Association between HbA1c and the development of cystic fibrosis-related diabetes.

AIMS: To examine HbA1c as a predictor of risk for future development of cystic fibrosis-related diabetes and to assess the association with the development of retinopathy in people with cystic fibrosis-related diabetes. METHODS: A 7-year retrospective longitudinal study was conducted in 50 adults with cystic fibrosis, comparing oral glucose tolerance test results with HbA1c values in predicting the development of cystic fibrosis-related diabetes. Retinal screening data were also compared with HbA1c measurements to assess microvascular outcome. RESULTS: An HbA1c value ≥37 mmol/mol (5.5%; hazard ratio 3.49, CI 1.5-8.1) was significantly associated with the development of dysglycaemia, as defined by the oral glucose tolerance test over a 7-year period. Severity of diabetic retinopathy was associated with a higher HbA1c and longer duration of cystic fibrosis-related diabetes. CONCLUSION: There is a link between HbA1c level and the future development of dysglycaemia in cystic fibrosis based on oral glucose tolerance test, as well as microvascular outcomes. Although current guidance does not advocate the use of HbA1c as a diagnostic tool in cystic fibrosis-related diabetes, it may be of clinical use in determining individuals at risk of future development of cystic fibrosis-related diabetes.

A mathematical model for analysis of pharmacologically induced changes in the kinetics of cardiac muscle.

A mathematical model of the isometric contraction of cardiac muscle is developed and utilized to characterize the inotropic and lusitropic effects of cardioactive compounds in isolated guinea pig left atria. In contrast to metrics that are based on minima and maxima of an isometric twitch and its derivative function, the entire time course of the twitch is used to quantify the kinetics of the contraction-relaxation cycle. The model relates observed tension to a time-dependent activation function that describes generation of internal force and a coupling function that determines mechanical response to the activation function. The model is structured so that it is suitable for nonlinear curve fitting to observed data. Results obtained using the model for fitting experimental data from tissues treated with different classes of cardioactive compounds agree with more qualitative results presented by other authors. Experiments using the model to fit data over an extended (90 min) time course revealed differences in the kinetic profiles of milrinone and forskolin. Computer simulations that demonstrate the effect of each model parameter on twitch kinetics are presented, and the relationships between the model and other theoretical and empirical models of cardiac muscle are discussed. The mathematical model is useful to enable a more quantitative understanding of the kinetics of cardiac muscle contraction and relaxation and identify compounds that may be selective for inotropic or lusitropic effects.

The cubic ternary complex receptor-occupancy model. III. resurrecting efficacy.

Early work in pharmacology characterized the interaction of receptors and ligands in terms of two parameters, affinity and efficacy, an approach we term the bipartite view. A precise formulation of efficacy only exists for very simple pharmacological models. Here we extend the notion of efficacy to models that incorporate receptor activation and G-protein coupling. Using the cubic ternary complex model, we show that efficacy is not purely a property of the ligand-receptor interaction; it also depends upon the distributional details of the receptor species in the native receptor ensemble. This suggests a distinction between what we call potential efficacy (a vector) and realized efficacy (a scalar). To each receptor species in the native receptor ensemble we assign a part-worth utility; taken together these utilities comprise the potential efficacy vector. Realized efficacy is the expectation of these part-worth utilities with respect to the frequency distribution of receptor species in the native receptor ensemble. In the parlance of statistical decision theory, the binding of a ligand to a receptor ensemble is a random prospect and realized efficacy is the utility of this prospect. We explore the implications that our definition of efficacy has for understanding agonism and in assessing the legitimacy of the bipartite view in pharmacology.

Use of resampling techniques to estimate the variance of parameters in pharmacological assays when experimental protocols preclude independent replication: an example using Schild regressions.

Estimates of variance in pharmacological assays are usually made by repeating the experiment with different tissues. Biological factors, such as the inability to wash a drug from tissue, may preclude the type of replication that is appropriate for the statistics of interest. For example, in Schild regressions, replication is usually done at each concentration of antagonist. In some test systems, replication of dose-response curves is not possible. For example, some persistent agonists cannot be removed from tissues after exposure, while in other systems, rapid desensitization severely alters tissue sensitivity to repeated challenge with agonist. In this paper, we demonstrate how a statistical resampling method, bootstrapping, can be used to derive estimates of the confidence intervals for pA2, pKB, and slope from Schild plots. This method utilizes the speed of the computer to estimate variance by repeatedly resampling the data. The advantage to this method is that it can be used for many different experimental designs. For a data set obtained from a Schild regression of atenolol antagonism of isoproterenol in the guinea pig left atrium, bootstrap estimates of confidence limits were calculated for cases where dose ratios were derived from the same tissue and randomly paired tissues. These estimates showed good agreement with estimates obtained using conventional analytical methods, thus suggesting that this method may be useful in practice.

A pharmacodynamic model to investigate the structure-activity profile of a series of novel opioid analgesics.

A simple mathematical model of analgesia in the rat is developed and utilized to determine quantitative structure-activity relationships for a series of novel 4-anilidopiperidine opioids. The compounds tested (selected alkyl carboxyethyl esters attached at the one position of the piperidine ring) were designed for rapid inactivation by blood and tissue esterases. Model parameters included potency and rate constants for loss of pharmacodynamic effect by hydrolysis dependent and independent processes. A significant correlation is observed between duration of pharmacological effect in vivo and the rate constant for hydrolysis in human blood (r = 0.89). In vivo potency shows a moderate correlation with log P2 (r = -0.77). The validity of the model is shown by comparing model-based parameters which characterize potency and duration of effect in vivo with graphically derived parameters. Significant correlations are observed between model and graphically based estimates of potency (r = 0.75) and between model and graphically based estimates of duration of effect (r = 0.70). This model has potential application in studies of other classes of compounds in which hydrolytic cleavage limits duration of pharmacologic effect.

A new method for estimation of agonist dissociation constants (KA): directly fitting the postinactivation concentration-response curve to a nested hyperbolic equation.

The common method for estimating agonist dissociation constants (KA) is the method proposed separately by Furchgott and Mackay. Concentrations of the given agonist producing the same response before ([A]) and after ([A']) irreversible inactivation of a fraction of the receptors (1-q) are described by the equation: 1/[A] = 1/(q*[A']) + (1-q)/(q*KA) and plotted on axes of 1/[A] vs. 1/[A']. The double reciprocal method suffers from the disadvantage that undue weight may be placed on values generated from the smallest observed responses. Our new method of estimating KA and q uses hyperbolic functions to directly fit both concentration-response curves. The control curve is fit to the logistic equation: response = (M* [A]n)/(kn + [A]n); where M is the maximal tissue response, n is the apparent kinetic order of the response at low [A] and k is [A] required for a half-maximal response. The postinactivation concentration-response curve is simultaneously fit to the following equation: response' = M/(((k/(q*KA*[A']))*(KA + [A']* (1-q)))n + 1). This new method was shown to determine KA and q more accurately and precisely than other methods when applied to an artificial data set. In experiments with the rat anococcygeus muscle, the nested hyperbolic method gave estimates of KA with less variance and less range than the double reciprocal method. The nested hyperbolic method was shown to be a valid method of estimating KA and q that has advantages over the other methods.

Theoretical effects of single and multiple transducer receptor coupling proteins on estimates of the relative potency of agonists.

A mathematical model is presented that simulates the steady state kinetics of agonists interacting with a promiscuous receptor. The model system consists of a single receptor that forms a ternary complex with either of two transducer proteins (G proteins). At a given agonist concentration, the concentrations of the two ternary complexes are determined by the relative quantities of the two G proteins and the ratio of the dissociation constants for the two ternary complexes. Accordingly, the potency of an agonist is dependent upon the relative quantities of the G proteins. If receptors are truly promiscuous and if the distribution of G proteins varies with tissue type, then the agonist potency ratio would be tissue dependent as well as receptor dependent. Experimental data from literature studies are reviewed in the context of the promiscuous receptor model, and implications of the model regarding pharmacologic classification of receptors are discussed.

Hepatocyte initiation during continuous administration of diethylnitrosamine and 1,2-sym-dimethylhydrazine.

Hepatocyte initiation, as indexed by growth-selected gamma-glutamyl transferase-positive foci, was measured during continuous exposure to diethylnitrosamine (DEN) at concentrations used in previous DEN bioassay, DNA adduct, and cell replication studies. Hepatocyte initiation increased in a dose- and lobe-specific manner. The efficiency of DEN as an initiating agent was not affected by DEN dose rate over concentrations ranging from 4 to 40 ppm. In a similar experiment the initiating abilities of 1,2-sym-dimethylhydrazine and DEN were compared under continuous exposure regimens. Foci induction increased in a lobe- and time-dependent manner. When left and median lobes were compared, the initiating ability of the two compounds correlated with their carcinogenicity. However, when corresponding anterior lobes were compared, no such correlation was observed.

Binding of formaldehyde to human and rat nasal mucus and bovine serum albumin.

The function of the nasal mucociliary apparatus, an important airway defense mechanism, is inhibited by inhaled formaldehyde. Nasal mucus, which contains significant concentrations of glycoprotein and soluble proteins, is an integral component of this system. This investigation addresses some reactions of formaldehyde with human and rat mucus in vitro in comparison with a model protein, bovine serum albumin. [14C]Formaldehyde was incubated with reconstituted preparations of human and rat nasal mucus or bovine serum albumin. Formaldehyde adducts, stabilized by sodium cyanoborohydride reduction to methylamines, were separated by Sepharose 2B gel filtration. [14C]Formaldehyde bound exclusively to one component of nasal mucus which had an elution volume identical to that of albumin. There was no detectable binding to the large molecular weight glycoproteins. The time course of reaction of formaldehyde to free amino groups was then measured using the fluorescamine technique. Formaldehyde binding was characterized by an initial fast phase (less than 2 min) followed by a slower phase which appeared to approach equilibrium (greater than 60 min). The rate of binding to human and rat nasal mucus was similar to albumin. Irreversible binding of formaldehyde to albumin was insignificant within the first 60 min indicating the reversibility of binding during this time. These data indicate that within the first 60 min, formaldehyde reacts rapidly and reversibly with nasal mucus and that it binds primarily to one component of nasal mucus. Gel filtration analysis suggests this component may be albumin although other low molecular weight proteins cannot be ruled out.

Molecular dosimetry of O4-ethyldeoxythymidine in rats continuously exposed to diethylnitrosamine.

There is considerable interest in incorporating mechanistically based biological data into the process of quantitative risk assessment. Presently, no adequate data bases for internal dosimeters, such as DNA adducts, exist for humans or experimental animals. Therefore, the major promutagenic ethyl adduct, O4-ethyldeoxythymidine (O4-EtdT), has been quantitated in liver DNA after continuous exposure of rats to drinking water containing 0.4, 1, 4, 10, 40, or 100 ppm diethylnitrosamine (DEN) for 1, 4, 7, 14, 28, 49, or 70 days. The rate of O4-EtdT accumulation was modeled as the difference between a DEN-dependent rate of formation and an O4-EtdT concentration-dependent rate of loss. In general, O4-EtdT concentrations increased rapidly during the first 7 days of exposure and by 7-28 days O4-EtdT had accumulated to apparent steady-state concentrations that were DEN concentration-dependent over the entire dose range. The concentration of the adduct increased with DEN concentration over the entire dose range for exposures of 28 days or less and for doses of 0.4 to 40 ppm DEN the adduct level increased with DEN concentration for exposures of 70 days or less. Although the dose response of O4-EtdT was relatively linear, with increasing DEN concentration a trend toward a less than linear relationship was observed. This suggests that there was a lower efficiency of formation and/or greater loss of O4-EtdT during high-dose exposures. This study provides a data base that can be used to qualitatively and quantitatively examine the relationship between external dose and O4-EtdT over a DEN dose range covering several orders of magnitude.

New methods for comparing the biological efficiency of alternate nutrient sources.

The objective of this study is to propose new methods for the determination of biological efficiency (the ability of a nutrient to produce a response) and for comparison of the efficiencies of alternate nutrient sources. The proposed methods are based on a four-parameter kinetic model which describes response as a function of intake. The comparison of the abilities of two proteins (casein and soybean protein concentrate) to promote weight gain in weanling rats is presented as an example; however, the model is also useful for other nutrients (proteins, amino acids, vitamins, minerals, etc.) and other responses (blood enzyme or protein levels, tissue enzyme levels, etc.). Application of the method leads to useful comparisons of nutrient sources as well as information concerning the maximum efficiencies and rates of nutrient utilization from different sources.