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In a perfect world, data could be placed neatly into discrete categories; however, the world we live in is far from ideal. A consistent finding from studying atomic bonds to psychopathy is that variables and people seldom cooperate with our attempts to fit them into neat clusters. The argument for the current work is that ambiguities in group membership may be more informative than we acknowledge, and warrant adoption of "fuzzy" methodologies that can explicitly quantify these uncertainties. Here, we describe fuzzy community detection methods which permit variables to belong to multiple communities by varying degrees rather than restricting their membership to "on" or "off", and contrast them with discrete forms of community detection. In line with recent calls for dimensional approaches to the study of psychopathology, we discuss how the application of these fuzzy methodologies may inform the study of psychological symptoms and subjects. In particular, we discuss how ambiguous symptoms could serve as the bridges between psychopathologies and how ambiguous subjects could exhibit dynamics relevant for treatment.

The vector autoregression (VAR) and its flavors have seen a boom in the behavioral sciences over the past decades due to their relative simplicity for analyzing N = 1 time-series and calls to bring the person back to the scientific study of psychology. The rise in N = 1 analyses is then met by a commensurate rise in methods attempting to reconcile person- and group-specific processes for diagnostic purposes and overall generalizability. While these methods have found great success in the last decade, some questions still remain. Some known challenges inherent to VAR-based approaches include the assumption of equal interval between successive observations, and potential inferential confounds when researchers encounter empirical time intervals vary within or across studies. All of these challenges can be ameliorated by fitting models in continuous-time. Yet, methods for finding nomothetic trends in idiographic processes are mainly developed in the discrete time framework. Building on the work of Molenaar and collaborators on subgrouping individual time series by means of the subgrouped chain graphical VAR (scgVAR) and the subgrouping option in the group iterative multiple model estimation (S-GIMME), we present results from a Monte Carlo study aimed at addressing the implications of identifying subgroups using these discrete-time methods when the underlying processes are continuous in nature, and providing insights to guide future design and analytic decisions.

Recent years have seen the emergence of an "idio-thetic" class of methods to bridge the gap between nomothetic and idiographic inference. These methods describe nomothetic trends in idiographic processes by pooling intraindividual information across individuals to inform group-level inference or vice versa. The current work introduces a novel "idio-thetic" model: the subgrouped chain graphical vector autoregression (scGVAR). The scGVAR is unique in its ability to identify subgroups of individuals who share common dynamic network structures in both lag(1) and contemporaneous effects. Results from Monte Carlo simulations indicate that the scGVAR shows promise over similar approaches when clusters of individuals differ in their contemporaneous dynamics and in showing increased sensitivity in detecting nuanced group differences while keeping Type-I error rates low. In contrast, a competing approach–the Alternating Least Squares VAR (ALS VAR) performs well when groups were separated by larger distances. Further considerations are provided regarding applications of the ALS VAR and scGVAR on real data and the strengths and limitations of both methods.

ObjectiveTo describe the unique case history of a patient with mGluR1 antibodies, with mainly limbic and without cerebellar symptoms.MethodsA 50-year-old woman initially presented with focal seizures with epigastric rising and déjà-vu sensations, next to cognitive complaints, and musical auditory hallucinations. MRI, EEG, and neuronal autoantibody tests were performed.ResultsEEG findings showed slow and sharp activity (sharp waves and sharp-wave–slow-wave complex) in the left temporal lobe. A test for autoantibodies was negative initially. Because of persistent symptoms, serum and CSF were tested 4 years later and found positive for mGluR1 antibodies. Treatment started with monthly IV immunoglobulins and azathioprine that was replaced by mycophenolate mofetil later. Especially cognitive symptoms and hallucinations did not respond well to the treatment. During treatment, mGluR1 antibodies remained present in CSF.DiscussionWhereas cerebellar symptoms are present in 97% of mGluR1-positive cases, our patient presented without ataxia. Therefore, we suggest that the clinical presentation of patients with mGluR1 antibodies is probably more diverse than previously described. Testing for mGluR1 antibodies should be considered in patients with limbic encephalitis and epilepsy, especially when negative for more common antibodies.

This systematic review aimed to examine the magnitude and direction of the associations between prenatal sleep behaviours (i.e. nighttime sleep duration, sleep quality, night awakenings and daytime nap duration) and eating behaviours, physical activity and gestational weight gain. A systematic search was conducted using Medline/PubMed, PsychINFO, CINAHL Complete, ProQuest Dissertations and Thesis AI, and Web of Science to identify studies with at least one sleep measure, and either eating behaviours, physical activity and/or gestational weight gain. In summary, 11 studies met the review criteria and generated 11 total effect size across 10,900 participants. The majority of the studies were conducted after 2010, which highlights the infancy of this research. Overall, the strengths of the effect size were small: sleep-gestational weight gain (effect size = 0.29), sleep-eating behaviours (effect size = 0.13) and sleep-physical activity (effect size = 0.13). The only effect size that emerged as significant was for the pooled sleep behaviours-physical activity association; good sleep behaviours were positively associated with higher levels of physical activity. These findings summarize and provide insight on how sleep behaviours are related to prenatal gestational weight gain, eating behaviours and physical activity by identifying the strength and direction of the associations that have been previously unknown. Results support the rationale for future longitudinal and randomized control trials to examine the effects of sleep behaviours on gestational weight gain, eating behaviours and physical activity over the course of pregnancy.

BACKGROUND: KCNMA1 encodes the α-subunit of the large-conductance Ca(2+)-activated K(+) channel, K(Ca)1.1, and lies within a linkage interval for atrial fibrillation (AF). Insights into the cardiac functions of K(Ca)1.1 are limited, and KCNMA1 has not been investigated as an AF candidate gene. METHODS: The KCNMA1 gene was sequenced in 118 patients with familial AF. The role of K(Ca)1.1 in normal cardiac structure and function was evaluated in humans, mice, zebrafish, and fly. A novel KCNMA1 variant was functionally characterized. RESULTS: A complex KCNMA1 variant was identified in 1 kindred with AF. To evaluate potential disease mechanisms, we first evaluated the distribution of K(Ca)1.1 in normal hearts using immunostaining and immunogold electron microscopy. K(Ca)1.1 was seen throughout the atria and ventricles in humans and mice, with strong expression in the sinus node. In an ex vivo murine sinoatrial node preparation, addition of the K(Ca)1.1 antagonist, paxilline, blunted the increase in beating rate induced by adrenergic receptor stimulation. Knockdown of the K(Ca)1.1 ortholog, kcnma1b, in zebrafish embryos resulted in sinus bradycardia with dilatation and reduced contraction of the atrium and ventricle. Genetic inactivation of the Drosophila K(Ca)1.1 ortholog, slo, systemically or in adult stages, also slowed the heartbeat and produced fibrillatory cardiac contractions. Electrophysiological characterization of slo-deficient flies revealed bursts of action potentials, reflecting increased events of fibrillatory arrhythmias. Flies with cardiac-specific overexpression of the human KCNMA1 mutant also showed increased heart period and bursts of action potentials, similar to the K(Ca)1.1 loss-of-function models. CONCLUSIONS: Our data point to a highly conserved role of K(Ca)1.1 in sinus node function in humans, mice, zebrafish, and fly and suggest that K(Ca)1.1 loss of function may predispose to AF.

Primary graft dysfunction is an important cause of morbidity and mortality after cardiac transplantation. Donor brain stem death (BSD) is a significant contributor to donor heart dysfunction and primary graft dysfunction. There remain substantial gaps in the mechanistic understanding of peritransplant cardiac dysfunction. One of these gaps is cardiac metabolism and metabolic function. The healthy heart is an "omnivore," capable of utilizing multiple sources of nutrients to fuel its enormous energetic demand. When this fails, metabolic inflexibility leads to myocardial dysfunction. Data have hinted at metabolic disturbance in the BSD donor and subsequent heart transplantation; however, there is limited evidence demonstrating specific metabolic or mitochondrial dysfunction. This review will examine the literature surrounding cardiometabolic and mitochondrial function in the BSD donor, organ preservation, and subsequent cardiac transplantation. A more comprehensive understanding of this subject may then help to identify important cardioprotective strategies to improve the number and quality of donor hearts.

We introduce the Boolean network method, a discrete-time dynamical system method, to model the nonlinear dynamics in multivariate systems and to control the system moving to desired state(s). We introduce this method in three steps: (1) inference of the temporal relations between multiple binary variables as Boolean functions, (2) extraction of attractors based on the inferred dynamics and assignment of desirability for each attractor, and (3) design of network control to direct a psychological system toward a desired attractor by identifying how the Boolean network needs to be updated.To demonstrate how the Boolean network can describe and prescribe control for emotion regulation dynamics, we applied this method to data from a study of how children regulate their anger during a frustrating task using bidding and/or distraction (N = 120, T = 480 seconds). Network control strategies were designed to move the child into attractors where anger is OFF. The sample shows heterogeneous emotion regulation dynamics across children in 22 distinct Boolean networks, and heterogeneous control strategies regarding which behavior to perturb and how to perturb it. The Boolean network method provides a novel method to describe nonlinear dynamics in multivariate psychological systems and a control method to guide nonlinear psychological systems toward desired goals.

Mirabegron has been classified as a βRight atrial trabeculae, from non-failing hearts, were paced and contractile force measured at 37°C. Single concentrations of mirabegron were added in the absence or presence of the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (IBMX), βMirabegron significantly increased contractile force in human right atrium (1 μM, 7.6 ± 2.6%, n = 7; 10 μM, 10.2 ± 1.5%, n = 22 compared with (-)-isoprenaline P 0.05). In the presence of IBMX, mirabegron (10 μM) caused a greater contraction. L-748,337 (100 nM) had no effect on the increase in contractile force caused by mirabegron (10 μM). In contrast, mirabegron (10 μM) reduced contractile force in the presence of CGP 20712A, which was not affected by L-748,337 (100 nM) or ICI 118,551 (50 nM). Mirabegron (10 μM) also reduced contractile force in the presence of desipramine or phenoxybenzamine.Mirabegron increases human atrial force through β

A numerical study has been carried out to investigate the boiling flow instability of a reactor thermosyphon system. The numerical model solves the conservation equations of mass, momentum and energy applicable to a two-fluid and three-field steam–water system using a finite difference technique. The computer code MONA was used for this purpose. The code was applied to the thermosyphon system of an EO (ethylene oxide) chemical reactor in which the heat released by a catalytic reaction is carried by boiling water under natural circulation conditions. The steady-state characteristics of the reactor thermosyphon system were predicted using the MONA code and conventional two-phase flow models in order to understand the model applicability for this type of thermosyphon system. The two-fluid model was found to predict the flow closest to the measured value of the plant. The stability behaviour of the thermosyphon system was investigated for a wide range of operating conditions. The effects of power, subcooling, riser length and riser diameter on the boiling flow instability were determined. The system was found to be unstable at higher power conditions which is typical for a Type II instability. However, with an increase in riser diameter, oscillations at low power were observed as well. These are classified as Type I instabilities. Stability maps were predicted for both Type I and Type II instabilities. Methods of improving the stability of the system are discussed.

Cardiac ryanodine receptors (RyR2) from humans, rats, and sheep show differential sensitivity to calcium and magnesium, with regulation of human RyR2 resembling that of sheep more than that of rat., Regulation of the cardiac ryanodine receptor (RyR2) by intracellular Ca2+ and Mg2+ plays a key role in determining cardiac contraction and rhythmicity, but their role in regulating the human RyR2 remains poorly defined. The Ca2+- and Mg2+-dependent regulation of human RyR2 was recorded in artificial lipid bilayers in the presence of 2 mM ATP and compared with that in two commonly used animal models for RyR2 function (rat and sheep). Human RyR2 displayed cytoplasmic Ca2+ activation (Ka = 4 µM) and inhibition by cytoplasmic Mg2+ (Ki = 10 µM at 100 nM Ca2+) that was similar to RyR2 from rat and sheep obtained under the same experimental conditions. However, in the presence of 0.1 mM Ca2+, RyR2s from human were 3.5-fold less sensitive to cytoplasmic Mg2+ inhibition than those from sheep and rat. The Ka values for luminal Ca2+ activation were similar in the three species (35 µM for human, 12 µM for sheep, and 10 µM for rat). From the relationship between open probability and luminal [Ca2+], the peak open probability for the human RyR2 was approximately the same as that for sheep, and both were ∼10-fold greater than that for rat RyR2. Human RyR2 also showed the same sensitivity to luminal Mg2+ as that from sheep, whereas rat RyR2 was 10-fold more sensitive. In all species, modulation of RyR2 gating by luminal Ca2+ and Mg2+ only occurred when cytoplasmic [Ca2+] was

PDE3 and/or PDE4 control ventricular effects of catecholamines in several species but their relative effects in failing human ventricle are unknown. We investigated whether the PDE3-selective inhibitor cilostamide (0.3-1 μM) or PDE4 inhibitor rolipram (1-10 μM) modified the positive inotropic and lusitropic effects of catecholamines in human failing myocardium.Right and left ventricular trabeculae from freshly explanted hearts of 5 non-β-blocker-treated and 15 metoprolol-treated patients with terminal heart failure were paced to contract at 1 Hz. The effects of (-)-noradrenaline, mediated through β₁ adrenoceptors (β₂ adrenoceptors blocked with ICI118551), and (-)-adrenaline, mediated through β₂ adrenoceptors (β₁ adrenoceptors blocked with CGP20712A), were assessed in the absence and presence of PDE inhibitors. Catecholamine potencies were estimated from -logEC₅₀s.Cilostamide did not significantly potentiate the inotropic effects of the catecholamines in non-β-blocker-treated patients. Cilostamide caused greater potentiation (P = 0.037) of the positive inotropic effects of (-)-adrenaline (0.78 ± 0.12 log units) than (-)-noradrenaline (0.47 ± 0.12 log units) in metoprolol-treated patients. Lusitropic effects of the catecholamines were also potentiated by cilostamide. Rolipram did not affect the inotropic and lusitropic potencies of (-)-noradrenaline or (-)-adrenaline on right and left ventricular trabeculae from metoprolol-treated patients.Metoprolol induces a control by PDE3 of ventricular effects mediated through both β₁ and β₂ adrenoceptors, thereby further reducing sympathetic cardiostimulation in patients with terminal heart failure. Concurrent therapy with a PDE3 blocker and metoprolol could conceivably facilitate cardiostimulation evoked by adrenaline through β₂ adrenoceptors. PDE4 does not appear to reduce inotropic and lusitropic effects of catecholamines in failing human ventricle.

It has been proposed that BRL37344, SR58611 and CGP12177 activate β₃-adrenoceptors in human atrium to increase contractility and L-type Ca(2+) current (I(Ca-L)). β₃-adrenoceptor agonists are potentially beneficial for the treatment of a variety of diseases but concomitant cardiostimulation would be potentially harmful. It has also been proposed that (-)-CGP12177 activates the low affinity binding site of the β₁-adrenoceptor in human atrium. We therefore used BRL37344, SR58611 and (-)-CGP12177 with selective β-adrenoceptor subtype antagonists to clarify cardiostimulant β-adrenoceptor subtypes in human atrium.Human right atrium was obtained from patients without heart failure undergoing coronary artery bypass or valve surgery. Cardiomyocytes were prepared to test BRL37344, SR58611 and CGP12177 effects on I(Ca-L). Contractile effects were determined on right atrial trabeculae.BRL37344 increased force which was antagonized by blockade of β₁- and β₂-adrenoceptors but not by blockade of β₃-adrenoceptors with β₃-adrenoceptor-selective L-748,337 (1 µM). The β₃-adrenoceptor agonist SR58611 (1 nM-10 µM) did not affect atrial force. BRL37344 and SR58611 did not increase I(Ca-L) at 37°C, but did at 24°C which was prevented by L-748,337. (-)-CGP12177 increased force and I(Ca-L) at both 24°C and 37°C which was prevented by (-)-bupranolol (1-10 µM), but not L-748,337.We conclude that the inotropic responses to BRL37344 are mediated through β₁- and β₂-adrenoceptors. The inotropic and I(Ca-L) responses to (-)-CGP12177 are mediated through the low affinity site β(1L)-adrenoceptor of the β₁-adrenoceptor. β₃-adrenoceptor-mediated increases in I(Ca-L) are restricted to low temperatures. Human atrial β₃-adrenoceptors do not change contractility and I(Ca-L) at physiological temperature.

In the autoimmune disease myasthenia gravis (MG), autoantibodies against the muscle AChR are mainly produced by both short- and long-lived plasma cells, which are resistant to standard immunosuppressive drugs (i.e. glucocorticoids). A novel therapy to eliminate plasma cells is the proteasome inhibitor bortezomib, which is used to treat patients with multiple myeloma (MM, a plasma cell malignancy). Previously, we demonstrated that bortezomib also reduced autoantibody titers in an animal model of MG (Gomez, A. M. J. Immunol. 2011). The thymus of MG patients is frequently enriched in germinal centers and contains plasma cells that produce autoantibodies in vitro, even after irradiation (which depletes B and T lymphocytes). We studied the in vitro effects of bortezomib in cultured thymus cells from MG patients undergoing therapeutic thymectomy. Treatment with a single dose of bortezomib blocked the production of these pathogenic autoantibodies, reduced the total IgG levels and eliminated plasma cells. Ultrastructural signs of apoptosis were detected in plasma cells as early as 8 h after addition of bortezomib; at 24 h, no plasma cells could be detected. Moreover, we demonstrated that the minimum concentration of bortezomib that eliminates plasma cells in vitro is 60-fold lower than the peak concentration found in MM patients treated with bortezomib, suggesting that low doses might be effective for eliminating plasma cells in patients with antibody-mediated autoimmune diseases.

Ro 03-7894 (0.6 mM) produced a non-parallel shift to the right of dose-response curves to (-)-isoprenaline in K+ depolarized uterine preparations from the guinea-pig. The displacement of the curves was readily reversed by washing. A rightward shift of similar magnitude was also produced by Ro 03-7894 in transmurally stimulated ileal preparations. The relaxant effects of fenoterol in carbachol-contracted guinea-pig tracheal preparations (in the presence of 2 microM atenolol) were not altered by 0.6 mM Ro 03-7894. In the three tissues there was no evidence of a reduction in the maximal inhibitory response to the agonists. In uterine and tracheal preparations, Ro 03-7894 (0.6 mM) depressed contractile responses to exogenous calcium. The depression of responses was enhanced after washout of Ro 03-7894 for 80 min. Contractile responses of ileal preparations to transmural stimulation were also depressed by Ro 03-7894. Concentration-effect curves for the positive inotropic effects of (-)-isoprenaline in guinea-pig left atrial preparations were markedly shifted to the right and the maximum response depressed by 0.6 mM Ro 03-7894. Although the rightward shift of the curves was fully reversed during the 120 min washout period, the maximal responses remained depressed. In similar experiments, Ro 03-7894 produced a washout-resistant depression of inotropic responses to histamine and calcium. The results of radioligand binding studies in left atria using (-)-[125I]-iodocyanopindolol indicated that, when compared to the untreated atria, there was no reduction in the maximal density of binding sites 120 min after washout of 0.6 mM Ro 03-7894. 5 On the basis of the present results it is concluded that Ro 03-7894 induces a non-specific depressant effect on smooth and cardiac muscle preparations during exposure to the drug. This depressant effect persists following washout of the drug. There is no evidence for an irreversible effect of Ro 03-7894 at beta-adrenoceptor sites.

The subtypes of β-adrenoceptor present in left atrial preparations from the guinea-pig and cat have been assessed using both responses obtained in organ bath experiments and radioligand binding studies. From the positive inotropic responses to procaterol, the pKB values for practolol using a variety of agonists, and from displacement of [125 I]cyanopindolol from left atrial membrane homogenates by the selective β1 and β2-adrenoceptor antagonists L643, 717-01J10 and ICI 118, 551, it was concluded that guinea-pig left atria possess only β1-adrenoceptors, whilst cat left atria possess both β1- and β2-adrenoceptor subtypes.

In electrically driven guinea pig left atria, positive inotropic responses to (−)-isoprenaline and the selective β 1 -adrenoceptor agonist RO363 were obtained in the absence and in the presence of the functional antagonists adenosine, carbachol, gallopamil, nifedipine, and Ro 03-7894. Each of the functional antagonists reduced the maximum response to both agonists and produced nonparallel right-ward shifts in the cumulative concentration effect curves. For both agonists, dissociation constants (k a ) were calculated using the equation described by Furchgott (1966) for irreversible antagonism. For RO363, which is a partial agonist with high agonist activity, the equations outlined for functional interaction by Mackay (1981) were also employed to calculate k a values. The k a values obtained by each method were compared with the dissociation constants (K D ) for the two agonists determined from their ability to displace the radioligand (−)-[ 125 l]iodocyano-pindolol from β 1 -adrenoceptors in guinea pig left atrial membrane preparations. The estimates of k A varied substantially from K D values. The K D values were taken as more accurate estimates of the true values for the dissociation constants because a high degree of correlation exists between pK D and pD 2 values for a number of other β-adrenoceptor agonists that behave as partial agonists and between pK D and pK B values for a number of β-adrenoceptor antagonists. Thus it appears that there are serious limitations in the current theory for using functional antagonism as a means of obtaining agonist dissociation constants.

(-)-Isoprenaline and the selective beta 1-adrenoceptor agonist RO363 were tested for their inotropic effects in left atrial (beta 1) and relaxant effects in K+-depolarized uterine (beta 2) preparations from the guinea-pig. The drugs had similar activities as positive inotropic agents but RO363 was approximately 400 times less active than (-)-isoprenaline as a uterine relaxant. RO363 had intrinsic activities of 0.8 and 0.25 ((-)-isoprenaline = 1) in atrial and uterine preparations, respectively. Apparent dissociation constants (KD values) determined from the ability of the agonists to displace (-)-[125I]-iodocyanopindolol ([125I]-CYP) bound to membranes prepared from both tissues were used as a measure of affinity. The [125I]-CYP binding sites possessed the characteristics of homogeneous populations of beta 1-adrenoceptors in atrial and beta 2-adrenoceptors in uterine membrane preparations. The pKD values for (-)-isoprenaline were similar in the two tissues (left atria 6.4, uterus 6.0) whilst for RO363 the atrial value (7.8) was considerably greater than that for the uterus (6.0). The latter value is very similar to the pKB value determined from shifts in (-)-isoprenaline curves produced by RO363 in uterine preparations. Graphical plots of the fraction of receptors occupied vs response were constructed. The relative efficacy of (-)-isoprenaline with respect to RO363 was calculated to be 25 in atrial and 2633 in uterine preparations. The selective beta 1-adrenoceptor stimulant actions of RO363 are a reflection of both its greater affinity and efficacy for beta 1- as opposed to beta 2-adrenoceptor sites. The potent actions of (-)- isoprenaline in both tissues are largely dependent on efficacy.

The affinity and efficacy of a number of sympathomimetic amines structurally related to prenalterol and the selective beta 1-adrenoceptor agonist RO363 were determined using a combination of radioligand binding and organ bath techniques. Affinity of the molecules (pKD) was calculated from their ability to displace the radioligand [125I]iodocyanopindolol ([125I]CYP) from beta-adrenoceptor sites in left atrial (beta 1) and uterine (beta 2) membrane homogenates. These pKD values were used to calculate efficacy from the positive inotropic and uterine relaxant responses elicited by the drugs in organ bath experiments. The drugs studied were either arylethanolamines i.e., (-)-isoprenaline (ISO), p-hydroxyisoprenaline (pOH-ISO), compounds XIV and XVI or aryloxypropanolamine-derivatives, i.e., oxymethylene-isoprenaline (OM-ISO), prenalterol and Compound XI which possessed a p-phenol or catechol ring and an isopropyl or a homoveratryl amine substituent. Only ISO, OM-ISO, pOH-ISO and Compound XVI were active as agonists in both tissue preparations. These drugs were partial agonists which exhibited a wide range of pD2 values and did not display any marked selectivity for either beta-adrenoceptor subtype. Compound XI and prenalterol were inactive as agonists and together with the partial agonists behaved as competitive antagonists to ISO in the two preparations. All drugs tested displaced [125I]CYP from beta-adrenoceptor sites, however, there was also a wide range of potency amongst the drugs. Analysis of the structure-affinity and structure-efficacy relationships indicated that removal of the 3-hydroxyl group from the catechol ring reduces both affinity and efficacy without altering the selectivity of the drug for either beta-adrenoceptor subtype.(ABSTRACT TRUNCATED AT 250 WORDS)

The contribution of affinity and efficacy to the agonistic actions of the naturally occurring catecholamines, (-)-noradrenaline and (-)-adrenaline at β-adrenoceptor sites were assessed in guinea-pig driven left atrial (β1) and K+-depolarized uterine (β2) preparations. The dissociation constants of each agonist, required in these calculations, were calculated using radioligand binding techniques. [125I]Iodocyanopindolol bound to sites in membrane preparations of each tissue which have been shown to represent β1(atria) and β2-(uterus) adrenoceptors. It was found that (-)-noradrenaline was approximately 10-fold more selective for the β1- as opposed to the β2-adrenoceptor in the pharmacological studies. Affinity/efficacy calculations indicated that this selectivity was entirely due to a selective affinity for the β1-adrenoceptor subtype. (-). Noradrenaline, (-)-adrenaline and the reference compound (-)-isoprenaline all had approximately the same efficacy at either β-adrenoceptor subtype.

The KA value of a full agonist (Furchgott, 1966) is usually calculated following the transformation of a rectangular hyperbolic relationship between equiactive concentrations into a linear relationship via a double reciprocal plot. Such a transformation will introduce error into the value of KA unless the pre- and postinactivation dose-response curves are also rectangular hyperbolas. Nonlinear curve fitting computer programs, which have been developed to evaluate radioligand binding data, fit data to rectangular hyperbolas and, thus, can be used to fit concentration-response data for pre- and postinactivation curves. Examination of (-)-isoprenaline concentration-response curves for positive inotropic effects in left atrial preparations from the guinea pig, established before and after treatment with the irreversible antagonist Ro 03-7894, indicated a marked deviation from the theoretical rectangular hyperbola (Hill coefficient greater than 1). This was accounted for by the presence of a threshold effect. The present study reports on the use of the computer program LIGAND to fit a rectangular hyperbola to the pre- and postinactivation curves for an agonist, while simultaneously calculating and allowing for the presence of a threshold phenomenon. Since both curves are fitted to rectangular hyperbolas, the KA values can be calculated with greater precision than that possible from double reciprocal plots constructed from uncorrected concentration-response curves.

The present investigation has assessed the influence of assay buffer (Tris versus Krebs) on the abilities of several beta-adrenoceptor agonists and antagonists to displace the radioligand (-)-[125I]iodocyanopindolol [( 125I]CYP) from beta 1-(left atrial) and beta 2- (uterine) adrenoceptor sites. Saturation studies indicated that the dissociation constant (KD) for [125I]CYP at both beta-receptor sites was approximately 2-fold greater in Krebs as opposed to Tris buffer, and that the maximal density of binding sites (Bmax) in atria (but not uterus) was also reduced 2-fold. In general, the KD values for beta-adrenoceptor agonists were more influenced by the type of buffer used than were KD values for antagonists. Agonist KD values at beta 2-adrenoceptor sites were higher in Krebs than in Tris buffer, while at beta 1-adrenoceptor sites, variable changes resulted. The selective affinity of agonists at beta 1- and beta 2-adrenoceptor sites was therefore markedly influenced by the buffer used and could not be predicted from the established beta 1-/beta 2-adrenoceptor selectivity of the agonists as found in organ bath studies.