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Influence of EDTA and citrate on physicochemical properties of whey protein-stabilized oil-in-water emulsions containing CaCl2.

The influence of chelating agents (disodium ethylenediaminetetraacetate (EDTA) and sodium citrate) on the physicochemical properties of whey protein isolate (WPI)-stabilized oil-in-water emulsions containing calcium chloride was determined. The calcium-binding characteristics of EDTA and citrate at 30 degrees C were characterized in aqueous solutions (20 mM Tris buffer, pH 7.0) by isothermal titration calorimetry (ITC). EDTA and citrate both bound calcium ions in a 1:1 ratio, but EDTA had a much higher binding constant. Oil-in-water emulsions (pH 7.0) were prepared containing 6.94% (w/v) soybean oil, 0.35% (w/v) WPI, 0.02% (w/v) sodium azide, 20 mM Tris buffer, 10 mM CaCl(2), and 0-40 mM chelating agent. The particle size, apparent viscosity, creaming stability, free calcium concentration, and particle surface potential of the emulsions were measured. The chelating agents reduced or prevented droplet aggregation in the emulsions. When they were present above a certain concentration (>3.5 mM EDTA or >5 mM citrate), droplet aggregation was prevented. The reduction of aggregation was indicated by decreases in particle size, shear-thinning behavior, apparent viscosity, and creaming. Emulsions containing chelating agents had lower free calcium concentrations and more negatively charged droplets, indicating that the chelating agents improved emulsion stability by binding calcium ions. EDTA could be used at lower concentrations than citrate because of its higher calcium ion binding constant.

Calcium Chloride↗

Ultrastructural studies on mitochondrial swelling.

Mitochondrial swelling induced by valinomycin, calcium chloride and P(i) was studied after potassium permanganate fixation in suspension. Valinomycin induces a rapid K(+) influx, increase of the matrix space and out-folding of the cristae, with good preservation of the matrix material. This swelling is reversible but the cristae do not completely re-form and have a blebbed appearance. On repeated swelling and contraction cycles there is a gradual loss of matrix material. Calcium chloride and P(i) produce a slow swelling of the matrix space. Shrinkage induced by ATP was partial and not associated with return to the original structure.

Adenosine Triphosphate↗

Quantification of growth factor levels using a simplified method of platelet-rich plasma gel preparation.

PURPOSE: This study compared two methods of preparing platelet-rich plasma (PRP) gel and the levels of PDGF and TGFbeta in each preparation. MATERIALS AND METHODS: Platelet-rich plasma gel was prepared by centrifugation and clotted using the ITA gelling agent (Natrex Technologies Inc, Greenville, NC) or by the addition of thrombin and calcium chloride. The levels of platelet-derived growth factor (PDGF) and transforming growth factor beta (TGFbeta) generated by clot formation were assayed by enzyme-linked immunoassay (ELISA). RESULTS: Both methods of preparation yielded PRP gel in less than 30 minutes. However, the ITA preparation did not require thrombin to achieve adequate gel formation. The levels of PDGF and TGFbeta were similar regardless of which method was used for initiation of clot formation. CONCLUSION: Use of ITA for gel preparation is equivalent to using calcium chloride and thrombin, without the need for special equipment and the risk of coagulopathy.

Blood Coagulation↗

Calcium-cadmium interaction on L-threonine intestinal transport.

Cadmium is a highly toxic metal that can damage a number of organs including the gastrointestinal tract. It has been shown that cadmium partially reduces L-threonine intestinal absorption probably by binding to membrane proteins which pertain to active transport systems or are functionally related to them. Calcium, however, is an essential element in a wide variety of cellular activities. The aim of the present work was to study whether the inhibitory cadmium effect on L-threonine absorption across rabbit jejunum could be modified by calcium. In media with Ca2+, cadmium significantly reduces the L-threonine absorption. In Ca(2+)-free media, where calcium chloride was omitted and replaced isotonically with choline chloride, this amino acid transport was not modified by cadmium but it was inhibited when calcium chloride was replaced isotonically with magnesium chloride. Verapamil (blocking mainly Ca2+ transport) did not modify the inhibitory effect of cadmium on L-threonine transport. When A 23187 (Ca2+ specific ionophore) was added in media with/without Ca2+, cadmium produced no change on L-threonine transport. These results suggest that calcium and cadmium could have an affinity for the same chemical groups on the enterocyte membrane. This property could affect the intestinal absorption of amino acids.

Animals↗

On the solubility of calcium deoxycholate: kinetics of precipitation and the effect of conjugated bile salts and lecithin.

In view of the low solubility of calcium deoxycholate and the possible induction of cholesterol precipitation in the gallbladder by calcium insoluble salts, we find it of interest to study the precipitation of calcium deoxycholate and its dependence on other bile components. The findings of these studies were as follows: (i) Precipitation of calcium deoxycholate from mixtures of calcium chloride and monomeric deoxycholate (at concentrations below the critical micelle concentration (CMC] is very slow even at relatively high CaCl2 concentrations (more than 20 days at 50 mM CaCl2). (ii) At higher deoxycholic acid (DOC) concentrations, precipitation of micellar DOC is faster and requires much lower calcium chloride concentrations. For any given calcium concentration, the rate of precipitation is maximal at an optimal DOC concentration. In solutions containing 150 mM NaCl, the maximal rate of precipitation occurs at about 10 mM DOC, almost independent of Ca2+ concentration. At lower ionic strength (10 mM NaCl), the optimal DOC concentration is 30 mM. These observations suggest that the most important factors in determining the rate of Ca(DOC)2 precipitation are (a) the ratio between calcium ions bound to the surface of a DOC micelle, and the [DOC] (the Ca2+/DOC binding ratio) and (b) the concentration of DOC micelles. (iii) In the presence of conjugated deoxycholates, the crystallization of calcium deoxycholate is inhibited. Phosphatidylcholine has a similar, although smaller, inhibitory effect. Upon precipitation of calcium deoxycholate from a mixed micellar system containing sodium deoxycholate, phosphatidylcholine and cholesterol, the latter two components spontaneously form vesicles. The anti-nucleating effect of PC and conjugated bile salts is explained in terms of "poisoning" of the crystallization process. In view of the latter results we conclude that under normal conditions calcium deoxycholate is not likely to precipitate in the gallbladder.

Bile Acids and Salts↗

[The dose-dependent effects of a combination of different classes of antioxidants exemplified by dibunol and beta-carotene].

In order to enhance antioxidant protection of the organism in various pathological states, effective combinations of antioxidants have been developed that interact with various types of free radicals. On two experimental models,--acute alcohol intoxication and calcium chloride arrhythmia of rats, the prophylactic activity of antioxidant combinations was established for certain doses, rather then for monotherapy. The possibility of obtaining high protective effects at relatively low concentrations of the components in the complex preparation was shown. Am increase of antioxidant concentration in the preparation could decrease its efficiency. The highest efficiency in the complex preparations, as compared with in each component taken separately, was observed in the case of calcium chloride arrhythmia when dibunol and beta-carotene were combined at concentrations of 10 mg/kg and in the case of acute alcohol intoxication (1 and 0.25 mg/kg, respectively). Enhanced efficiency of combined antioxidant therapy opens up the way to the production of new active complex preparations with minimum toxic side toxic effects and complications.

Alcoholic Intoxication↗

Comparative toxicity of a zinc salt, zinc powder and zinc oxide to Eisenia fetida, Enchytraeus albidus and Folsomia candida.

The pore water zinc concentration and the calcium chloride extracted zinc fraction are higher in the soils spiked with a zinc salt (ZnCl2) compared to soils spiked with zinc oxide or zinc powder. Based on total zinc concentrations in the soil, the acute toxicity of zinc salt to the compost worm Eisenia fetida, the potworm Enchytraeus albidus and the springtail Folsomia candida was lower compared to zinc oxide and zinc powder. However, when expressed on the basis of pore water concentrations or calcium chloride extracted fractions, acute toxicity was higher for zinc salt, which indicated that dermal uptake via the pore water is not the only route of uptake. Chronic toxicity of zinc salt, zinc oxide and zinc powder was similar when based on total concentrations in the soil which again indicates that the pore water route of uptake is not the only route of exposure but that oral uptake is also important.

Animals↗

In vivo continuous monitoring of ionized calcium in dogs using ion sensitive field effect transistors.

The authors have used calcium ion (Ca++) sensitive field effect transistor (ISFET) probes to monitor in vivo changes in Ca++ activity in the venous blood of dogs in response to infusions of sodium citrate or calcium chloride. During eight experiments, Ca++ activity changes were monitored in response to sodium citrate infusions simulating infusion of 1, 2, or 4 units of citrate preserved blood per 6 min. The ISFET probe recorded decreases in Ca++ activity during all infusions. Periodic serum samples were analyzed off-line using an in vitro ion selective electrode instrument. Analysis of variance showed no statistically significant differences between off-line sample and ISFET Ca++ activities (p greater than 0.05). A second experiment tested the use of the Ca++ ISFET as a control sensor for maintaining a desired Ca++ level. These studies provided a range of Ca++ activity values for comparison of off-line and ISFET values. Infusion rates of sodium citrate or calcium chloride needed to maintain venous Ca++ activity at 1.0 mEq/L or 3.0 mEq/L for periods of 30 min ranged from 440 to less than 1 mg/min during maintenance periods. Observation of hemodynamics during the calcium ion control maneuvers suggests that calcium ion may be used to improve cardiovascular performance during critical care situations that involve cardiovascular instability.

Animals↗

The influence of cervical sympathetic neurons on parathyroid hormone and calcitonin release in the rat: independence of pineal mediation.

The objective of this study was to evaluate the involvement of the pineal gland in modulation of parathyroid hormone (PTH) and calcitonin release found in rats after changes in activity of cervical sympathetic nerves. The response of serum PTH to a hypocalcemia produced by EDTA injection, and of serum calcitonin to a hypercalcemia produced by administering calcium chloride, were studied in rats at the time of the wallerian degeneration of regional sympathetic nerves (i.e., 16 hr after superior cervical ganglionectomy, SCGx). Rats received a pinealectomy or its sham-operation 4 days before SCGx. During wallerian degeneration of nerves after SCGx, a higher hypocalcemia and a lower PTH response were found as compared to sham-SCGx rats, regardless of whether the pineal gland was present or not. When the response of calcitonin to a bolus injection of calcium chloride was assessed, rats subjected to SCGx 16 hr earlier showed a depressed calcitonin release, which was also unaffected after pinealectomy. To a similar extent in pinealectomized and sham-pinealectomized rats, a mild stress given by the subcutaneous injection of turpentine oil brought about a greater hypocalcemia after EDTA, concomitantly with a higher PTH secretory response. In turpentine oil-injected rats, the rise of serum calcitonin was significantly greater than that of vehicle-treated rats, regardless of pineal presence. The results further indicate that cervical autonomic nerves constitute a pathway through which the brain modulates calcium homeostasis and do not support the participation of the pineal gland in short term changes of PTH or calcitonin release.

Animals↗

Studies of mitochondrial calcium movements using chlorotetracycline.

1. The association of calcium with isolated rat liver mitochondrial membranes under various metabolic conditions was monitored using the fluorescent chelate probe, chlorotetracycline. Chlorotetracycline fluorescence increased markedly during energized calcium uptake in the absence of a permeant anion. Uncoupler and a respiratory chain inhibitor caused a rapid decrease in chlorotetracycline fluorescence when added either before or after calcium. During calcium uptake experiments concentrations of calcium exceeding 100 muM caused a transient fluorescence increase followed by an extensive decrease in fluorescence. 2. Changes in the chlorotetracycline-associated fluorescence of the mitochondrial suspensions were correlated with the uptake of exogenous 45Ca. A positive correlation was observed between fluorescence and energized 45Ca uptake in the absence of permeant anions. Addition of the permeant anion, phosphate, caused an extensive decrease in chloretetracycline fluorescence but an enhanced uptake of exogenous 45Ca. 3. The interaction of endogenous mitochondrial calcium with the fluorescent chelate probe was studied under a number of experimental conditions using mitochondria labeled during preparation with 45Ca. Endogenous 45Ca was lost rapidly from the mitochondria upon treatment with uncoupler, antimycin A, and A23187. Potassium phosphate and EGTA had no effect on the endogenous calcium as measured by either the 45Ca content of the mitochondria or the fluorescence of the probe. 4. Mitochondria treated with antimycin A lost most of their endogenous 45Ca within 3 min; subsequent energization of the mitochondria resulted in a partial uptake of the released 45Ca but caused nearly a complete return of the chlorotetracycline fluorescence to the original level. Addition of phosphate did not change the fluorescence level but resulted in an almost complete accumulation of the 45Ca previously released. 5. Following this energized uptake of 45Ca, EGTA, p-trifluoromethoxyphenyl hydrazone of carbonyl cyanide, A23187 and calcium chloride all caused a nearly complete loss of the 45Ca from the mitochondria and, with the exception of calcium chloride, caused an extensive decrease in the fluorescence level. Hence, the apparent location and/or properties of the endogenous calcium in this rat liver mitochondrial system were altered significantly by manipulation of the energetic state of the mitochondrial membrane.

Animals↗

Calcium and calcium-dependent chloride currents generate action potentials in solitary cone photoreceptors.

Vertebrate rod and cone photoreceptors hyperpolarize when illuminated. However, synaptic input from horizontal cells can depolarize cones and even elicit action potentials. Using the whole-cell tight-seal recording technique, we determined that, in solitary cones isolated from a lizard retina, action potentials can be generated by depolarizing current steps under conditions where only two ionic currents are activated. A dihydropyridine-sensitive, inward Ca2+ current that activates at potentials positive to -40 mV can regeneratively depolarize the cell. Subsequently, a SITS-sensitive, Ca2(+)-dependent outward Cl- current repolarizes the cell. We suggest that these ionic currents may help explain lateral inhibition in the retina.

Action Potentials↗

Imaging and diagnostic testing: diastolic dysfunction after coronary artery bypass grafting: a frequent finding of clinical significance not influenced by intravenous calcium.

BACKGROUND: Diastolic dysfunction is common immediately after coronary artery bypass surgery (CABG). The duration of this phenomenon is unknown. Intravenous calcium is frequently administered during separation from cardiopulmonary bypass (CPB). We sought to determine whether intravenous calcium influences perioperative diastolic function and whether diastolic dysfunction persists into the postoperative period. METHODS AND RESULTS: Patients undergoing first-time elective CABG (n = 29) were randomly assigned to receive intravenous calcium chloride (n = 13) or placebo (n = 16) during separation from CPB. Diastolic function was assessed by the pressure-area relation with transesophageal echocardiography and pulmonary capillary wedge pressure (PCWP) measured simultaneously. Left ventricular end-diastolic area (LVEDA) and Doppler indexes were measured at comparable PCWP (within 2 mm Hg) at baseline, after separation from CPB, after sternal closure, and 3 hours after surgery. After CABG, both groups had a significant decrease in LVEDA and mitral E-wave deceleration time that persisted at 3 hours. Because there were no significant differences between the calcium and control groups at any time point, the data for the entire study cohort was analyzed. The LVEDA decreased (stiffness increased) progressively from 16.9 +/- 3.4 cm2 at baseline to 15.8 +/- 2.9 cm2 after CPB, 14.9 +/- 2.5 cm2 after sternal closure, and 14.3 +/- 3.1 cm2 at 3 hours after surgery (P <.0001). The mitral E-wave deceleration time measured at the same time points was 168 +/- 47 ms, 136 +/- 25 ms, 137 +/- 36 ms, and 111 +/- 44 ms (P =.0001). CONCLUSIONS: An increase in left ventricular diastolic chamber stiffness is nearly universal after CABG, and it persists for at least 3 hours after surgery. An intravenous bolus of calcium chloride given during separation from CPB has no measurable negative effect on diastolic function. In the setting of increased chamber stiffness, the PCWP alone does not adequately reflect the volume status and effective preload of the left ventricle.

Aged↗

Inhibition of heart calcium and chloride currents by sodium iodide. Specific attenuation in cAMP-dependent protein kinase-mediated regulation.

The enzymes cAMP-dependent protein kinase (PKA) and protein kinase C (PKC) regulate the activity of cardiac ion channel proteins. In this study the whole-cell arrangement of the patch clamp technique was used to examine the effect of NaI on PKA-stimulated Cl- and Ca2+ channels in isolated guinea pig ventricular myocytes. Cl- currents (ICl) activated either by the beta-adrenergic agonist isoproterenol or the membrane-soluble cAMP analogue, 8-chlorphenylthio (8-CPT) cAMP, were greatly reduced in amplitude after substitution of an external solution containing 140 mM NaCl with a solution containing 140 mM NaI. This reduction was accompanied by a shift of -7 mV in the reversal potential (Erev) for ICl and could be reversed upon return to the NaCl external solution. Inhibition of ICl by NaI occurred in a concentration-dependent manner and was more pronounced for inward ICl (IC50 = 19 mM at -60 mV) than for outward ICl (IC50 = 60 mM at +60 mV). In contrast to ICl activated by PKA, ICl activated by PKC was slightly augmented in the presence of NaI and the Erev was found to shift by -15 mV. Based on these data, the relative permeability of I- to Cl- (PI/PCl) for this channel was calculated to be 1.79. NaI produced no change in the amplitude of inward calcium currents (ICa) recorded under basal conditions, but strongly inhibited ICa augmented by isoproterenol and 8-CPT cAMP, and during dialysis of cells with the catalytic subunit of PKA (CS). The in vitro incorporation of [gamma-32P]ATP into histone IIA and Kemptide, measured in the presence of PKA and cAMP, was not significantly different in assay mixtures containing salts of Cl- and I-. However, the ability of isoproterenol to augment basal ICa in whole-cell experiments was attenuated when experiments were carried out entirely in NaI external solution. Thus, the reduction in ICl and ICa observed in this study may result from a direct effect of I- on the phosphorylation/dephosphorylation of cardiac ion channel proteins or associated regulatory proteins.

Adenosine Triphosphate↗

Assessment of the intrinsic contractile state within an area of myocardium.

Afterload independent, inotropic state sensitive indices of regional function were sought in 31 canine hearts instrumented with piezoelectric crystals to depict trapezoidal areas in the left anterior descending and circumflex arterial beds. Control and postinterventional end-systolic pressure versus regional length and area relationships and regional stroke work versus end-diastolic length and area relationships were inscribed during incremental volume loading on right heart bypass. Hearts were randomized to undergo afterload variation with phenylephrine infusion, contractility augmentation by calcium chloride, or 20 minutes of ischemia in the region of the left anterior descending artery with 30 minutes of reperfusion. All relationships were linear before and after each intervention (mean r = 0.726 to 0.974). The slopes of each correlation were interpreted to quantify intrinsic regional contractility, and all were afterload insensitive (unaffected by phenylephrine). Regional stroke work versus end-diastolic area and length relationships were depressed 55% and 62%, respectively, after ischemia (p less than 0.001 each), whereas neither end-systolic pressure versus regional area nor regional length was significantly altered. Calcium chloride increased regional stroke work versus area 45% in both arterial beds, but significantly increased the other indices only in the left anterior descending and not the left circumflex region. Unlike previous studies of global contractility, correlation of end-systolic events alone did not reliably discriminate perturbations in regional function. The superiority of regional stroke work versus end-diastolic area may be due to incorporating pressure-area changes during the entire cardiac cycle and obviating variability owing to crystal orientation inexactly parallel to fiber shortening.

Animals↗

Correlation between the linearized Frank-Starling relationship and myocardial energetics in the ejecting heart. Afterload independence and sensitivity to inotropic state.

Previous studies suggest that the relationship between end-diastolic volume and stroke work calculated as the area of the pressure-volume work loop is linear, afterload independent, and sensitive to the inotropic state. The correlation of myocardial oxygen consumption with this stroke work could provide an integrated measure of cardiac performance and metabolism to assess perturbations induced by ischemia or pathologic loading conditions. Fourteen canine hearts instrumented for computerized acquisition of instantaneous pressure-volume data and quantitation of myocardial oxygen consumption were studied during progressive volume infusion on right heart bypass (1.5 to 3.5 L/min in 250 ml/min increments). Data acquisition both in the control state and during continuous infusion of calcium chloride (0.03 mEq/kg/min, n = 7) to increase contractility or phenylephrine (2 micrograms/kg/min, n = 7) to alter afterload facilitated the construction of stroke work versus end-diastolic volume and myocardial oxygen consumption versus stroke work relationships by least-squares regression analysis. The cardiac mechanics assessment for this group of dogs confirmed a highly linear (mean r = 0.984) work versus preload relationship that was unaffected by changes in afterload but sensitive to increased contractility (71% increase in slope). The myocardial energetics correlation was also linear (mean r = 0.939) and demonstrated an increased oxygen utilization characteristic of the higher inotropic state produced by calcium chloride infusion (0.047 +/- 0.003 versus 0.070 +/- 0.008 ml oxygen/beat/100 gm left ventricular weight, p = 0.008). Although phenylephrine administration produced variable perturbations of myocardial oxygen consumption, the energetics relationship for this subgroup was not statistically altered by changes in afterload. The features of this cardiac energetics assessment suggest its value as a biological marker to evaluate the postischemic, hypertrophied, or failing heart.

Animals↗

Preparation of collagen modified hyaluronan microparticles as antibiotics carrier.

Hyaluronan (HA), a natural glycoaminoglycan featuring an extracellular matrix, has been suggested as an effective biocompatible material. In this study, the effectiveness of HA microparticles as a carrier system for antibiotics was evaluated, and their physicochemical characteristics were determined. Microparticles were fabricated by the gelation of sulfadiazine (SD) loaded HA solution with calcium chloride through either a granulation (GR-microparticles) or encapsulation (EN-microparticles) process, and atelocollagen was incorporated into the microparticles as an additive in order to improve their physical properties. The characteristics of the microparticles were examined by scanning electron microscopy (SEM), differential scanning calorimetry (DSC), and swelling test. In vitro release experiments were performed for 7 days and the released amount of SD was determined using high-performance liquid chromatography (HPLC). Microscopic observations revealed that the collagen incorporated HA particles had a more compact surface than the HA particles. DSC analysis determined a loss of SD crystallinity in the particles. Calcium chloride retarded the swelling of particles, whereas the loaded drug contents did not affect this property. Both GR-and EN-microparticles sustained SD release with initial bursting effect. SD release from EN-microparticles was faster than from GR-microparticles. In addition, the release rate was dependent on the SD content in the microparticles. These results suggest that collagen modified HA microparticles have a potential as a release rate controlling material for crystalline drugs such as SD.

Anti-Bacterial Agents↗

Mechanical properties and water-vapor permeability of soy-protein films affected by calcium salts and glucono-delta-lactone.

Edible films were prepared from solutions of soy protein with calcium salts and glucono-delta-lactone (GDL). Calcium salts cross-linking interactions with soy-protein isolate (SPI) could result in the formation of films with rigid three-dimensional structure. GDL contributed to the formation of a homogeneous film structure due to increased protein--solvent attraction. Tensile strength (TS) of calcium sulfate treated SPI film (8.6 MPa) was higher than the TS of calcium chloride treated SPI films (6.4 MPa) and the control SPI film (5.5 MPa). Puncture strength (PS) of calcium sulfate treated SPI film (9.8 MPa) was higher than the PS of calcium chloride treated SPI films (8.5 MPa) and the control SPI film (5.9 MPa). SPI film formulated with GDL had larger elongation at break (39.4%) than that of SPI control film (18.2%). Calcium salts and GDL-treated SPI films had lower water-vapor permeability than the SPI control film.

Biofilms↗