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PubMed1983

An experimental model for measuring intestinal permeability.

In the nonanesthetized rat, the jejunal permeability to four simultaneously administered molecules, mannitol, phenol red, inulin and PVP, was measured by analyzing blood, serum, urine and duodenal fluid for these compounds. Of the molecules which had entered the body, approximately 50% were found in the urine, another 50% in the extracellular space and only about 1% were excreted into the duodenal juice. The intracellular content of the molecules is not accounted for in these numbers. The rate of permeation decreased with increasing molecular weight of the substances. EDTA (25 mmol/l) and deoxycholate (5 mmol/l) increased the jejunal permeability for these molecules but not for mannitol. The alterations in mucosal cell turnover and morphology induced by hydroxyurea did not change jejunal permeability for mannitol and phenol red at any time. 24 and 48 h following hydroxyurea, jejunal permeability for inulin and PVP was decreased.

Animals↗
PubMed1975

Immunological comparison of various human pregnancy-associated plasma proteins.

Direct immunodiffusion comparison with specific antisera demonstrated that all of the four pregnancy-associated plasma proteins (PAPPs) described in our laboratory are distinct from the pregnancy zone protein (von Schoultz); alpha2-pregnoglobulin (Berne); pregnancy-associated alpha2-glycoprotein (SP3) (Bohn); new serum alpha2-macroglobulin (Stimson); PAG (Horne); pregnancy-associated alpha2-globulin (Kasukawa); Pal (McLaren), and Xh protein (Dunston). All the latter proved to be immunologically idnetical to each other, and apparently represent the same protein described under different names. It was confirmed that none of the PAPPs is immunochemically related to placental alkaline phosphatase. PAPP-A, PAPP-C, and the pregnancy zone protein, but not HPL (PAPP-D), showed a decreased anodic mobility when treated with neuraminidase; their reactivities with antibody were essentially unaffected by the ezyme, however. Certain detergents had no effect on the immunological reactivities of the PAPPs and the pregnancy zone protein in whole plasma, while butanol and desoxycholate partially, and urea, SDS and cetylpyridinium largely inactivated them.

Alkaline Phosphatase↗
PubMed1991

Inhibition of acid production by Streptococcus mutans NCTC 10449 by zinc and the effect of metal speciation.

Ionic zinc as zinc sulphate was strongly inhibitory to acid production from glucose by washed cell suspensions of Streptococcus mutans NCTC 10449 in a pH-stat assay (50% inhibition at 0.1 mM zinc). Zinc was adsorbed to the cells (up to 8 micrograms/mg cells). Several other zinc salts that dissociated to free zinc ions were also strongly inhibitory. Zinc in a partially complexed form as zinc citrate was less strongly inhibitory (50% inhibition at 0.2 mM zinc), and adsorption of zinc was lower (up to 3 micrograms/mg cells). Stoichiometric addition of ethylenediaminetetra-acetic acid (EDTA) to assays containing zinc sulphate completely removed inhibition and adsorption of zinc. A good correlation between inhibition of acid production and adsorption of zinc was found, and zinc adsorption was correlated with the levels of free zinc ion in the assay mixtures. The strongly anionic complexes of zinc, Zn(CIT)-, Zn(CIT)4-(2), and Zn(EDTA)2-, were not inhibitory and did not adsorb to the bacterial cells.

Acids↗
PubMed1978

Critical study of the value of renal clearances measured by the single shot technic.

A critical approach to the problem of establishing new methods of measurement of kidney function is presented. Variability of the reference clearances inulin and PAH is large, thus reference method ratios close to 1 are not sufficient to ascertain the value of the method under investigation. EDTA 51Cr and hippuran 125I, single shot technics illustrate the need for more accurate criteria required for validating a new technic.

Aminohippuric Acids↗
PubMed1988

Combined treatment of medullary sponge kidney by EDTA potassium citrate and extracorporeal shock wave lithotripsy.

A case of successful renal calculus dissolution by the combined treatment which consists of irrigation with ethylenediaminetetraacetic acid (EDTA), potassium citrate, and extracorporeal shock-wave lithotripsy (ESWL) is described here. Renal irrigation via nephrostomy, which was the main treatment, was attempted on a 34-year-old Japanese male who had bilateral nephrocalcinosis caused by type 1 renal tubular acidosis associated with an impacted calculus in the right ureter. Finally, most of the calculi have been dissolved within 1 year.

Adult↗
PubMed1975

Hemoglobin Syracuse (alpha2beta2-143(H21)His leads to Pro), a new high-affinity variant detected by special electrophoretic methods. Observations on the auto-oxidation of normal and variant hemoglobins.

Family members from four generations were found to have polycythemia and increased whole blood O2 affinity (P50; 11 mm Hg; normal, 27 mm Hg). No abnormal hemoglobin bands were seen after electrophoresis on starch gel at pH 8.6 or agar gel at pH 6.0. Analysis of the oxygenated hemolysate by isoelectric focusing on polyacrylamide gel revealed two closely spaced bands. When deoxygenated hemolysate was analyzed in oxygen-free gels, the two components were more widely separated. About 40% of the patient's hemoglobin focused at a more acid pH than hemoglobin A, indicating a hemoglobin variant with impaired Bohr effect. Chromatography of globin in 8 M urea revealed two beta-chain peaks, the first of which was eluted at a lower buffer molarity than normal beta chain. Fingerprints of tryptic digests of the aminoethylated chains were done on silica gel thin-layer plates. Tp 14 from the abnormal beta chain had slower electrophoretic mobility and a greater Rf value. Amino acid analyses of this peptide gave values identical with those of betaTp 14, except that it contained one proline residue and no histidine. Since the one His in betaTp 14 is in position 143, hemoglobin Syracuse in alpha2beta2-143 His leads to Pro. Native Hb Syracuse could be separated from hemoglobin A on a carboxymethylcellulose column. The inclusion of 0.1 mM EDTA in the preparative buffers proved very useful in reducing the formation of methemoglobin. Oxygen equilibria of purified hemoglobin Syracuse showed high oxygen affinity (P50 value 12% that of hemoglobin A) and lack of cooperativity between subunits (Hill's n equals 1.1). The alkaline Bohr effect was about half that of hemoglobin A. The proline substitution at betaH21 disrupts the helical configuration and probably prevents the formation of salt bonds that are important in stabilizing the deoxy structure and contribute to the alkaline Bohr effect. Since beta143 His is a binding site for 2,3-diphosphoglycerate (2,3-DPG), it is not suprising that hemoglobin Syracuse had markedly impaired reactivity with 2,3-DPG. Hemoglobin Syracuse auto-oxidized more slowly than hemoglobin A, probably reflecting a slower rate of dissociation of oxygen from fully liganded hemoglobin.

Adult↗
PubMed1976

The purine nucleotide cycle. A pathway for ammonia production in the rat kidney.

Particle-free extracts prepared from kidney cortex of rat catalyze the formation of ammonia via the purine nucleotide cycle. The cycle generates ammonia and fumarate from aspartate, using catalytic amounts of inosine monophosphate, adenylosuccinate, and adenosine monophosphate. The specific activities of the enzymes of the cycle are 1.27+/-0.27 nmol/mg protein per min (SE) for adenoylosuccinate synthetase, 1.38+/-0.16 for adenylosuccinase, and 44.0+/-3.3 for AMP deaminase. Compared with controls, extracts prepared from kidneys of rats fed ammonium chloride for 2 days show a 60% increase in adenylosuccinate synthetase and a threefold increase in adenylosuccinase activity, and a greater and more rapid synthesis of ammonia and adenine nucleotide from aspartate and inosine monophosphate. Extracts prepared from kidneys of rats fed a potassium-deficient diet show a twofold increase in adenylosuccinate synthetase and a threefold increase in adenylosuccinase activity. In such extracts the rate of synthesis of ammonia and adenine nucleotide from aspartate and inosine monophosphate is also increased. These results show that the reactions of the purine nucleotide cycle are present and can operate in extracts of kidney cortex. The operational capacity of the cycle is accelerated by ammonium chloride feeding and potassium depletion, conditions known to increase renal ammonia excretion. Extracts of kidney cortex convert inosine monophosphate to uric acid. This is prevented by addition of allopurinol of 1-pyrophosphoryl ribose 5-phosphate to the reaction mixture.

AMP Deaminase↗
PubMed1994

Comparison of absorption-enhancing effect between sodium caprate and disodium ethylenediaminetetraacetate in Caco-2 cells.

The effects of sodium caprate (C10) and disodium ethylenediaminetetraacetate (EDTA) on the epithelial permeability of polyethylene glycol 4000 (PEG4000) were examined in Caco-2 cell monolayers on the polycarbonate membrane (filter) of a Transwell cluster dish. Under serosal Ca(2+)-free conditions irrespective of the presence or absence of mucosal Ca2+, the permeability of PEG4000 was remarkably enhanced, even in the absence of enhancer. For control experimental conditions with a permeation enhancer in Caco-2 cells, Ca2+ was added to both the mucosal and serosal sides. Under the control conditions, EDTA increased the permeability of PEG4000 to 14 times the control and C10 by 3.5 times. Mucosal Ca(2+)-free conditions further increased the EDTA-enhancing effect, to 29 times the control value. In contrast to the EDTA effect, however, the C10 effect was unchanged in the absence of mucosal Ca2+. These results suggest that the EDTA effect is induced by chelation with Ca2+ but that C10 has some other action in addition to chelation.

Calcium↗
PubMed2005

The effect of clodronate on the integrity and viability of rat small intestine in vitro-a comparison with EDTA.

Although substances, which can increase the paracellular permeability of intestinal mucosa, could be very helpful for increasing the bioavailability of hydrophilic drugs, they are not used therapeutically due to the possibilities of acute or long-term toxicity (intestinal inflammations due to penetration of bacterial fragments into subepithelial spaces). In this paper the abilities of a calcium chelator EDTA and clodronate (a first generation bisphosphonate) to increase the paracellular permeability were assessed using rat jejunum in side-by-side diffusion chambers while the viability of the tissue was monitored by transepithelial potential difference. Although clodronate is less potent than EDTA in depleting calcium from the intestinal tissue, it significantly increased the paracellular permeability of viable rat jejunum "in vitro" when tested at 15 mM and higher concentrations (the highest therapeutic dose dissolved in 250 ml gives a 22 mM solution of clodronate). This effect was reversible under "high-calcium" conditions. Since clodronate therapy does not have any long-term consequences it was concluded that a safe, transient increase of small intestinal permeability is possible. However, the acute gastrointestinal undesired effects, which can develop during the therapy with high doses of clodronate, might also occur after oral applications of paracellular permeability enhancers. Namely, 30 mM and higher concentrations of clodronate caused a loss of the tissue viability in all rat jejunal segments tested in "in vitro" conditions. A similar effect was observed with much lower concentrations of EDTA.

Animals↗
PubMed1983

Renin release and lipid peroxidation by ascorbic acid in the renin granule fraction of rat kidney cortex.

This study was carried out to investigate the effect of lipid peroxidation in the renin granule fraction on renin release from the granules. Ascorbic acid was used to cause lipid peroxidation in the renin granule fraction prepared from rat kidney cortex homogenate. Renin activity was measured by radioimmunoassay and lipid peroxidation was estimated by means of the thiobarbituric acid test. Ascorbic acid, at the concentrations from 5 to 100 microM, produced a dose-dependent increase in lipid peroxidation during incubation of the renin granule fraction at 37 degrees C for 30 min, accompanied by increased release of renin from the granules. On the other hand, dehydroascorbic acid showed no effects on lipid peroxidation and renin release. The simultaneous increases in lipid peroxidation and renin release induced by ascorbic acid in the renin granule fraction were markedly suppressed by the addition of disodium ethylenediaminetetra-acetic acid and antioxidants such as N,N'-diphenyl-p-phenylenediamine and hydroquinone. These findings indicate that lipid peroxidation in the renin granule fraction results in the stimulation of renin release from the granules.

Animals↗
PubMed2002

Determination of the length of sedimentation reaction (erythrocyte sedimentation rate) in non-anticoagulated blood with the Microtest 1.

The erythrocyte sedimentation rate (ESR), now more appropriately referred to as the "length of sedimentation reaction in blood (LSRB)", remains the most widely used laboratory test for monitoring the course of infections, inflammatory diseases and some types of cancer. Thanks to the several recently developed methods for the measurement of this reaction, the safety and reliability of LSRB testing procedures have improved. The method for LSRB measurement recommended by the International Council for Standardization in Hematology (ICSH) and the National Committee for Clinical Laboratory Standards (NCCLS) is based on the traditional Westergren method, using EDTA-anticoagulated samples. The present paper describes and evaluates a procedure for LSRB measurement with a new manual system, the Microtest 1, which requires only 30 microl of blood and is optimal for pediatric use. Microtest 1 results correlated satisfactorily with the ICSH recommended method (r=0.88, 95% CI 0.84-0.91; y=3.45+0.85x), had no significant bias (2.15, 95% CI -0.32-4.63) and had an imprecision of less than 7% for the reference range values. In addition, the results obtained with the Microtest 1 in "native" whole blood were comparable with those obtained with the reference method in K3EDTA and sodium citrate-anticoagulated specimens; (bias = 1.19, 95% CI -1.78 to 4.16 for K3EDTA-anticoagulated samples and bias = 2.43, 95% CI -0.58 to 5.44 for sodium citrate-anticoagulated samples).

Adult↗
PubMed2006

Effects of preanalytical variables on peptide and protein measurements in human serum and plasma: implications for clinical proteomics.

There is a wealth of knowledge in the field of in vitro diagnostics with regard to preanalytical variables and their impact on the determination of peptide and protein analytes in human serum and plasma. This information is applicable to clinical proteomics investigations, which utilize the same sample types. Studies have demonstrated that the majority of variations and errors in in vitro diagnostics seem to occur in the preanalytical phase prior to specimen analysis. Preanalytical processes include study design, compliance of the subjects investigated, compliance of the technical staff in adherence to protocols, choice of specimens utilized and sample collection and processing. These variables can have a dramatic impact on the determination of analytes and can affect result outcomes, reproducibility and the validity of investigations. By drawing analogies to in vitro diagnostics practices, specific variables that are likely to impact the results of proteomics studies can be identified. Recognition of such variables is the first step towards their understanding and, eventually, controlling their impact. In this article, we will review preanalytical variables, provide examples for their effects on the determination of distinct peptides and proteins and discuss potential implications for clinical proteomics investigations.

Anticoagulants↗
PubMed1983

A scanning electron microscope study of the effects of various agents on instrumented periodontally involved root surfaces.

This study evaluated the effects of chemical and biologic agents on periodontally diseased root surfaces which had been scaled or root planed. The proximal surfaces of 25 teeth were scaled to remove all visible calculus, and the proximal surfaces of another 25 teeth were vigorously root planed to remove all cementum and to achieve a hard, smooth, glass-like surface. Five scaled and five root planed specimens were randomly selected for light microscopic examination to determine the amount of cementum removed. Cementum remained on all scaled surfaces but root planing had removed most of the more coronal cementum. Each of the 40 remaining teeth (20 scaled and 20 root planed) was longitudinally sectioned to obtain an experimental and control specimen. Four scaled and four root planed sections were randomly selected as experimental specimens for a test of each of the following five agents: (1) saturated citric acid for three minutes; (2) 15% EDTA for 5 minutes; (3) sodium hypochlorite for 5 minutes, followed by a 30-second application of 5% citric acid; (4) sodium hypochlorite alone for 5 minutes; and (5) 2% sodium deoxycholate (NAD) for 1 minute, followed by a 1-minute rinse in distilled water, and then a 1-minute application of 5% Cohn's fraction IV1. The control for each experimental specimen was treated with saline. All samples were prepared for SEM and examined at 3,000 X. Areas of particular interest were also examined at 12,000 X. The chemical treatments exposed only individual collagen fibers or irregular fiber bundles on the scaled surfaces. Saturated citric acid, EDTA, and sodium hypochlorite with citric acid neutralization removed debris and exposed openings in the root surfaces. Sodium hypochlorite alone and NaD/Cohn's fraction IV1 were less effective in removing surface debris and had an effect similar to that seen in the saline controls. Application to root planed specimens of saturated citric acid, EDTA, and sodium hypochlorite followed by 30 seconds of citric acid neutralization resulted in surfaces virtually free of debris and with numerous collagen fibers exposed on the surface. EDTA appeared to cause a morphologic change in the collagen fibers. Sodium hypochlorite alone, sodium deoxycholate followed by Cohn's fraction IV1, and physiologic saline were relatively ineffective in surface debridement.

Adult↗
PubMedDate not supplied

Light-catalyzed chromium(VI) reduction by organic compounds and soil minerals.

Detoxification of Cr(VI) through reduction has been considered an effective method for reclaiming Cr-contaminated soil, sediment, and waste water. Organic matter is widely distributed in soil and aquatic systems; however, low Cr(VI) reduction rates inhibit the adoption of Cr reduction technologies by industry. Scientists have been aware of Cr(VI) reduction catalyzed by soil minerals; however, most of the studies focused on using semiconductors as catalysts with UV irradiation to accelerate the redox reactions. The objective of this study was to evaluate the rates of Cr(VI) reduction by fluorescence light in the presence of organic materials with or without specific soil minerals. Experimental results showed that dissolved organic compounds reduced Cr(VI) slowly under laboratory light; however, Cr(VI) reduction was greatly enhanced when growth chamber light was applied. Low photon flux (i.e., laboratory light) only enhanced Cr(VI) reduction by organics when Fe(III) was also present, because the Fe(II)-Fe(III) redox couple accelerated electron transfer and decreased electrostatic repulsion between reactants. Laboratory light was required to initiate Cr(VI) reduction catalyzed by TiO2; nonetheless, light-catalyzed Cr(VI) reduction by smectite and ferrihydrite could occur only when greater light energy was provided with a growth chamber light. Our results suggest a potential pathway for Cr(VI) reduction using naturally occurring organic compounds and colloids in acidic water systems or in surface soils when light is available.

Catalysis↗
PubMed2000

Iomeprol: a review of its use as a contrast medium.

UNLABELLED: Iomeprol is a nonionic, monomeric iodinated contrast medium. Unlike the older ionic agents, iomeprol has low chemotoxicity, osmolality and viscosity and high water solubility. Compared with other nonionic contrast media, the osmolality and viscosity are lower and the water solubility is reported to be higher with iomeprol. Most radiographs (about 67 to 100%) obtained with iomeprol (containing 150 to 400 mg/ml of iodine) were of good or excellent quality in noncomparative and comparative trials recruiting 40 to 6127 patients undergoing various radiographic procedures. As expected, the diagnostic efficacy of iomeprol did not differ significantly from that of other nonionic agents (iopamidol, iopromide, iohexol and iotrolan). Iomeprol (containing 150 to 400 mg/ml of iodine) was well tolerated in clinical trials. Most adverse events were transient and of mild to moderate intensity and were similar to those observed with other contrast media. The overall incidence of adverse events ranged from 3 to 49.7% and mainly included localised pain (< or =6%) and heat sensations (8 to 45%), taste disturbances (3 to 27%) and various pseudoallergic reactions (< or =20% for each type of event). The incidence of heat or pain and taste disturbances with iomeprol was similar to that observed with iopromide and iopamidol. Pain (but not heat sensations) was reported significantly less frequently and taste disturbances reported significantly more frequently with iomeprol than with iohexol in a comparative trial. Pseudoallergic reactions (such as nausea, vomiting, skin reactions, dizziness, headache) were significantly less common with iomeprol than with ioxaglate and occurred at a similar frequency to that with iopromide and iopamidol. Cardiovascular events were rarely observed with iomeprol. Currently available iomeprol solutions contain a range of iodine concentrations (150 to 400 mg/ml) and are approved for a wide variety of diagnostic procedures. Iomeprol solutions are chemically stable which negates the need for chelating agents. Formulations of this agent are therefore the first not to contain edetic acid (EDTA). CONCLUSIONS: Iomeprol shows equivalent diagnostic efficacy, and a similar adverse event profile, to that of other nonionic contrast media. The availability of a range of iodine concentrations enables iomeprol to be used in a variety of diagnostic procedures. Iomeprol, like others in its class, is suitable for use in diagnostic imaging.

Contrast Media↗