Ethylenediaminotetraacetic Acid Research Articles

Role of Ca++‐Dependent Proteases and Lysosomal Enyzmes in Postmortem Changes in Bovine Skeletal Muscle

ABSTRACTBovine longissimus muscles excised at slaughter were compared to cross‐sectional slices of contralateral longissimus muscles obtained 12 hr postmortem for calcium‐dependent protease (CDP), cathepsins B, H and L activity, and myofibrillar fragmentation index (MFI). Slices were suspended in either Tris‐acetate buffer, buffer + ethylene diaminotetraacetic acid (EDTA), buffer + ethylene glycol‐bis (β, aminoethyl ether) N, N, N', N'‐tetraacetic acid (EGTA) or buffer + CaCl2 for either 1, 3, or 7 days while the postmortem associated changes were followed. EDTA, EGTA and Ca++ had no effect on cathepsin B, H or L activities. EDTA and EGTA treated slices had 149% whereas Ca++ 48% of control CDP activity. Postmortem changes were completed after 24 hr of Ca++ treatment but did not occur in EDTA‐ and EGTA‐treated slices. Thus, the changes observed during postmortem storage appeared to be associated with CDP activity rather than catheptic enzymes.

Phase transformation of titanium dioxide in the preparation of titanium oxide-Vanadium oxide electrodes for complexometric titration

Vanadium(V) oxide and vanadium-titanium oxides supported on titanium were examined by electrochemical measurement, Raman spectroscopy, scanning electron microscopy and X-ray diffraction to assess their use for amperometric end point indication of complexometric titrations. The electrodes were prepared by coating titanium sheets or wires with mixed solutions of vanadium(V) and titanium(IV) and heating. The decomposition is accompanied by a phase transformation of the titanium dioxide from the anatase to the rutile lattice. The electrode has high activity for the electro-oxidation of ethylenediaminotetraacetic acid (EDTA) in alkaline buffered solution. It also performs better than other electrodes made by anodic deposition of transition metal oxides on platinum.

Partial characterization of α‐mannosidase from Yarrowia lipolytica

AbstractAn exo‐α‐mannosidase (E.C. 3.2.1.24) was characterized in the yeast form of Yarrowia lipolytica. The enzyme located in a crude particulate fraction of the cell extract is under catabolite repression, has an optimum pH of 6.0, a Km of 0.27 mM with p‐nitrophenyl‐α‐D‐mannopyranoside and is partially inhibited by D‐mannose.The enzyme is not affected by ethylenediaminotetraacetic acid, several cations (only Zn++ increased its activity in a 25%) or sulphydryl reagents and can be partially solubilized by treatment with digitonine.

Pod Retention and Seed Yield of Beans in Response to Chemical Foliar Applications

Abstract Chemicals often associated with pollen function in vitro were applied under field conditions to foliage of determinate, semi-determinate, and indeterminate beans (Phaseolus vulgaris L.) to evaluate pod and seed yield response. Sprays of calcium nitrate, boric acid, ethylenediaminotetraacetic acid, detergent “Micro”, and different sugars altered pod retention and seed yield, but response varied with bean source.

A novel IgA protease from Clostridium sp. capable of cleaving IgA1 and IgA2 A2m(1) but not IgA2 A2m(2) allotype paraproteins.

Three bacterial strains of Bifidobacterium and Clostridium sp. from patients with inflammatory bowel disease (I.B.D.) and Streptococcus pneumoniae from a patient with pneumonia were identified to produce extracellular proteases cleaving IgA into Fab and Fc fragments. Although the proteases from the Bifidobacterium and the Streptococcus pneumoniae showed the characteristics of typical IgA1 proteases, cleaving the IgA of only the IgA1 subclass, the protease from Clostridium sp. revealed a dual substrate specificity, in that it cleaved both IgA1 and IgA2 of the A2m(1) allotype. The latter protease, however, did not show any activity with respect to the IgA2 of the A2m(2) allotype. Fc fragments isolated from the IgA1 and the IgA2 A2m(1) by digestion with the Clostridium sp. protease were identified to have an identical amino terminal residue of valine. The site of cleavage in both the alpha 1 and the alpha 2 of A2m(1) by the protease was assumed to be an identical peptide bond at Pro(221)-Val(222), which is a common one present just before the hinge of both the alpha 1 and the alpha 2 of the A2m(1) but not of the alpha 2 of the A2m(2). The protease was sensitive to ethylene-diamino tetraacetic acid, a chelating agent, similar to other already reported IgA1 proteases.

Growth hormone secretion in chicks during dietary calcium deficiency

The role of calcium in the secretion of growth hormone (GH) has been examined in vivo in immature domestic fowl. Chicks reared on a low calcium (0.16% calcium in the feed) diet showed a reduced growth rate, compared with those on a normal (0.86%) calcium diet and had lower basal levels of plasma GH 1, 5, 10 and 15 d after calcium deprivation, but not after 20, 25, 30 or 35 d of calcium deficiency. No consistent changes in the concentration of immunoreactive somatomedin C were observed during calcium restriction. In both the control and low calcium fed birds the plasma concentrations of GH were elevated by the intravenous administration of human pancreatic GH releasing factor (hpGRF) and by thyrotropin releasing hormone (TRH). The GH responses to these provocative stimuli were not reduced in magnitude by calcium deficiency. It is suggested, therefore, that the effect of calcium deprivation on the secretion of GH is mediated via the reduced release of stimulatory hypophysiotrophic factors from the hypothalamus. Pharmacological alterations in calcium status also suggest that calcium deprivation inhibits GH secretion. Plasma concentrations of GH were acutely depressed in young chicks following the administration of a calcium chelator (ethylene diaminotetraacetic acid) a calcium channel blocker (verapamil) and after calmodulin inhibition (by chlordiazapine and trifluorpenazine administration). These data therefore demonstrate the importance of calcium in the stimulation of GH secretion in the domestic fowl.

Role of calcium and the calcium-calmodulin complex in resumption of meiosis, cumulus expansion, viability and hyaluronidase sensitivity of bovine cumulus-oocyte complexes.

The necessity of calcium (Ca2+) and the Ca2+-calmodulin complex for resumption and completion of meiosis, expansion of cumulus cells, viability and hyaluronidase sensitivity of in vitro cultured bovine cumulus-oocyte complexes was examined by inhibition of the Ca2+-calmodulin complex with eight graduated doses of trifluoperazine (TFP) and by Ca2+ deficiency or depletion. Doses of TFP greater than 2.5 microM decreased the percent of cumulus complexes surviving culture and oocytes completing meiosis, whereas cumulus expansion was unaffected until the cultures contained a near lethal dose (greater than 10 microM). Hyaluronidase caused dispersion of cumulus cells whenever they were expanded regardless of TFP dose. In TC-199 media the completion of meiosis I was suppressed by 0.1 to 1 mM ethylenediaminotetraacetic acid (EDTA) (P less than 0.05) and drastically reduced by 1.0 mM (P less than 0.05). Viability of the cumulus-oocyte complex was not reduced until the dose of EDTA was increased to 1.0 mM (P less than 0.0001). Cumulus expansion was also not suppressed until the dose of EDTA reached 1.0 mM (P less than 0.05). In Ca2+-free (CF) basal media Eagles, completion of meiosis I was reduced by all doses of EDTA (P less than 0.05), whereas viability of the cumulus-oocyte complex was decreased by Ca2+ deficiency or by EDTA addition to basal media Eagles (P less than 0.01). Cumulus expansion was unaffected by Ca2+ removal or chelation. In all experiments, oocytes which were not degenerate underwent germinal vesicle breakdown regardless of treatment.

β 2-glycoprotein-I (apolipoprotein H) interactions with phospholipid vesicles

1. 1. The binding characteristics of the human serum protein β 2-glycoprotein-I, also called apolipoprotein H, with multilamellar phospholipid vesicles has been studied. 2. 2. It was found that β 2-G-I is not or almost not bound to the “neutral” phospholipids phosphatidylcholine (PC), phosphatidylethanolamine (PE) and sphingomyelin (SM). 3. 3. The negatively charged compounds phosphatidylserine (PS) and phosphatidylinositol (PI) interact strongly with β 2-G-I. In terms of phospholipid concentration the binding to PS is about one order of magnitude greater than to PI. 4. 4. The binding capacity is influenced by several parameters such as the molarity of buffer, presence of mono- or divalent cations as well as ethylenediaminotetraacetic acid (EDTA). Proteins like bovine serum albumin (BSA), human serum albumin (HSA) or horse γ-globulin (HGG) influence the binding also in a concentration dependent manner.

Regulation of androgenesis inNicotiana tabacum L. cv. White Burley andDatura innoxia Mill. Effect of bivalent and trivalent iron and chelating substances

The effect of FeSO4.7H2O, Fe2(SO4)3.9H2O, disodium salt of ethylene-diaminotetraacetic acid, dihydrate (EDTA) and N-(2-acetamido) iminodiacetic acid (ADA) and their combinations on the androgenesis was studiedin vitro in tobacco (cv. White Burley) and datura (Datura innoxia Mill.). Simultaneously the reversibility and irreversibility of the morphogenic process leading to the conversion of the pollen embryoid into complete plant was followed. Complete plants developed in anthers on media with trivalent iron, chelated trivalent iron, chelated bivalent iron, bivalent iron in the presence of ADA and of media with EDTA. The number of androgenic plants in anthers increased in the following order: Fe3+ < Fe3+ EDTA ≦ ≦ EDTA < Fe2+ EDTA. The marked brown colour of cultured anthers was due to the presence of trivalent iron in the medium. The androgenic development was most rapid on the medium containing only trivalent iron, slower on media with chelated iron and slowest on medium with EDTA. The viability of cultures with complete plants decreased in the reverse order. No complete plants grew on media without trivalent iron and without EDTA and on media containing only bivalent iron whereas globular embryoids arose and developed continuously on these media. The anthers reacted in the same way on both complete and minimal media. Isolated embryoids formed complete plants in corresponding variants on complete media only. The development of pollen embryoids into complete plants was stopped by the transfer of globular and torpedo-shaped embryoids from medium with EDTA to the medium without EDTA. Isolated greenish cotyledonar embryoids continued to grow even on the medium without EDTA.

Effect of aluminosilicate composition on acid and catalytic properties of dealuminized zeolites

1. When various samples of synthetic faujasite are dealuminated by (ethylenediamino)tetraacetic acid to the same Si/Al ratio the number of aluminum atoms in the unit cell differs and depends on the aluminosilicate composition of the starting sample. 2. At a constant Si/Al ratio the samples with a smaller number of aluminum atoms in the unit cell have a greater adsorption capacity toward ammonia and a higher catalytic activity. 3. The properties of zeolites with a faujasite structure depend not only on the Si/Al ratio, but also on the absolute number of aluminum atoms in the unit cell.

A Fine Structural Study of Mineralization Foci in Antler and Growth Plate Cartilage

Established staining techniques for glycosaminoglycans (GAG) and established demineralization techniques were employed to investigate the possible relationship of GAG's to hydroxyapatite crystals in mineralized cartilage. Cervine antler cartilage and rat growth plate cartilage were subjected to glutaraldehyde (6.25%) fixation, demineralized with ethylene diaminotetra acetic acid (EDTA) or acidic cacodylate buffer (pH 5.5), stained and postfixed with a ruthenium red (RR)-osmium tetroxide solution and prepared for routine transmission electron microscopy. Some tissues were subjected to hyaluronidase digestion prior to RR staining. Sections were stained with RR only or RR and uranyl acetate. The sections were examined with a Hitachi HS-8 electron microscope.Matrix granules were positive to RR staining (Fig. 2&amp;3). The granular density of the interterritorial zone was higher than that of the lacunar area; however, the lacunar granules were larger. After EDTA treatment, mineralization foci in the interterritorial zone were devoid of hydroxyapatite crystals.

Adrenal cortex adenylate cyclase. Specific binding sites for 5'-guanylyl-imidodiphosphate in partially purified plasma membranes from bovine adrenal cortex.

Specific binding sites for 5'-guanylyl-imidodiphosphate [Gpp(NG)p] have been identified in a partially purified plasma membrane fraction from bovine adrenal cortex. The apparent affinity of Gpp(nh)p at 30 degrees C was 12muM-1 and the concentration of binding sites was 100 pmoles per mg of protein. Binding of Gpp(NH)p is inhibited by Mn2+ greater than Mg2+ greater than Ca2+ and enhanced by low concentrations of the chelators ethylenediamino-tetraacetic acid (EDTA) and ethylene glycolbis-(beta-aminoethylether))-N,N'-tetraacetic acid (EGTA). High concentrations of EDTA are inhibitory and at 2.5 mM EDTA binding of Gpp(NH)p is only 10% of that observed in the absence of the chelator. The bound labeled GTP analogue exchanged only slowly with the unlabeled nucleotide after a steady state has been reached. EDTA also releases the bound labeled Gpp(NH)p from its binding sites. The slow dissociation of Gpp(NH)p can explain the persistent activation of adenylate cyclase observed after pretreatment of bovine adrenal cortex plasma membranes with Gpp(NH)p and subsequent washing. It is suggested that at least parts of these binding sites are identical to the sites identified earlier as regulatory sites for angiotensin high-affinity receptors (Glossmann et. al., 1974a) and for ACTH-stimulated cyclase Glossmann and Gips, 1974).

Schistosoma mansoni: Mechanism of cercarial tail loss and its significance to host penetration

The mechanism of tail loss from cercariae of Schistosoma mansoni was investigated under experimental conditions. Tail loss from 90% or more of cercariae occurred in less than 10 min when packed organisms were incubated at 30 C in a minimal volume of water. Proteolytic secretions from the acetabular glands did not play a significant part in this process since the addition of known protease inhibitors nor the addition of secretions collected from other cercariae influenced the rate of the process. Tail loss was inhibited when the packed cercariae were incubated in saline at concentrations of 0.05 M or above though glandular secretion occurred at an equal rate in both water and saline. Tail loss was also inhibited by the chelating agents ethylenediaminotetraacetic acid (EDTA) and ethyleneglycoltetraacetic acid (EGTA). A marked feature of cercariae packed in water was their clumping and adhesion to the walls of the containing vessel by secretions from the postacetabular glands. Such clumping did not occur in saline or chelant solutions. It is suggested that the most probable mechanism for tail loss is a simple mechanical trauma effected by the movement of the tail acting against the resistance of the secretion-fixed body. During incubation under conditions which remove the “coat” from the body of the cercaria, the “coat” of the tail remains intact.

Bacteriocinogenic Activity in the Genus Cellvibrio

SUMMARY: The production and characteristics of a new bacteriocin, cellvibriocin, are described. Of ten known strains of Cellvibrio examined six were cellvibriocino-genic. Cellvibriocin produced by Cellvibrio sp. 9916 appeared most specific on most media and was studied in detail. It was thermolabile, unaffected by deoxy-ribonuclease, ribonuclease, lysozyme and catalase but completely inactivated by proteases and protein denaturants such as chloroform, acetone and ethanol. Although not readily detectable nor inducible in liquid culture cellvibriocin was induced on solid (agar) media by ultraviolet light; it was not extractable from agar grown cultures by buffer solutions or by freeze-thawing, or from liquid grown cultures lysed with ethylene diaminotetraacetic acid and lysozyme or from cultures disintegrated by ultrasonication, in a Hughes&apos;s press, with butanol or with alumina.

A technique for the fixation and decalcification of rat incisors for electron microscopy.

A reproducible technique that provides sections of decalcified rat incisors for electron microscopy is described. Fixation is accomplished by perfusion with slightly hypertonic, neutral phosphate-buffered 2.5% glutaraldehyde for 30-60 min. Throughout the rest of the procedure, isotonic neutral solutions are used. For decalcification, 4.13% disodium ethylene diamino tetraacetic acid for 14-21 days is employed, followed by washing in phosphate buffer for 2 days, postfixation in 1% osmium tetroxide for 4 hr and embedding in Epon. Provided that the initial fixation by perfusion is good, the method gives satisfactory preservation of the fine structures of the cells and matrices of the incisor, in addition to maintaining the relationship of hard and soft tissues.

New methods of photometric determination of microgram quantities of ethylenediamino tetraacetic acid

New methods of photometric determination of microgram quantities of ethylenediamino tetraacetic acid

HEPATIC AND RENAL STUDIES WITH IRON-59 EDTA IN PATIENTS WITH AND WITHOUT LIVER OF KIDNEY DISEASE.

The use of ethylenediaminotetraacetic acid (EDTA) is not new to the field of radiology. Sapeika (4–6) and Shapiro (7) attempted to use the lead chelate of this compound for contrast radiology during the midportion of the last decade. Excellent opacification of the gastrointestinal tract, vascular structures, and the urinary tract was obtained. No toxic effects were noted following oral administration, but Shapiro reported increased irritability and generalized convulsions, promptly followed by respiratory failure and death, when doses of 800 mg. per kilogram were administered intravenously to rabbits. Two suggestions were offered to explain the deaths on the basis of possible acute lead poisoning. One was the equilibration of the undissociated chelated lead in vivo with the various physiological cations, resulting in the release of free ionic lead. The second was the cleavage of the lead chelate in the liver, with liberation of ionic lead. Because of this latter possibility, and since iron 59 possessed fa...

Kinetics of electrode processes of complexes in polarography. VII. Formation of the complex of cadmium ion with the ethylenediaminotetraacetic acid as a reaction deactivating the product of a rapid electrode reaction

Kinetics of electrode processes of complexes in polarography. VII. Formation of the complex of cadmium ion with the ethylenediaminotetraacetic acid as a reaction deactivating the product of a rapid electrode reaction

The simultaneous determination of copper and iron in high purity aluminium using the k 1000 cathode ray polarograph

An accurate simultaneous determination of copper and iron in 99 99% aluminium can be carried out in 10–15 min, using the linear sweep polarograph. Lead can also be determined in the same solution The basic electrolyte used is sodium hydroxide and sucrose, but the addition of ethylene-diaminotetraacetic acid (EDTA) is necessary if manganese is present. The effect of EDTA on the reduction wave of copper and iron has been investigated

A proteolytic system in growing and non-growing cells ofEscherichia coli

(1) Escherichia coli cells contain a protease system splitting its own proteins as well as casein labelled with131I at a slightly alkaline pH. The rate of proteolysis is not markedly influenced by ethylenediaminotetraacetic acid or by iodoacetic acid, p-chlormercuribenzoate diminishes the rate of hydrolysis of131I casein but its effect cannot be reversed by cysteine. (2) The proteolytic system is not present in the cells in the form of an enzymogen and its activity is not diminished even after 55 passages on a synthetic medium containing ammonium chloride as the sole nitrogen source. (3) In living cells proteins labelled with36S methionine are degraded at a rate not exceeding 0.2%/hr. at 29–30°, both in a growing culture and in the normal stationary phase. Similarly, the protease activity in cell-free extracts does not fundamentally change in the course of development of the culture. (4) Proteins in extracts are split at a rate of 1.8–2.3%/hr. at 32° according to results based on colorimetric estimations with tyrosine, or at a rate of 1.1–1.3%/hr, at 32° as shown by the course of degradation of35S proteins. No resynthesis of proteins from liberated amino acids took place in the extracts under the described experimental conditions. (5) The decomposition of labelled proteins in cell-free systems was stimulated by dinitrophenol and sodium azide.