyde (DMSO) salt solution for the preservation of DNA in various tissues has been demonstrated to be effective for storage from 6 months to over 2 years. 39,44,48 This solution of 20 DMSO, 0.25 M EDTA, and NaCl 5 to saturation, pH 7.5 44,48 was found to be the most effective method of noncryogenic storage for the prevention of DNA degradation. 39


Lactoferricin B or hydrolyzed lactoferrin (HLF) is a small peptide produced by acid-pepsin hydrolysis of bovine lactoferrin (Bellamy et al., 1992). Jones et al. (1994) reported that the compound was inhibitory to Shigella, Salmonella, Yersinia enterocolitica, E. coli 0157 H7, S. aureus, L. monocytogenes and Candida. In contrast, while HLF was effective against L. monocytogenes, Enterohemorrhagic E. coli, and Salmonella Enteritidis in peptone yeast extract glucose broth, it was not active in a more complex medium, trypticase soy broth (TSB) (Branen and Davidson, 2000). The addition of EDTA enhanced the activity of HLF in TSB, indicating that the decreased activity of HLF may have been due, in part, to excess cations in the medium. Venkitanarayanan et al. (1999) found that, while 50 or 100 pg lactoferricin B per ml reduced viable E. coli 0157 H7 in 1 peptone, it was much less effective as an antimicrobial in ground beef.

Natural resistance

The dog, like most multicellular organisms, has a vast array of defense mechanisms. Some of the most effective defense mechanisms are not part of the immune system but are classified as natural resistance. Natural resistance is defined as defense mechanisms which prevent or combat disease-causing agents by nonimmunologic means (without memory or specificity). Examples of canine natural resistance are barriers (skin, mucous membranes), commensal organisms (normal bacterial and viral flora), mechanical action (mucociliary tree, coughing, and desquamation), flushing action (urine, saliva, tears, milk and diarrhea), chemical and enzymatic action (gastric acid, digestive enzymes, lysozyme, and complement), nonspecific phagocytosis (macrophages and polymorphonuclear neutrophils), hormonal (a and 3 interferon), iron-chelating agents (lactoferrin), and natural killer cells. Although the dog does not have unique natural resistance mechanisms, it is important to understand the role of natural...


In patients with other forms of secondary dystonia, a careful history and physical and neurologic examination are essential to investigate for the underlying cause. An important secondary dystonia to consider is Wilson's disease. To assess for this disease, a slit lamp examination for Kayser-Fleischer rings, a serum ceru-loplasmin, and a 24-hour urine test for copper are recommended. A patient with Wilson's disease may be treated successfully by chelation therapy.


It is present in avian eggs, mammalian milk, tears (and other secretions), insects and fish. In hypotonic solutions, the enzyme causes lysis of bacterial cells. The enzyme is most active against gram-positive bacteria because the peptidoglycan of the cell wall is more exposed. It inhibits the foodborne bacteria Bacillus stearothermophilus, Clostridium botulinum, C. thermosaccharolyticum (Thermoanaerobacterium thermosaccharolyticum), C. tyrobutyricum, Listeria monocytogenes and Staphylococcus aureus (Hughey and Johnson, 1987 Hughey et al., 1989). There is variation in the susceptibility of gram-positive bacteria to lysozyme probably due to the presence of teichoic acids or other compounds that bind the enzyme. Also, certain species have greater proportions of 1,6 or 1,3 glycosidic linkages in the peptidoglycan which are more resistant than the 1,4 linkage (Tranter, 1994). Lysozyme is less effective against gram-negative bacteria due to reduced peptidoglycan...

Dr Eric W Deutsch

HUPO initiated the Plasma Proteome Project (PPP) in 2002. Its pilot phase has (1) evaluated advantages and limitations of many depletion, fractionation, and MS technology platforms (2) compared PPP reference specimens of human serum and EDTA, heparin, and citrate-anti-coagulated plasma and (3) created a publicly-available knowledge base pride). Thirty-five participating laboratories in 13 countries submitted datasets. Working groups addressed (a) specimen stability and protein concentrations (b) protein identifications from 18 MS MS datasets (c) independent analyses from raw MS-MS spectra (d) search engine performance, subproteome analyses, and biological insights (e) antibody arrays and (f) direct MS SELDI analyses. MS-MS datasets had 15 710 different International Protein Index (IPI) protein IDs our integration algorithm applied to multiple matches of peptide sequences yielded 9504 IPI proteins identified with one or more peptides and 3020 proteins identified with two...


Thallium is a potent rodenticide that has largely fallen out of favour since the 1980s owing to its toxicity. The use of this agent is banned in some countries. Its mode of action involves interference with nerve transmission and mitochondrial function. Clinical signs of toxicity are either acute or chronic in nature. The acute form is characterised by vomiting, depression and marked abdominal pain, whereas chronic cases are manifested by neurological deficits and severe skin and mucosal changes, such as erythema, alopecia and ulceration. The prognosis is usually poor in such cases, with treatment being principally directed at minimising any further gastrointestinal tract absorption using Prussian blue or activated charcoal. Potassium chloride has been used to accelerate renal excretion of thallium. The use of chelating agents, such as diphenylthiocarbazone, is controversial, and generally no longer recommended.

Snow Crab

Cause(s) for the discoloration (12), it may be the result of the enzymatically mediated oxidation and polymerization of phenolic compounds naturally present in crab (2). Thus antioxidants, sulfites, and chelating agents have been used to block the bluing discoloration. However, speed or quick processing is the best assurance in preventing the bluing, which will still occur and even develop at a faster rate after the frozen raw crab section is thawed. If crab are not cooked properly to destroy these enzymes, then the crab should be further processed quickly or consumed soon after thawing.