Perspective on the relationship between GABAA receptor activity and the apparent potency of an inhibitor

Background : In electrophysiological experiments inhibition of a receptor-channel, such as the GABAA receptor, is measured by co-applying an agonist producing a predefined control response with an inhibitor to calculate the fraction of the control response remaining in the presence of the inhibitor. The properties of the inhibitor are determined by fitting the inhibition concentration-response relationship to the Hill equation to estimate the midpoint (IC50) of the inhibition curve. Objective: We sought to estimate here the sensitivity of the fitted IC50 to the level of activity of the control response. Methods: The inhibition concentration-response relationships were calculated for models with distinct mechanisms of inhibition. In Model I, the inhibitor acts allosterically to stabilize the resting state of the receptor. In Model II, the inhibitor competes with the agonist for a shared binding site. In Model III, the inhibitor stabilizes the desensitized state. Results: The simulations indicate that the fitted IC50 of the inhibition curve is sensitive to the degree of activity of the control response. In Models I and II, the IC50 of inhibition was increased as the probability of being in the active state (PA) of the control response increased. In Model III, the IC50 of inhibition was reduced at higher PA. Conclusions: We infer that the apparent potency of an inhibitor depends on the PA of the control response. While the calculations were carried out using the activation and inhibition properties that are representative of the GABAA receptor, the principles and conclusions apply to a wide variety of receptor-channels.

Approximate Lyapunov-perron Transformations: Computation and Applications to Quasi-periodic Systems

A new technique for the analysis of dynamical equations with quasi-periodic coefficients (so-called quasi-periodic systems) is presented. The technique utilizes Lyapunov-Perron (L-P) transformation to reduce the linear part of a quasi-periodic system into the time-invariant form. A general approach for the construction of L-P transformations in the approximate form is suggested. First, the linear part of a quasi-periodic system is replaced by a periodic system with a 'suitable' large principal period. Then, the state transition matrix of the periodic system is computed in the symbolic form using Floquet theory. Finally, Lyapunov-Floquet theorem is used to compute approximate L-P transformations. A two-frequency quasi-periodic system is studied and transformations are generated for stable, unstable and critical cases. The effectiveness of these transformations is demonstrated by investigating three distinct quasi-periodic systems. They are applied to a forced quasi-periodic Hill equation to generate analytical solutions. It is found that the closeness of the analytical solutions to the exact solutions depends on the principal period of the periodic system. A general approach to obtain the stability bounds on linear quasi-periodic systems with stochastic perturbations is also discussed. Finally, the usefulness of approximate L-P transformations to nonlinear quasi-periodic systems is presented by analyzing a quasi-periodic Hill equation with cubic nonlinearity using time-dependent normal form theory. The closed-form solution generated is found to be in good agreement with the exact solution.

The polyaminoisoprenyl potentiator NV716 revives old disused antibiotics against intracellular forms of infection by Pseudomonas aeruginosa

Active efflux confers intrinsic resistance to multiple antibiotics in Pseudomonas aeruginosa, including old disused molecules. Beside resistance, intracellular survival is another reason for failure to eradicate bacteria with antibiotics. We evaluated the capacity of polyaminoisoprenyl potentiators (designed as efflux pump inhibitors [EPIs]) NV716 and NV731 compared to PAβN to restore the activity of disused antibiotics (doxycycline, chloramphenicol [substrates for efflux], rifampin [not substrate]) in comparison with ciprofloxacin against intracellular P. aeruginosa (strains with variable efflux levels) in THP-1 monocytes exposed during 24h to antibiotics alone (0.003-100x MIC) or combined with EPIs. Pharmacodynamic parameters (apparent static concentrations [Cs]; maximal relative efficacy [Emax]) were calculated using the Hill equation of concentration-response curves. PAβN and NV731 moderately reduced (0-4 doubling dilutions) antibiotic MICs but did not affect their intracellular activity. NV716 markedly reduced (1-16 doubling dilutions) the MIC of all antibiotics (substrates or not for efflux; strains expressing efflux or not); it improved their relative potency and maximal efficacy (lower Cs; more negative Emax) intracellularly. In parallel, NV716 reduced the persister fraction in stationary cultures when combined with ciprofloxacin. In contrast to PAβN and NV731 that act as EPIs against extracellular bacteria only, NV716 can resensitize P. aeruginosa to antibiotics whether substrates or not for efflux, both extracellularly and intracellularly. This suggests a complex mode of action that goes beyond a simple inhibition of efflux and reduces bacterial persistence. NV716 may appear as a useful adjuvant, including to disused antibiotics with low antipseudomonal activity, to improve their activity, including against intracellular P. aeruginosa.

A Technique for Seamless Forecast Construction and Validation from Weather to Monthly Time Scales

Seamless prediction means bridging discrete short-term weather forecasts valid at a specific time and time-averaged forecasts at longer periods. Subseasonal predictions span this time range and must contend with this transition. Seamless forecasts and seamless validation methods go hand-in-hand. Time-averaged forecasts often feature a verification window that widens in time with growing forecast leads. Ideally, a smooth transition across daily to monthly time scales would provide true seamlessness—a generalized approach is presented here to accomplish this. We discuss prior attempts to achieve this transition with individual weighting functions before presenting the two-parameter Hill equation as a general weighting function to blend discrete and time-averaged forecasts, achieving seamlessness. The Hill equation can be tuned to specify the lead time at which the discrete forecast loses dominance to time-averaged forecasts, as well as the swiftness of the transition with lead time. For this application, discrete forecasts are defined at any lead time using a Kronecker delta weighting, and any time-averaged weighting approach can be used at longer leads. Time-averaged weighting functions whose averaging window widens with lead time are used. Example applications are shown for deterministic and ensemble forecasts and validation and a variety of validation metrics, along with sensitivities to parameter choices and a discussion of caveats. This technique aims to counterbalance the natural increase in uncertainty with forecast lead. It is not meant to construct forecasts with the highest skill, but to construct forecasts with the highest utility across time scales from weather to subseasonal in a single seamless product.

Cooperative Binding and Stepwise Encapsulation of Drug Molecules by Sulfonylcalixarene-Based Metal-Organic Supercontainers

The cooperative binding behavior of a face-directed octahedral metal-organic supercontainer featuring one endo cavity and six exo cavities was thoroughly examined in chloroform solution through ultraviolet-visible (UV-Vis) titration technique using two representative drug molecules as the guests. The titration curves and their nonlinear fit to Hill equation strongly suggest the efficient encapsulation of the guest molecules by the synthetic host, which exhibit interesting cooperative and stepwise binding behavior. Based on the control experiments using tetranuclear complex as a reference, it is clear that two equivalents of the guest molecules are initially encapsulated inside the endo cavity, followed by the trapping of six additional equivalents of the drug molecules through six exo cavities (1 eq. per exo cavity), and the remaining guests are entrapped by the external pockets. The results provide an in-depth understanding of the cooperative binding behavior of metal-organic supercontainers, which opens up new opportunities for designing synthetic receptors for truly biomimetic functional applications.

Degree-day models to predict carrot weevil (Coleoptera: Curculionidae) emergence and oviposition in Nova Scotia, Canada

Carrot weevil, Listronotus oregonensis (LeConte) (Coleoptera: Curculionidae), is a pest of carrot (Daucus carota var. sativus Hoffmann; Apiaceae) throughout eastern Canada. Carrot weevil emergence and oviposition were monitored in commercial carrot fields in Nova Scotia. Cumulative degree days were calculated using a base temperature of 7 °C (DD7), and models were developed to predict cumulative emergence and oviposition using nonlinear regression. Cumulative emergence and oviposition were adequately explained as functions of DD7 by a three-parameter sigmoidal Hill equation. Our emergence model predicted initial and peak adult emergence at 35 and 387 DD7, respectively, with oviposition on carrot baits occurring as early as 42 DD7. Models were then validated to evaluate how well they performed. Oviposition on carrot plants began at the fourth true-leaf stage (342 DD7) and continued until eleventh true-leaf stage. Growers using these models can identify their window of opportunity to manage their carrot weevil populations targeting the majority of emerged adults before oviposition begins in the field.