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A method for estimating sulfhydryl groups by laboratory titrator T-201 is proposed. It consists in culonometric titration of SH-groups by silver ions. The method is highly sensitive and reproducible.
Allergenic potency assessed by the 50% RAST-inhibition endpoint titration was compared with enzyme titration for the analysis of commercial pollen extracts. Six different extracts of short ragweed, orchard grass and perennial rye grass were examined. A good correlation (p < 0.01) was found between the acid phosphatase titers and allergenic potency of short ragweed and perennial rye grass extracts. A similar correlation (p < 0.01) was observed between leucin amino peptidase titers and allergenic potency of short ragweed extracts. No correlation could be found with orchard grass extracts. These findings suggest that enzymatic activity may provide a method for the control of certain commercial pollen extracts.
We describe a procedure for the determination of hydrochloric acid concentration in gastric juice by means of a conductometric titration of the 50-fold diluted sample with an aqueous ammonia solution. The conductometric method of endpoint indication leads to a definite location of the equivalence point in the titration of hydrochloric acid. The proposed method is simple and accurate and shows a good correlation with an accepted method for the measurement of gastric acidity.
Silverwater virus (SIL) was titrated in vivo and in vitro. Using a microculture technique, TCID50 was found to be much more sensitive than both the plaque forming assay and the LD50. The latter was the least sensitive mode of titration of Silverwater virus. The addition of DEAE dextran and trypsin to the viral inoculum increased the number and diameter of the plaques.
Recent literature on electroconvulsive therapy (ECT) emphasizes the use of moderately suprathreshold electrical stimuli for clinical ECT. Since there is substantial interpatient variability in seizure thresholds, individualization of the ECT stimulus dose has been advocated. Three methods for selecting an electrical stimulus are compared in the present study. The methods compared include empirical titration and dose predictive methods described by two different ECT equipment instruction manuals. The use of ECT stimulus titration was found to be a more consistent means of selecting an electrical dose than either of the two predictive methods. The implications of these findings for the clinical practice of ECT are discussed.
The goal of the present study was to determine whether the serotonin (5-HT) system is involved in the effects of kappa opioids as measured with the squirrel monkey shock titration procedure. With this procedure, electric shock was delivered to the monkey's tail and scheduled to increase once every 15 sec from 0.01 to 2.0 mA in 30 steps. Monkeys responded under a fixed ratio 5 schedule to determine the level at which shock intensity was maintained. The intensity below which monkeys maintained shock 50% of the time, or the median shock level (MSL), and the rate of responding in the presence of shock (RR) were determined after the administration of saline and all drug combinations. The kappa opioids U50,488 and spiradoline increased MSL and decreased RR in a dose-dependent manner. The effects of U50,488 and spiradoline on both RR and MSL were enhanced in all three monkeys by the 5-HT2 antagonists ketanserin and pirenperone and in one monkey by another 5-HT2 antagonist, LY 53857. The effects of U50,488, but not spiradoline, were enhanced to a lesser degree by the 5-HT1A receptor agonist 8-OH-DPAT. The effects of U50,488 but not altered by the receptor agonist DOI, the 5-HT3 receptor antagonist MDL 72222 or the alpha-1 adrenergic receptor antagonist prazosin. These results suggest that the effects of kappa opioids in the shock titration procedure probably involve serotonergic mechanisms that are modulated via 5-HT2 and, perhaps, 5-HT1A receptors. Moreover, these interactions probably reflect nonspecific decreases in RR rather than alterations in the antinociceptive effects of kappa opioids.
An established method to determine flux control coefficients is the enzyme titration method in which the change in pathway flux upon a change in the enzyme concentration is measured. In this study, the application of this method to a simple reconstituted pathway was investigated by simulated measurements. The pathway was assumed to be in the quasi-steady-state, which is the experimental realization of the mathematical construct 'steady state'. It was shown that flux control coefficients, calculated in a way that mimics their experimental determination, were strongly time dependent. Initially, the calculated flux control coefficient was high for the enzyme adjacent to the reaction monitoring the flux, and the steady-state value was overestimated. Likewise, flux control coefficients were underestimated for enzymes further away from the monitoring reaction. The observed time course of simulated flux control coefficients was shown to reflect the fact that experimental systems are not steady state but quasi-steady-state. For a pathway in the quasi-steady-state, some of the problems with enzyme titration experiments can be overcome by allowing the system to relax for a time interval that is large compared with the turnover time of the pooled pathway intermediates.
Total serum calcium and magnesium may be determined in one thermometric titration, with disodium ethylenediaminetetraacetate as the titrant. A 1-ml serum sample is diluted with 1 ml of tris(hydroxymethyl)aminomethane buffer (pH 8) and titrated at a constant rate with a motorized syringe buret. Results by the thermometric method compared well with those by atomic absorption spectroscopy.
The squirrel monkey titration procedure was used to assess the antinociceptive effects of the novel delta opioid agonist (+/-)-4-(a-R*)-a(2S*,5R*)-4-allyl-2,5-dimethyl-1-piperazinyl)-3-hydroxyb enzyl)- N,N-diethylbenzamide (BW373U86). Under this procedure shock increased every 15 sec from 0.01 to 2.0 mA in 30 steps. Five responses terminated the shock for 15 sec, after which the shock resumed at a lower intensity. The intensity at which the monkeys maintained the shock 50% of the time (median shock level, MSL) was determined. BW373U86 (1.0-30.0 mg/kg i.m.) increased MSL, but these increases were not dose-dependent and lasted only 15 min or less. Doses of BW373U86 that increased MSL often produced tremors and/or convulsions immediately after administration. When 1.0 mg/kg of naltrindole, a delta opioid antagonist, was given in combination with BW373U86, doses of BW373U86 up to 30 mg/kg no longer increased MSL nor did tremors and/or convulsions occur. Doses of BW373U86 (0.01-0.3 mg/kg i.m.) that did not increase MSL when administered alone shifted the dose-effect curve for the mu agonist l-methadone to the left. These shifts were antagonized dose-dependently by naltrindole. In monkeys that were tolerant to morphine, BW373U86 (0.03-0.1 mg/kg i.m.) shifted the morphine dose-effect curve leftward. In addition, BW373U86 altered the effects of the partial opioid agonists, buprenorphine, nalbuphine, butorphanol and levallorphan such that doses of these drugs that did not increase MSL when administered alone, often did so in the presence of BW373U86. Taken together, these findings indicate that BW373U86 has a delta agonist profile in the squirrel monkey; however, its antinociceptive effects in the shock titration procedure may be due to its toxic effects.
BACKGROUND: A simple and rapid assay system for the detection of human T cell leukemia virus type I (HTLV-I) binding cells was developed to assess the virus specific receptor and titrate the antibodies to block the virus binding. EXPERIMENTAL DESIGN: Cells (5 x 10(5)) were incubated with 100 microliters of the concentrated HTLV-I at 37 degrees C for 1 hour. After washing, the cells were reacted with anti-HTLV-I envelope monoclonal antibody (rat) for 30 minutes on ice and then stained with fluorescein-isothiocyanate-conjugated anti-rat immunoglobulin. The stained samples were analyzed on FACScan. Antibody-titration of the virus-binding inhibition was carried out by pretreatment of the virus with serially diluted sera. RESULTS: The specificity of the virus-binding was shown by dose-response relationship, kinetics of the binding, and temperature dependency. HTLV-I was absorbed onto a wide range of human cell lines and peripheral blood lymphocytes at various levels. Antibodies to inhibit the virus-binding were also quantitatively detected in sera from HTLV-I infected individuals, including asymptomatic carriers and patients with adult T cell leukemia or HTLV-I-associated myelopathy, but not from healthy seronegative controls. CONCLUSIONS: This assay system would be useful in screening the virus-specific receptor and the neutralizing antibodies to HTLV-I. Thus, the assay could be applied to further studies on HTLV-I-related diseases.
The best conditions are selected for amperometric titration of erythrocyte membranes SH-groups by means of silver ions. On the basis of the experimental data, two types of SH-groups according to their capacity of binding Ag+ are found in the membranes. Dynamics of transition of SH-groups of types I into SH-groups of type II is studied. The chemical model explaining the presence of two bends in the course of the membranes amperometric titration curve is created. Calculations were made for the products of R-S-Ag solubility for SH-groups of type I and SH-groups of types I and II as well as for entropy and heat of formation of silver compounds with the membranes sulphydryl groups.
Titrated gamma-globulin, which the authors used in the preoperative management of patients with acquired valvular disease and minimum rheumatism activity, contained high titers of antibodies against the antigens of streptococcus (streptokinase, streptohyaluronidase, streptolysin 0) and Coxsackie A13, A18, and B3 viruses. Its use in combination with antirheumatic agents did not cause any complications and made it possible to reduce the activity of the process in most cases with grade I and after repeated courses also in grade II activity of rheumatism. Treatment with titrated gamma-globulin stimulates the organism's defence forces (factors of nonspecific immunity: complement, lysozyme, interferon) and leads to diminution of the isolation of cardiotropic viruses from blood and bioptic material of the atria of patients.
Two antigen-capture enzyme-linked immunosorbent assays (polyclonal and monoclonal AC-ELISAs) were developed and evaluated for titration of infectious bursal disease viruses (IBDVs) propagated in different host systems, namely BGM-70 continuous cell line, primary chicken embryo fibroblast cells, and chicken bursa of Fabricius. The polyclonal system was more sensitive (P < 0.05) than the monoclonal system but both were specific as indicated by the negative results obtained with three non-IBDVs. The results revealed that the conventional systems used for titration of IBDVs (cell cultures and embryonating chicken eggs) were more sensitive than the polyclonal AC-ELISA.
Proteinuria may be involved in the final common pathway of progressive renal function loss. If so, intervention treatment that reduces proteinuria might prevent or retard long-term renal function loss. In renal patients and in experimental renal disease the severity of proteinuria is associated with the rate of long-term renal function loss. Several large trials on the prevention of long-term renal function loss by antihypertensive treatment with ACE inhibitors (ACEi), were recently completed in diabetic and in non-diabetic renal disease. In those studies long-term renal function loss could indeed be retarded by ACEi; these ACEi regimens were associated with a more effective reduction of proteinuria than control regimens. In studies with a single treatment regimen (drug treatment or a protein restricted diet) a more effective reduction of proteinuria is associated with a more favourable long-term course of renal function as well. As reduction of proteinuria is mostly associated with a lower blood pressure (BP) the respective contributions of the fall in BP and in proteinuria are hard to dissect. Remarkably, however, the efficacy of the reduction of proteinuria (but not of BP) at onset of antihypertensive treatment is predictive of long-term renal outcome. Albeit consistent with a causal role of proteinuria reduction in renoprotection these data cannot distinguish between proteinuria as a marker or a mediator of renal damage. In view of the consistent association of antiproteinuric efficacy with long-term renal outcome we suggest that it would be worthwhile to attempt to improve long-term renoprotection by a strategy aimed at enhancing antiproteinuric efficacy. This approach is feasible as antiproteinuric efficacy of ACEi can be enhanced in several ways, ie, by dietary sodium and protein restriction and by adding a diuretic or indomethacin. Such a strategy would require that titration for adequate BP control is followed by titration for a maximal antiproteinuric effect. If this treatment strategy would improve long-term renal outcome, it would not only be a step forward in the clinical treatment of chronic renal failure, but it would also provide compelling evidence for a causal role of proteinuria in the progression of renal disease.
In order to establish a simple method to titrate antitoxin antibodies in the antivenom sera, we suggest a new immunochemical technic. To a constant dose of a pure toxin extracted from a snake venom, increasing volumes of specific immune serum are added so as an excess of toxin remains free. This residual toxin is titrated by electroimmunodiffusion technic according to Laurell or by radial immunodiffusion method of Mancini. The experimental results in vitro are compared to those obtained in vivo. If a given volume of immune serum neutralize X microgram of toxin by the in vivo technic it will combine in vitro to 2 X or 1.2 X of the same toxin, respectively if we refer to electroimmunodiffusion or to radial immunodiffusion technic results.
Solution pH is a determinant parameter on protein function and stability, and its inclusion in molecular dynamics simulations is attractive for studies at the molecular level. Current molecular dynamics simulations can consider pH only in a very limited way, through a somewhat arbitrary choice of a set of fixed charges on the titrable sites. Conversely, continuum electrostatic methods that explicitly treat pH effects assume a single protein conformation whose choice is not clearly defined. In this paper we describe a general method that combines both titration and conformational freedom. The method is based on a potential of mean force for implicit titration and combines both usual molecular dynamics and pH-dependent calculations based on continuum methods. A simple implementation of the method, using a mean field approximation, is presented and applied to the bovine pancreatic trypsin inhibitor. We believe that this constant-pH molecular dynamics method, by correctly sampling both charges and conformation, can become a valuable help in the understanding of the dependence of protein function and stability on pH.
Most patients with bilateral Meniere's disease treated with streptomycin sulfate titration therapy and followed over the long term have either improvement or relief of their disabling vertigo. Although persistent post-treatment disequilibrium occurs not infrequently, it is mild in the most patients. Stabilization or improvement of hearing occurs in most patients, and the authors have not encountered hearing loss to date as a result of streptomycin sulfate titration therapy. There seems to be considerable separation between the vestibulotoxic and cochleotoxic doses of streptomycin sulfate intramuscularly when administered in humans.
Oven methods for determining moisture (volatiles) in forages and other animal feeds are empirical. The moisture concentration obtained depends upon the time and temperature the sample was dried and is influenced by the presence of other volatiles than water. A validated reference method to measure water in forages and animal feeds could be used to evaluate the appropriateness of oven methods for various types of animal feeds and forages. Karl Fischer titration is a well-established method for determining water. However, thorough extraction of water from forages and feeds is a challenge because they often contain cellular structures that release water slowly. Water was successfully extracted into methanol-formamide (50 + 50) by high-speed homogenization and then titrated directly at 50 degrees C with a one-component Karl Fischer reagent based on imidazole. The method is described in detail, results of day-to-day repeatability and laboratory-to-laboratory reproducibility are reported, and preliminary comparison data between oven methods are provided.