Isotope effect in the binding of tritium and 14C-labelled cortisol to corticosteroid-binding-globulin in serum

We have determined the free cortisol concentration in serum using either the Amicon MPS-1 ultrafiltration-centrifugation method (I) or equilibrium dialysis (II). If procedure I was used we found that [1,2,6,7-3H]-, and [4-14C]cortisol had a lower affinity than unlabelled cortisol for corticosteroid binding globulin (CBG). The binding affinity (Ka) to three separate CBG-containing samples was 8-18 times lower for [1,2,6,7-3H]cortisol and 30-90 times lower for [4-14C]cortisol, when compared with that of unlabelled cortisol. This difference in affinity to CBG was not observed if method II was used for the free cortisol determinations. The observed isotope effect in method I is not caused by unspecific binding to material such as the Amicon MPS-1 chamber or to impurities in the tracer. We suggest that the centrifugation step during ultrafiltration changed the conformation of CBG, thereby reducing its affinity for labelled cortisol. It is concluded that incorrect results will be obtained if radiolabelled is cortisol used for determining the free cortisol content of plasma with the Amicon MPS-1 device.     

Analysis of electrical phenomena accompanying the growth of Neurospora crassa hyphae: theory and experiment

To describe electrical phenomena observed in growth of Neurospora crassa hyphae, a theoretical model was developed considering the hypha as a one-dimensional electric cable with non-uniform longitudinal distribution of current sources reflecting the activity of proton pumps. A profile of the density of the pump current along the hypha is proposed, at which the results of simulation quantitatively coincide with the results of physiological experiments. The model values of energy coupling in the growth zones were estimated. The experimental dependence of the elongation rate of regenerating apical hypha fragments on their lengths was determined. Based on the comparison of these experimental results with the results of analysis of the model, the contribution of the axial metabolite transport, from the distal parts of the hypha to the apical part, to the dynamics of the apical cell growth was estimated. The possibility of evaluating the intensity of high-molecular-weight syntheses and/or accumulation of substances in granules was demonstrated. The growth rate of the regenerating hypha fragments was shown to correlate with the electric current flowing into the apical fragment 0.2-mm in length.     

Uptake and metabolism of lipoprotein-X in mesangial cells

An HPLC method developed to detect in a single run both atenolol and chlorthalidone, extracted from plasma, using two detectors (UV for chlorthalidone and fluorometric for atenolol) connected in series, is described. The drugs were separated on an ODS column at room temperature using a 0.05 M sodium dodecyl sulphate in phosphate buffer (pH 5.8)-n-propanol (95:5, v/v) solution, delivered at a flow-rate of 1.3 ml/min. Having ascertained the sensitivity (10 ng/ml of both drugs) and the intra-day reproducibility (pre-study validation), the reliability of the method was verified by inter-day assays (within-study validation) carried out during the analysis of plasma samples collected from healthy volunteers after single-dose treatment with atenolol+chlorthalidone tablets (pharmaceutical preparations containing 100+25 mg and 50+12.5 mg of the two drugs, respectively).     

The use of hexacyanoferrates in different forms to reduce radiocaesium contamination of animal products in Russia

Hexacyanoferrates have been identified as highly effective radiocaesium binders which effectively reduce radiocaesium uptake and transfer to milk and meat. In Russia a hexacyanoferrate called ferrocyn has been produced for use as a countermeasure. In 1989-1992, experiments were undertaken in Russia to study the effectiveness of four different ferrocyn materials as 137Cs binders, their potential toxicity, effect on production rates of cow milk, effect on animal health and ease of implementation in routine agricultural practice. Four different ferrocyn delivery forms have been used: 98% pure powder, sustained release rumen boli (15% ferrocyn), salt licks (10% ferrocyn) and sawdust with 10% ferrocyn adsorbed (bifege). In initial experiments with different cows, sheep and pigs these four ferrocyn materials were effective in reducing radiocaesium transfer to animal products. Daily administration of ferrocyn powder at a rate of 3-5 g per cow reduced 137Cs transfer by up to 90% in milk. One single administration of three boli per cow (containing 30 g ferrocyn per boli) reduced 137Cs transfer by 50-75% for a period of 2 months. Salt licks containing 10% ferrocyn (0.22 kg ferrocyn per 2.2 kg briquette provided once) reduced transfer of 137Cs up to twofold for up to 10 days whilst bifege, given at a rate of 30-60 g day-1 (3-6 g day-1 ferrocyn), reduced 137Cs transfer by 90-95%. However, large-scale application of these ferrocyn materials on collective and private farms in agricultural trials in 1994 resulted in a lower effectiveness. Therefore, in 1996 a comparative assessment of the application of the four ferrocyn forms was made under carefully controlled conditions. The results fully validated the previous experimental data, and showed the importance of meeting recommended procedures for treatment, particularly when hexacyanoferrates are administered on a day-to-day basis.     

Inadequate literacy is a barrier to asthma knowledge and self-care

STUDY OBJECTIVES: To determine the relationship of literacy to asthma knowledge and ability to use a metered-dose inhaler (MDI) among patients with asthma. DESIGN: Cross-sectional survey. SETTING: Emergency department and asthma clinic at an urban public hospital. PATIENTS: Convenience sample of 273 patients presenting to the emergency department for an asthma exacerbation and 210 patients presenting to a specialized asthma clinic for routine care. INTERVENTIONS: Measurement of literacy with the Rapid Estimate of Adult Literacy in Medicine, asthma knowledge (20 question oral test), and demonstration of MDI technique (six-item assessment). MEASUREMENTS AND RESULTS: Only 27% of patients read at the high-school level, although two thirds reported being high-school graduates; 33% read at the seventh- to eighth-grade level, 27% at the fourth- to sixth-grade level, and 13% at or below the third-grade level. Mean asthma knowledge scores (+/-SD) were directly related to reading levels: 15.1+/-2.5, 13.9+/-2.5, 13.4+/-2.8, 11.9+/-2.5, respectively (p < 0.01). Patient reading level was the strongest predictor of asthma knowledge score in multivariate analysis. Poor MDI technique (< or =3 correct steps) was found in 89% of patients reading at less than the third-grade level compared with 48% of patients reading at the high-school level. In multivariate regression analyses, reading level was the strongest predictor of MDI technique. CONCLUSIONS: Inadequate literacy was common and strongly correlated with poorer knowledge of asthma and improper MDI use.     

Activation of factor VII during alimentary lipemia occurs in healthy adults and patients with congenital factor XII or factor XI deficiency, but not in patients with factor IX deficiency

Factor VII activity (FVIIc), a risk marker for coronary heart disease, is increased during postprandial lipemia. Factor VII activation accompanies lipolysis of triglyceride-rich lipoproteins, but the nature of this association and whether it is causal remain uncertain. To explore this issue, four patients with homozygous factor XII deficiency, four with complete factor XI deficiency, six with factor IX deficiency, and their respective age- and sex-matched controls were given two isocaloric dietary regimens, one providing on average 136 g fat and the other 19 g fat. Blood was taken before breakfast, immediately before lunch at 195 minutes, and at completion of the study at 390 minutes. All samples for each subject and matched control were assayed as one batch for FVIIc, activated factor VII, and factor VII antigen (FVIIag). Activation of factor VII was observed with the high-fat regimen but not with the low-fat regimen in all controls, factor XII-deficient patients, and factor XI-deficient patients. No factor VII activation was observed during either regimen in factor IX-deficient patients, but a normal postprandial responsiveness of factor VII to dietary fat was restored in one patient who replicated the study after factor IX therapy. Plasma FVIIag was not altered postprandially in either regimen in any group of patients or controls. Factor IX apparently plays an obligatory role in the postprandial activation of factor VII, although the mechanism remains to be determined.     

A comparative study of the thermal stability of phenylalanyl tRNA synthetase from Escherichia coli MRE-600 and Thermus thermophilus HB 8

The thermal stability of phenylalanyl-tRNA-synthetase (PTS) from E. coli and T.thermophilus HB 8 was studied in solution at various conditions by scanning microcalorimetry. It has been shown that the value of heating rate, concentration of the enzyme and Mg2+ ions in the solution affects the parameters of thermal denaturation of both enzymes. The higher thermal stability of PTS from T. thermophilus was observed as well as the independence of its properties upon broad variations of experimental conditions. The role of thermostability of the enzymes are discussed with respect to the biological properties of E. coli and T.thermophilus.