Effects of different anticoagulants on determination of erythrocyte glutathione peroxidase.
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Animals↗
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Platelet monoamine oxidase. I: Effect of temperature, anticoagulant, and centrifugation technique.
Platelet yield and monoamine oxidase (MAO) activity per platelet in platelets obtained by the method of Corash, which isolates essentially all platelets from whole blood, and by a single differential centrifugation of whole blood, were compared. Platelets recovered by one spin in a swinging bucket rotor appeared to be a homogeneous population containing a reliable representation of the platelet population obtained by the Corash method since the MAO specific activity yielded by the two methods was not significantly different. The effect of the anticoagulant, temperature, and type of rotor on platelet recovery from blood and MAO activity/platelet were also studied. The MAO and plasma amine oxidaze (PAO) activities obtained from blood anticoagulated with ethylenediaminetetracetate (EDTA) were significantly greater than those found when acid-citrate-dextrose was the anticoagulant. Blood centrifuged in a swinging bucket rotor at room temperature yielded 80-90% of total platelets as compared to the 45-65% recovered with a fixed-angle rotor at 0-4 C. The most efficient method of platelet isolation was found to be a 2 1/2-min room-temperature spin at 600 X g with a swinging bucket rotor using EDTA as anticoagulant.
Adult↗
Histochemical staining. Its use for detection of glyceraldehyde-3-phosphate dehydrogenase in skeletal muscle.
Histochemical staining of glyceraldehyde-3-phosphate dehydrogenase activity in fresh-frozen muscle tissue may be performed using a beta-nicotinamide adenine dinucleotide (beta-NAD) reduced-dehydrogenase tetrazolium-reductase linked reaction. The phosphate-buffered incubating medium contains glyceraldehyde-3-phosphate, beta-NAD, nitro blue tetrazolium, and edetic acid. This technique produces a fine reticular pattern of blue staining in muscle fibers. In human muscle specimens, type I fibers stain darker than type II fibers.
Animals↗
Carboplatin in combination with paclitaxel in advanced ovarian cancer: dose determination and pharmacokinetic and pharmacodynamic interactions.
Data from various phase I/II studies of carboplatin in combination with paclitaxel (Taxol; Bristol-Myers Squibb Company, Princeton, NJ) have suggested that the degree of thrombocytopenia seen is less than that expected when carboplatin is given alone. However, some studies also have suggested that the area under the plasma concentration-time curve (AUC) of carboplatin is lower than that expected, raising the possibility of a pharmacokinetic interaction. Patients with advanced epithelial ovarian cancer were treated with first-line carboplatin (AUC = 7, using the 51Cr EDTA [edetic acid] clearance method) and escalating doses of paclitaxel. Thrombocytopenia was mild and was significantly less when the paclitaxel dose was 175 mg/m2 versus 150 mg/m2. Paclitaxel kinetics were nonlinear, as previously reported. The achieved carboplatin AUC was 7 +/- 1 mg/mL.min, indicating that the pharmacokinetics of carboplatin are not affected by paclitaxel. Glomerular filtration rates measured in 184 patients using the 51Cr EDTA clearance method were compared with rates estimated from the plasma creatinine level using the Cockcroft-Gault or Jeliffe formulas and showed a significant bias of these two formulas. Clearances above 50 mL/min were underestimated by an amount that became greater as the clearance increased and was approximately 25% to 35% for patients with clearances in the normal range. Since creatinine-based methods have been used in many previous studies, care is needed in interpreting the predicted AUC values from these studies. Carboplatin and paclitaxel may be given safely in combination at full doses, and the thrombocytopenia seen is significantly less than that observed with single-agent carboplatin. No evidence exists of a pharmacokinetic interaction, and the observation in some studies that the carboplatin AUC was lower than expected was probably due to the methodology used to estimate the glomerular filtration rate.
Adult↗
Green hair.
Green hair is an unusual dermatologic condition usually due to the deposition of copper from exogenous sources. We report the cases of two patients who presented to our clinic with green discoloration of their hair. This pigmentation of hair has generally been reported in patients with blond hair as a consequence of increased concentrations of copper in domestic or swimming pool water. Although an increased copper content of the affected hair seems to be a prerequisite, other predisposing factors have to be present for this situation to occur. These include previous hair damage (mechanical, sun exposure, bleaching, dyeing, waving), frequent contact with chlorinated water, or use of alkaline shampoos. Several options for treatment have been described for this problem, including application of hot vegetable oil, hydrogen peroxide, edetic acid- or D-penicillamine-containing shampoos, or hydroxyethyl diphosphonic acid.
Adult↗
Periodontal tissue-vitality after different etching modalities.
The purpose of the present study was to investigate possible immediate necrotizing effects on periodontal tissues of etching agents operating either at low or neutral pH. Phosphoric and citric acids, both of which operate at low pH, exerted immediate (within 20 s) necrotizing effects on both mucosal flaps and periodontal tissues. The penetration depth increased with time up to 1/4 of the circumference of the root after 3 min. This was in sharp contrast to EDTA which operates at neutral pH. EDTA failed to induce any detectable necrosis during the experimental periods. Whether or not the immediate necrotizing effects of low pH etching agents also impair periodontal healing in the long-term perspective, needs to be evaluated with controlled experimental studies in vivo as well as clinically in order to determine whether etching at neutral pH is preferable.
Acid Etching, Dental↗
[Effect of sodium suberogorgin on hemodynamics and myocardial ischemic-reperfused injury in isolated guinea pig working hearts].
AIM: To study the actions of sodium suberogorgin (Sub) on normal- and myocardial ischemic-reperfused-working hearts. METHODS: The isolated guinea pig working hearts were perfused with edetic acid (0.2 mumol.L(-1))-Krebs-Henseleit solution by left pulmonary vein at 37 +/- 0.5 degrees C. The perfusing pressure of left atrium and the afterload of left ventricle were maintained at 1.0 and 7.2 kPa, respectively. During 50-min low-flow global ischemia, coronary flow was maintained at 4.7 +/- 0.2% of normal working hearts via retrograding perfusion. Reperfusion lasted 35 min following ischemia. RESULTS: Before ischemia, the aortic pressure, left ventricular systolic pressure, the maximal rate of left ventricular pressure change (+ dp/dtmax and -dp/dtmax), cardiac output, stroke volume, left ventricular end-diastolic pressure, and heart rate of working hearts treated with Sub 10 mumol.L(-1) for 10 min were increased by 14%, 17%, 17%, 22%, 15%, 32%, -200%, and -11%, respectively. These parameters were reduced when Sub was increased to 50 mumol.L(-1). Reperfusion aggravated the myocardial damages induced by ischemia. Sub 10 mumol.L(-1) used before and during low-flow ischemia completely restored all above parameters except heart rate to the level of preischemia. When Sub was increased to 50 mumol.L(-1), only an attenuation of myocardial damage was observed. CONCLUSION: Sub obviously improved the ischemic-reperfused injury in isolated working hearts.
Animals↗
CD13 (GP150; aminopeptidase-N): predominant functional activity in blood is localized to plasma and is not cell-surface associated.
This study is the first to report the presence of CD13/glycoprotein 150 (GP150)/aminopeptidase-N activity in cell-free plasma. We have determined that aminopeptidase-N in plasma provides, quantitatively, aminopeptidase-N's predominant functional activity within flowing blood. Thus, while aminopeptidase-N activity observed in whole blood can be partly, but significantly, blocked by the CD13 monoclonal antibody (MAB) WM15, the magnitude of such inhibition is low (< 25%) and similar to that observed using washed cell fractions selectively enriched for neutrophils (30.6% inhibition) or monocytes (21.8% inhibition). Plasma, free of cell components, possesses substantial aminopeptidase-N activity that is largely inhibitable (> 70%) by WM15. Blood collected into heparin or citrate yields similar data, while blood collected into EDTA gives rise to reduced CD13/aminopeptidase-N activity, consistent with inhibition of the known heavy-metal ion association necessary for proper functioning of this molecule. Although monocyte- and granulocyte-enriched cell fractions possess aminopeptidase-N activity significantly inhibitable by CD13 antibodies, lymphocyte-enriched cell fractions also possess aminopeptidase-N-like activity; however, in the latter case, this activity is not inhibitable by CD13 antibodies. Immunoaffinity isolation of plasma aminopeptidase-N has also been carried out; further characterization using functional studies and sodium dodecyl sulfate-polyacrylamide gel (SDS-PAGE) electrophoresis indicates that CD13 MABs can completely clear plasma of aminopeptidase-N activity and that the purified protein has similar electrophoretic characteristics to cell-derived material. These data, therefore, provide evidence for the presence within blood both of a soluble (that is, non-cell-associated) form of CD13/GP150/aminopeptidase-N localizable to plasma and of cell-associated, aminopeptidase-N-like proteins other than CD13/GP150. These findings have significant implications for our understanding of the many functions of this molecule in blood.
Aminopeptidases↗
Isolation of the soluble substrate recognition component of the dicarboxylate transport system of Escherichia coli.
A soluble, periplasmic protein was isolated from Escherichia coli cells by chromatography on columns of aspartate-coupled Sepharose 4B. This protein has a molecular weight of about 15,000 and, as judged by competition experiments, binds the three dicarboxylic acids, succinate, malate, and fumarate and, addition, the monocarboxylic acid D-lactate. The periplasmic protein seems to be missing from some mutants of E. coli (designated cbt) which are incapable of transporting succinate in whole cells.
Bacterial Proteins↗
Blood lead - an inadequate measure of occupational exposure.
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Adult↗
The routine measurement of platelet size using sodium citrate alone as the anticoagulant.
Mean platelet volume (MPV), a measure of platelet size, is becoming recognised as an important marker of platelet function. However, platelets swell in edetic acid (EDTA), the standard haematology anticoagulant, in a time-dependent manner making such measurements potentially unreliable. The effect of incubation time on MPV, and platelet distribution width (PDW), as measured in EDTA, low (1:9 volume/volume with blood) or high (1:4 v/v with blood) concentration sodium citrate was studied. MPV measured in high concentration sodium citrate did not change with time in contrast to MPV measured in either low concentration sodium citrate or EDTA which both increased in an inverse exponential fashion. MPV and PDW, measured in high concentration sodium citrate, had similar within-assay and between-assay coefficients of variation as other platelet, red cell and white cell haematology variables measured in EDTA: MPV 1.4%, 2.1%; PDW 1.4%, 1.5%; MCV 0.4%, 0.7%; PC 3.1%, 6.1%; WCC 1.5%, 7.3%; Hb 2.1%, 2.4% respectively. MPV measured in EDTA and corrected for incubation time approximated to, but was higher than, the MPV measured in high concentration sodium citrate. PDW correlated inversely with platelet count (r = -0.415, 2p < 0.001). MPV may be measured in sodium citrate (at 1:4 v/v with blood) alone with a better accuracy and reproducibility than similar measurements made in EDTA. Furthermore, such measurements are not influenced by incubation time, unlike for EDTA.
Anticoagulants↗
[Controlled therapy of lead poisoning].
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Acute Disease↗
Effect of porphyrin and irradiation on heme biosynthetic pathway in endothelial cells.
To study porphyrin biosynthesis in human umbilical vein endothelial cells (EC), cultured EC were incubated with delta-aminolevulinic acid (ALA), the precursor of porphyrin synthesis, and porphyrins generated were measured spectrofluorometrically. EC accumulated porphyrins in an incubation time-dependent and ALA dose-dependent fashion. Porphyrin accumulation was enhanced by CaMg EDTA, a ferrochelatase inhibitor, suggesting the participation of ferrochelatase. The effect of porphyrin and irradiation on porphyrin accumulation was evaluated by exposing porphyrin-containing EC to the UVA and blue light, followed by incubation with ALA for 2-48 h. Decreased porphyrin accumulation 2-48 h after the irradiation was noted at all the time-points examined. However, no alteration of ferrochelatase activity was observed, suggesting that the decrease in porphyrin levels was due to suppression of the activity of the other enzymes in the earlier steps of the heme biosynthetic pathway.
Aminolevulinic Acid↗
Periodontal regeneration: root surface demineralization.
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Animals↗
[Long term biokinetics of 169Yb following injection as citrate, hydroxyethylethylenediaminetetraacetate and nitrilotriacetate].
169Yb complexes with known biokinetics in tumour-bearing mice up to 48 h p.i. were injected into healthy mice to study the radionuclide biodistribution in various organs and tissues for 672 h after injection, in order to obtain reliable biokinetic data in an animal model, not affected by tumour-growth, as a basis for the calculation of biological half-life and dose distribution. The results demonstrated the existence of at least two components with different biological half-lives in the organs and tissues investigated. The effective half-lives of these components decreased with increasing stability of the complexes administered. The effective half-life of the fast component was a few hours and that of the slow one between about 200 and 800 h.
Animals↗
[Determination of the survival of Trypanosoma evansi in equine blood, using the microhematocrit method].
The microhaematocrit (MH) technique was used to study the survival of Trypanosoma evansi in blood from two herds of naturally-infected horses. A comparison was made between samples treated with ethylenediaminetetraacetic acid and sodium citrate (alone or with 1% glucose), and sent to the laboratory packed in ice. In general, the number of samples yielding positive results by the MH technique showed the least variation during the first 24-36 h after sample collection. Survival varied with the anticoagulant used, but it declined rapidly from 48 h after collection, although live parasites were still observed in up to 10% of samples until the seventh day. On the basis of the results obtained, the authors recommend the use of sodium citrate in treating equine blood samples for the parasitological diagnosis of T. evansi.
Animals↗
Measurement of lipoprotein (a): comparison of Macra and Imubind methods.
Our specific aim was to compare lipoprotein(a) [Lp(a)] measured by two enzyme-linked immunosorbent assays (the Strategic Diagnostics, Inc. Macra Lp(a) and the American Diagnostica, Inc. Imubind Lp(a) Stripwell) in sequentially referred hyperlipidemic patients. The Macra method binds Lp(a) in serum or plasma to monoclonal anti-Lp(a) in microtiter test wells followed by polyclonal anti-Lp(a) and a chromogenic reaction; the Imubind method is similar but uses two polyclonal anti-Lp(a) antibodies. Within run coefficients of variation were 1.5 percent for the Macra (Lp[a] 27.2 +/- 0.4 mg/dl) and 3.4 percent for the Imubind method (Lp[a] = 18.9 +/- 0.6 mg/dl). Between-run coefficients of variation for the Macra method were 4.7 percent (Lp[a] 14.8 +/- 0.7 mg/dl) and 8.3 percent (Lp[a] of 33.5 +/- 2.8 mg/dl). For the Imubind method in nine separate analytical runs, the between run coefficients of variation were 8.4 percent (Lp[a] of 15.4 +/- 1.3 mg/dl), 3.0 percent (18.8 +/- 0.6 mg/dl), and 5.7 percent (25.6 +/- 1.5 mg/dl). The intraclass correlation was 0.92 (p < or = 0.0001) for duplicate aliquots (n = 210) quantitated by both methods, and the lower limit of the 95 percent confidence interval of the intraclass correlation was 0.90. Comparison of the two methods revealed no systematic bias (p = 0.09), since the lower limit of the 95 percent intraclass correlation confidence interval was > or = 0.75, the two methods for measuring Lp(a) are considered interchangeable. Given the importance of Lp(a) as an independent risk factor for coronary heart disease, it is clinically important to have precise and accurate methods for its measurement.
Adolescent↗
Peroxide induced chemiluminescence in an in vitro proline hydroxylation system.
This communication describes a hydrogen peroxide (HOOH) induced chemiluminescence (CL) in an in vitro aromatic (proline) hydroxylation system. The reactive components of the system are ascorbic (AA), ethylene diamine tetraacetic disodium salt (EDTA), ferrous sulfate, and HOOH. The CL is (1) nearly dissipated within three minutes, (2) enhanced and/or sustained by proline and polylysine to a greater degree than by alanine, (3) partially inhibited by a,a' dipyridyl, EDTA, and ethanol, (4) most dependent upon the presence of Fe2+, AA, and HOOH. The in vitro proline hydroxylation system is more effective than ground state oxygen in terms of the CL produced and the percent of hydroxyproline formed.
2,2'-Dipyridyl↗