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PubMed1993

Diurnal secretory patterns and tolerance during individually titrated infusions of ranitidine.

BACKGROUND: Gastric acid secretion in humans shows chronobiological patterns that modify the efficacy of antisecretory agents. The ability of individually titrated ranitidine infusions to overcome circadian patterns of gastric acidity and tolerance during prolonged dosing was assessed. METHODS: Eleven healthy subjects were randomized to receive ranitidine infusions of up to 600 mg/24 hours by a pH feedback-regulated pump before and after 9 days of oral dosing with ranitidine, 300 mg four times daily, beginning either in the evening or in the morning in a cross-over, third party-blinded and placebo-controlled study design. RESULTS: Mean 24-hour intravenous ranitidine doses given to attain the target pH of 4 were greater after 9 days of treatment with oral ranitidine than before in the morning and evening studies (P < 0.01). The median 24-hour pH decreased from 5.1 before to 3.6 after oral dosing in the morning study (P < 0.001) and from 4.4 to 2.8 in the evening study (P < 0.001). Before oral dosing of ranitidine, the time of pH > 4 in the evening and morning fasting periods was 71% and 84%, respectively (P < 0.05). After 9 days oral ranitidine, the respective results were 20% and 53% (P < 0.05). CONCLUSIONS: The reduced responsiveness to ranitidine in the evening and the tolerance to ranitidine with repeated dosing were not overcome by individually titrated, high intravenous doses of ranitidine.

Administration, Oral↗
PubMed1995

Low-dose, titratable interferon alfa in decompensated liver disease caused by chronic infection with hepatitis B virus.

BACKGROUND & AIMS: Interferon therapy has been associated with a number of severe side effects when administered to patients with decompensated cirrhosis caused by chronic hepatitis B. The safety and potential efficacy of a low-dose, titratable regimen of interferon alfa-2b in patients with decompensated liver disease caused by chronic hepatitis B virus infection were studied. METHODS: Twenty-six patients were treated at five medical centers. Five patients had Child's class A status, 15 had Child's B status, and 6 had Child's C status. Treatment was continued for 24 weeks whenever possible. Dose adjustments were made according to predefined safety criteria. RESULTS: All patients with Child's A status responded with a sustained loss of serum hepatitis B virus DNA, reduction in aminotransferase activity, and clinical stabilization. Only 5 patients with Child's B (33%) and no patients with Child's C status reached similar end points. The probability of survival was greater in responders than in nonresponders (P = 0.017). Three patients each developed serious infections or greater than twofold increases in serum aminotransferase levels during therapy. CONCLUSIONS: Low-dose, titratable interferon therapy is safer than previously reported regimens. Nonetheless, serious infections were observed relatively frequently, and this therapy should be reserved for individuals with mild to moderate hepatic decompensation, preferably patients with Child's A status.

Adult↗
PubMed1987

The effect of divalent and univalent binding on antibody titration curves in solid-phase ELISA.

This paper describes the influence of antigen coating concentration, epitope density per antigen molecule and anti-immunoglobulin reagents on antibody titration curves in solid-phase ELISA. Based on results obtained with fluorescein as the hapten and monoclonal anti-fluorescein antibody, which were confirmed in another antigen-antibody system, it is concluded that: (a) Antibody titration curves are independent of antigen-coating concentration in a limited range of concentrations only. (b) The complex between one antibody and two epitopes ('divalent binding') is more stable than the complex between one antibody and one epitope ('univalent binding). The ratio between divalent and univalent binding depends on the epitope density per antigen molecule and on the antigen-coating concentration. (c) The prozone phenomenon can be explained by an increased instability of plate bound antibodies due to a shift from divalent to univalent binding. (d) In solid-phase ELISA a correct evaluation of the antiserum specificity can be performed only if it is ascertained that all target antigens are coated under saturating conditions.

Animals↗
PubMed1992

Escherichia coli heat-stable enterotoxin (STa)-biotin conjugates for the titration of STa antisera by an enzyme-linked immunosorbent assay.

The development of a new approach to the diagnosis of infectious diarrhoea, caused by Escherichia coli heat-stable enterotoxin (ST), was preceded by a preliminary study. The purpose of the latter was to establish whether three preparations of ST produced by a human isolate of enterotoxigenic E. coli (STa), obtained at different steps of the purification procedure (involving Amberlite XAD2 resin chromatography (P3), a gel filtration chromatography on a Biogel P4 (P2) or a disc-gel electrophoresis (P1)), could be employed to titrate antisera to STa using an ST-biotin enzyme-linked immunosorbent assay (ELISA). The solid-phase STa was obtained by first coupling the toxin to biotinyl-N-hydroxysuccinimide and then binding this conjugate to avidin adsorbed to flat-bottomed polystyrene microtitre plates. Using these reagents, the assay conditions were examined. Checkerboard tests determined optimal biotin-P3, P2 or P1 toxin conjugate concentrations to be used as the immunosorbent for P3, P2 and P1 antiserum titration. The immunosorbent prepared with STa purified only on Amberlite XAD2 resin was unable to differentiate significantly between P3, P2 or P1 antisera. Immunosorbent prepared with P2 or P1 detected widely differing titres between the three antisera and gave more sensitive results. Only small but questionable differences were observed between P2 and P1 toxin preparations.

Animals↗
PubMed1986

Tifluadom's effects under electric shock titration and tail-immersion procedures in squirrel monkeys.

The analgesic efficacy of the kappa-opioid benzodiazepine, tifluadom, was examined in squirrel monkeys using electric shock titration and tail-immersion procedures. Tifluadom produced dose-dependent increases in the shock intensity that maintained responding under the shock titration schedule without substantially decreasing response rates. Tifluadom also increased the latency of tail-withdrawal from 55 degrees C water. Naloxone attenuated tifluadom's effects under both procedures. These results extend previous reports of tifluadom's analgesic characteristics and suggest that the tail-immersion procedure is a useful analgesic assay in squirrel monkeys.

Analgesia↗
PubMed1983

Effects of clonidine on operant behavior and electric shock titration in the squirrel monkey: effects of alpha 2-adrenoreceptor antagonism.

Squirrel monkeys were studied under fixed-interval schedules in which responding was maintained either by food presentation or by termination of stimuli correlated with electric shock delivery, or under a schedule in which responses decreased the intensity of electric shock, which periodically increased in intensity (shock titration). Clonidine-HCl (0.01-1.7 mg/kg) decreased responding under the fixed-interval schedules and increased the intensity at which shock was maintained under the titration schedule. Pretreatment with the alpha 2-adrenoreceptor antagonist yohimbine-HCl (0.1-1.0 mg/kg) resulted in a 3-10-fold shift to the right in the clonidine dose-response curve. Similar antagonism was seen after pretreatment with small doses of the less selective alpha-adrenoreceptor antagonist, tolazoline-HCl (3.0, 5.6 mg/kg), but not with the alpha 1 antagonist prazosin (0.1, 0.3 mg/kg). The effects of clonidine on behavior under these conditions were apparently due to actions at alpha 2-adrenoreceptor sites.

Adrenergic alpha-Antagonists↗
PubMed1981

Non-bicarbonate buffering of ascites tumor cells in the rat as titrated by strong acids.

Buffer equations originally derived for blood were applied to the in vitro system "ascites tumor cells and ascitic fluid'. Non-bicarbonate buffer values beta(mmol x pH(-1) x 1(-1)) were determined experimentally by titration of ascites, cell free ascitic fluid and cell homogenates with NaOH in the absence of CO2. beta in ascitic fluid, in native ascites and in cell homogenate were beta (F) = 5.9, beta (A) = 16.4 and beta (H) = 24.9, respectively. Intracellular beta value beta (C) calculated from buffer equations depended on the ratio delta pHi/delta pHe. With delta pHi/delta pHe = 1, beta (C) was 38.8. The difference between beta (C) and beta (H) could be partly accounted for by differences in the cellular protein content. With delta pHi/delta pHe = 0.62 as in the presence of CO2, beta (C) becomes 62.6. In the presence of CO2 (see Albers et al., 1981), the extracellular buffering was higher and the intracellular buffering lower than the values titrated in the absence of CO2. It is concluded that in the presence of CO2 and/or HCO3- the apparent intracellular buffering of intact tumor cells results from non-bicarbonate buffering by cell proteins as well as from active or passive ionic exchanges with the extracellular fluid.

Acid-Base Equilibrium↗
PubMed1995

Tight binding affinities determined from thermodynamic linkage to protons by titration calorimetry.

A general titration calorimetry method is described that can be used to determine the affinity of tight binding interactions with proteins. The method is based on the thermodynamic linkage between ligand binding and coupled protonation reactions. The protons linked to a given ligand-binding reaction are measured by titration calorimetry, and integration of the resulting data set yields the pH dependence of the binding affinity based on thermodynamic relationships developed elsewhere. When the pH dependence of the binding affinity is combined with the absolute affinity determined independently at a pH at which the affinity can be conveniently measured, the absolute binding affinity over the entire pH range is determined. The method is well suited for determining high-affinity binding interactions of protein antigens with antibodies, but is applicable to any macromolecular ligand-binding reaction that is coupled to protonation.

Antigen-Antibody Reactions↗
PubMed1979

Free-operant and auto-titration brain self-stimulation procedures in the rat: a comparison of drug effects.

Rats were implanted with bipolar stimulating electrodes aimed at the medial forebrain bundle of the lateral hypothalamus, and trained to press a lever in one of two different procedures in order to receive electrical stimulation through the electrodes. In a free-operant procedure, each response produced a 200 msec train of electric pulses at a suprathreshold current, the intensity of which remained constant throughout the session. In an auto-titration procedure, each response produced an electrical stimulus which was initially set at a suprathreshold intensity. Every 15th response reduced the stimulation current by 3 muA. The animal could reset the current to its initial intensity at any time by pressing a second lever in the test chamber. The average current at which the animal pressed the reset lever was defined as the reinforcement threshold. Dose-response functions were determined for d- and l-amphetamine, alpha-methyltyrosine, and haloperidol. The reinforcement threshold was decreased by both d- and l-amphetamine, increased by haloperidol, and not changed by alpha-methyltyrosine. These effects on reinforcement threshold were not consistently related to the drug-induced changes in response rate in either procedure. The auto-titration procedure may be useful for distinguishing between drugs which cause nonspecific changes in the rate of ongoing behavior and those which specifically modify the reinforcement efficacy of brain stimulation.

Amphetamine↗
PubMed1987

Naloxone sensitivity in squirrel monkeys under a schedule of shock titration.

In order to explore the conditions under which sensitivity to naloxone develops, naloxone's effects were examined in squirrel monkeys responding under a discrete trial schedule of shock titration. Naloxone's effects were examined prior to and following a chronic naloxone regimen in which 10 mg/kg of naloxone was administered daily for at least 21 days. The level at which monkeys maintained the shock and rates of responding were recorded. Prior to chronic administration, selective doses (1.0, 3.0 mg/kg) of naloxone decreased shock levels slightly. These decreases were accompanied by increases in response rates. Subsequent to chronic administration, shock levels were unaltered; however, rates of responding showed greater increases. These results suggest that, following chronic naloxone exposure, animals responding under a schedule of shock titration became more sensitive to the rate-increasing effects of naloxone, but not to naloxone's effects on shock intensity.

Animals↗
PubMed1993

Titrating matching-to-sample performance in pigeons: effects of diazepam, morphine, and cholinergic agents.

Five adult, male White Carneau pigeons were trained to respond under a titrating matching-to-sample schedule of reinforcement. Under this titration schedule, each trial began with the presentation of a sample stimulus (red or green light) on the center key of a three-key pigeon chamber. Completion of 15 responses on the center key resulted in the termination of the stimulus presentation and the initiation of a delay period. The length of the delay changed as a function of the pigeon's performance. During the first five trials of each session, the delay was fixed at 3 s in length. On the sixth and all subsequent trials, the length of the delay was either increased, did not change, or decreased such that accuracy was maintained at approximately 80%. Following the delay, two of the three pigeon keys were transilluminated with different colored lights (red or green). A single response upon the key transilluminated with the same stimulus color as the sample stimulus resulted in the presentation of food. A response on the key transilluminated with the stimulus color that did not match the sample stimulus resulted in a time-out period. Using this procedure, the effects of two drugs of abuse, diazepam (0.03-3 mg/kg) and morphine (0.03-10 mg/kg), a muscarinic antagonist, scopolamine (0.003-0.3 mg/kg), the quaternary derivative of scopolamine, methylscopolamine (0.003-0.3 mg/kg), a cholinesterase inhibitor, physostigmine (0.003-0.1 mg/kg), and the quaternary derivative of physostigmine, neostigmine (0.003-0.1 mg/kg), were determined. Diazepam decreased matching accuracy such that a decrease in the mean delay value for the session was observed.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1995

Linopirdine does not improve matching performance in the titrating matching-to-sample paradigm.

Linopirdine (DUP 996), a proposed cognitive enhancing agent, was studied in four squirrel monkeys (Saimiri sciureus) and six White Carneau pigeons responding under a titrating matching-to-sample paradigm (TMTS). Briefly, under this titration schedule, each trial began with the presentation of a sample stimulus on the center key of a three-key pigeon or squirrel monkey chamber. Completion of a fixed-ratio on the center key resulted in the termination of the stimulus presentation and the initiation of a delay period. The length of the delay changed as a function of the subject's performance. During the first five trials of each session, the delay was fixed at 3 s in length. On the sixth and all subsequent trials, the length of the delay increased, did not change, or decreased such that accuracy was maintained at approximately 80%. Following the delay, two of the three response keys were transilluminated with different colored lights. A single response on the key transilluminated with the same stimulus as the sample stimulus resulted in the presentation of food. A response on the key transilluminated with the stimulus that did not match the sample stimulus resulted in a timeout. Linopridine was administered in the pigeon (0.001-5.6 mg/kg) and squirrel monkey (0.01-1.0 mg/kg) 15 min before testing. Matching performance was not affected as measured by changes in mean delay values or percent accuracy even at doses that decreased rate of responding. These results suggest that the enhancement in cognitive function previously reported after administration of linopiridine may be limited to specific situations.

Animals↗
PubMed1982

A summary of antibody titration experiments in some animal species treated with ERA vaccine and an inactivated rabies vaccine.

The results of antibody titrations in different animal species vaccinated against rabies are reported. The following points are considered: (1) antibody titration may be used to detect an immunity status in dogs, (2) equines should be vaccinated in infected areas, (3) experiments in progress are comparing ERA vaccine and an inactivated vaccine in bovines, and (4) the vaccination of fallow deer (Dama dama) and moufflons (Ovis ammon musimon) produced results suggesting an extension of the experiment with the purpose of vaccinating wild ruminants whenever possible.

Animals↗
PubMed1981

Thermal stability of proteins analyzed by temperature scanning pH-stat titration.

Based on the fact that pH changes occur during the thermal unfolding of a protein, a pH-stat titrimetric procedure is described for the analysis of thermal stability. In all cases the agreement with other stability measurements was good, including a correlation with activity loss in enzymes. A model for the titration curves, assuming first-order denaturation kinetics, linear temperature increase, and validity of the Arrhenius equation, has been proposed and analyzed. Thus, thermodynamic constants can be calculated from titration curves, or transition temperatures estimation if the Arrhenius constants are known. The equipment consists of a pH-stat, a programmable heating unit, and a temperature measuring/recording system. Analysis can be done quickly and on partially purified solutions, provided the buffer capacity is low, using about 20 mg protein/10 ml sample. The effects of pH, Ca2+ ions, substrate, chemical modification, etc., on thermal stability are conveniently analyzed up to about 90 degrees C.

Calcium↗
PubMed1982

Simultaneous determination of delta G, delta H and delta S by an automatic microcalorimetric titration technique. Application to protein ligand binding.

A methodological study has been made with a syringe titration unit attached to an LKB batch microcalorimeter. The precision and accuracy of the instrument assembly have been evaluated by neutralization reactions and by dilution of sucrose solutions. As an example, heat quantities on the order of 10 mJ accompanying the addition of 10 microliter titrant solution could be determined with an accuracy of better than 1%. A stepwise titration procedure was used to characterize the binding of indole-3-propionic acid to alpha-chymotrypsin. The following thermodynamic data were obtained (25 degrees C, acetate buffer, pH 5.80): delta G0=-18.46 +/- 0.17 kJ X mol-1, delta H0=-15.26 +/- 0.20 kJ X mol-1, delta S0 = 10.85 +/- 1.21 JK-1 X mol-1.

Calorimetry↗
PubMed1985

Application of the finite element simulation method to the adiabatic and potentiometric corrections of calorimetric titration data.

A general numerical analysis procedure is described which has been applied to an automated differential pH-thermal titration apparatus operated isoperibolically to obtain thermal corrections for heat loss. It is based on the Direct Byte (D-B) finite element computer simulation technique (FEST) applied to the heat conduction behavior of the instrument with time. Thermal constants of the numerical model are determined, and the results of the correction for titration data obtained from acid-base runs show that a constant upper baseline is achieved using this technique for both fast and slow reactions to an accuracy of 2%. The method is equally valid for endothermic and exothermic reactions.

Calorimetry↗
PubMed1993

New reagents for spectral titration of active site of chymotrypsin.

The reaction of alpha-chymotrypsin (ChT) with 2-phenyl-(E)-[4-(E)-cinnamylidene]oxazolin-5-one (E-PCO) or 2-phenyl-(Z)-[4-(E)-cinnamylidene]oxazolin-5-one (Z-PCO) at pH 4.5 and 25 degrees C led to the quantitative accumulation of the corresponding acyl-ChT intermediate. Very large changes in molar extinction coefficient were observed in the visible region during the conversion of the oxazolinones to the acyl-ChT intermediates. This afforded a reliable method for the spectral titration of ChT in the visible region. Both E-PCO and Z-PCO were stable under these conditions and did not cause any complication in the titration of the active site.

Chymotrypsin↗
PubMed1994

Infectivity titration of the fast-replicating and cytopathic hepatitis A virus strain HM175A.2 by an in situ enzyme immunoassay.

A simple, rapid and objective infectivity assay based on an in situ enzyme immunoassay (EIA) was developed for the fast-growing and cytopathic cell culture-adapted hepatitis A virus (HAV) strain HM175A.2. Infectivity titration by EIA correlated well with titration by cytopathic effects. The reliability of this assay was demonstrated by close agreement in virus infectivity titers among different assays of the same virus aliquot and between assays of different virus aliquots. HAV infected cell cultures after fixation could be stored for up to 1 week before testing without decline in virus titer.

Antigens, Viral↗