Bridging the gap between non-canonical and canonical Wnt signaling through Vangl2.

Non-canonical Wnt signaling (encompassing Wnt/PCP and WntCa2+) has a dual identity in the literature. One stream of research investigates its role in antagonizing canonical Wnt/ß-catenin signaling in cancer, typically through Ca2+, while the other stream investigates its effect on polarity in development, typically through Vangl2. Rarely do these topics intersect or overlap. What has become clear is that Wnt5a can mobilize intracellular calcium stores to inhibit Wnt/ß-catenin in cancer cells but there is no evidence that Vangl2 is involved in this process. Conversely, Wnt5a can independently activate Vangl2 to affect polarity and migration but the role of calcium in this process is also limited. Further, Vangl2 has also been implicated in inhibiting Wnt/ß-catenin signaling in development. The consensus is that a cell can differentiate between canonical and non-canonical Wnt signaling when presented with a choice, always choosing non-canonical at the expense of canonical Wnt signaling. However, these are rare events in vivo. Given the shared resources between non-canonical and canonical Wnt signaling it is perplexing that there is not more in vivo evidence for cross talk between these two pathways. In this review we discuss the intersection of non-canonical Wnt, with a focus on Wnt/PCP, and Wnt/ß-catenin signaling in an attempt to shed some light on pathways that rarely meet at a crossroads in vivo.

Prevalence of SARS-CoV-2 Infection among Children and Adults in 15 US Communities, 2021

During January-August 2021, the Community Prevalence of SARS-CoV-2 Study used time/location sampling to recruit a cross-sectional, population-based cohort to estimate SARS-CoV-2 seroprevalence and nasal swab sample PCR positivity across 15 US communities. Survey-weighted estimates of SARS-CoV-2 infection and vaccine willingness among participants at each site were compared within demographic groups by using linear regression models with inverse variance weighting. Among 22,284 persons >2 months of age and older, median prevalence of infection (prior, active, or both) was 12.9% across sites and similar across age groups. Within each site, average prevalence of infection was 3 percentage points higher for Black than White persons and average vaccine willingness was 10 percentage points lower for Black than White persons and 7 percentage points lower for Black persons than for persons in other racial groups. The higher prevalence of SARS-CoV-2 infection among groups with lower vaccine willingness highlights the disparate effect of COVID-19 and its complications.

Pharmacologic characterization of atogepant: A potent and selective calcitonin gene-related peptide receptor antagonist.

BACKGROUND: The trigeminal sensory neuropeptide calcitonin gene-related peptide (CGRP) is identified as an essential element in migraine pathogenesis. METHODS: In vitro and in vivo studies evaluated pharmacologic properties of the CGRP receptor antagonist atogepant. Radioligand binding using 125I-CGRP and cyclic adenosine monophosphate (cAMP) accumulation assays were conducted in human embryonic kidney 293 cells to assess affinity, functional potency and selectivity. Atogepant in vivo potency was assessed in the rat nitroglycerine model of facial allodynia and primate capsaicin-induced dermal vasodilation (CIDV) pharmacodynamic model. Cerebrospinal fluid/brain penetration and behavioral effects of chronic dosing and upon withdrawal were evaluated in rats. RESULTS: Atogepant exhibited high human CGRP receptor-binding affinity and potently inhibited human α-CGRP-stimulated cAMP responses. Atogepant exhibited significant affinity for the amylin1 receptor but lacked appreciable affinities for adrenomedullin, calcitonin and other known neurotransmitter receptor targets. Atogepant dose-dependently inhibited facial allodynia in the rat nitroglycerine model and produced significant CIDV inhibition in primates. Brain penetration and behavioral/physical signs during chronic dosing and abrupt withdrawal were minimal in rats. CONCLUSIONS: Atogepant is a competitive antagonist with high affinity, potency and selectivity for the human CGRP receptor. Atogepant demonstrated a potent, concentration-dependent exposure/efficacy relationship between atogepant plasma concentrations and inhibition of CGRP-dependent effects.

Influence of repulsion on entropy scaling and density scaling of monatomic fluids.

Entropy scaling is applied to the shear viscosity, self-diffusion coefficient, and thermal conductivity of simple monatomic fluids. An extensive molecular dynamics simulation series is performed to obtain these transport properties and the residual entropy of three potential model classes with variable repulsive exponents: n, 6 Mie (n = 9, 12, 15, and 18), Buckingham's exponential-six (α = 12, 14, 18, and 30), and Tang-Toennies (αT = 4.051, 4.275, and 4.600). A wide range of liquid and supercritical gas- and liquid-like states is covered with a total of 1120 state points. Comparisons to equations of state, literature data, and transport property correlations are made. Although the absolute transport property values within a given potential model class may strongly depend on the repulsive exponent, it is found that the repulsive steepness plays a negligible role when entropy scaling is applied. Hence, the plus-scaled transport properties of n, 6 Mie, exponential-six, and Tang-Toennies fluids lie basically on one master curve, which closely corresponds with entropy scaling correlations for the Lennard-Jones fluid. This trend is confirmed by literature data of n, 6 Mie, and exponential-six fluids. Furthermore, entropy scaling holds for state points where the Pearson correlation coefficient R is well below 0.9. The condition R > 0.9 for strongly correlating liquids is thus not necessary for the successful application of entropy scaling, pointing out that isomorph theory may be a part of a more general framework that is behind the success of entropy scaling. Density scaling reveals a strong influence of the repulsive exponent on this particular approach.

Entropy scaling of viscosity for molecular models of molten salts.

Entropy scaling relates dynamic and thermodynamic properties by reducing the viscosity to a function of only the residual entropy. Molecular simulations are used to investigate the entropy scaling of the viscosity of three models of sodium chloride and five monovalent salts. Even though the correlation between the potential energy and the virial is weak, entropy scaling applies at liquid densities for all models and salts investigated. At lower densities, entropy scaling breaks down due to the formation of ion pairs and chains. Entropy scaling can be used to develop more extendable correlations for the dynamic properties of molten salts.

Unraveling the Chemosensing Mechanism by the 7-(Diethylamino)coumarin-hemicyanine Hybrid: A Ratiometric Fluorescent Probe for Hydrogen Peroxide.

The hemicyanine hybrid containing the 7-(diethylamino)coumarin (ACou) donor attached to the cationic indolenium (Ind) acceptor through a vinyl linkage (ACou-Ind) represents a classic ratiometric fluorescent probe for detecting nucleophilic analytes, such as cyanide and reactive sulfur species (RSS), through addition reactions that disrupt dye conjugation to turn off red internal charge transfer (ICT) fluorescence and turn on blue coumarin emission. The chemosensing mechanism for RSS detection by ACou-Ind suggested in the literature has now been revised. Our studies demonstrate that thiolates react with ACou-Ind through conjugate addition to afford C4-SR adducts that lack coumarin fluorescence due to photoinduced electron transfer quenching by the electron-rich enamine intermediate. Thus, ACou-Ind serves as a turn-off probe through loss of red ICT fluorescence upon RSS addition. The literature also suggests that blue coumarin emission of thiolate adducts is enhanced in the presence of reactive oxygen species (ROS) due to ROS-mediated cellular changes. Our studies predict that such a scenario is unlikely and that thiolate adducts undergo oxidative deconjugation in the presence of H2O2, the pervasive ROS. Under basic conditions, H2O2 also reacts directly with ACou-Ind to generate intense coumarin fluorescence through an epoxidation process. The relevance of our chemosensing mechanism for ACou-Ind was assessed within live zebrafish, and implications for the utility of ACou-Ind for unraveling the interplay between RSS and ROS are discussed.

Connecting entropy scaling and density scaling.

It is shown that the residual entropy (entropy minus that of the ideal gas at the same temperature and density) is mostly synonymous with the independent variable of density scaling, identifying a direct link between these two approaches. The residual entropy and the effective hardness of interaction (itself a derivative at constant residual entropy) are studied for the Lennard-Jones monomer and dimer as well as a range of rigid molecular models for carbon dioxide. It is observed that the density scaling exponent appears to be related to the two-body interactions in the dilute-gas limit.

Probing the link between residual entropy and viscosity of molecular fluids and model potentials.

This work investigates the link between residual entropy and viscosity based on wide-ranging, highly accurate experimental and simulation data. This link was originally postulated by Rosenfeld in 1977 [Rosenfeld Y (1977) Phys Rev A 15:2545-2549], and it is shown that this scaling results in an approximately monovariate relationship between residual entropy and reduced viscosity for a wide range of molecular fluids [argon, methane, [Formula: see text], [Formula: see text], refrigerant R-134a (1,1,1,2-tetrafluoroethane), refrigerant R-125 (pentafluoroethane), methanol, and water] and a range of model potentials (hard sphere, inverse power, Lennard-Jones, and Weeks-Chandler-Andersen). While the proposed "universal" correlation of Rosenfeld is shown to be far from universal, when used with the appropriate density scaling for molecular fluids, the viscosity of nonassociating molecular fluids can be mapped onto the model potentials. This mapping results in a length scale that is proportional to the cube root of experimentally measurable liquid volume values.

Survey of Data and Models for Refrigerant Mixtures Containing Halogenated Olefins.

We survey existing data for refrigerant blends containing halogenated olefins (hydrofluoroolefins (HFO), hydrochlorofluoroolefins (HCFO) and hydrochloroolefins (HCO)) in the open literature. The data are primarily taken from the NIST SOURCE database and are presented in graphical form to demonstrate the relative coverage of the data. The primary conclusion is that blends containing halogenated olefins remain only sparsely measured in experiments, and some classes of data (e.g., speed-of-sound data) are particularly sparse for blends containing halogenated olefins. The second part of this study compares the thermodynamic models in NIST REFPROP against the experimental datasets and identifies systems (of which there are many) where refitting of the thermodynamic model is required.

Characterization of Ubrogepant: A Potent and Selective Antagonist of the Human Calcitonin Gene‒Related Peptide Receptor.

A growing body of evidence has implicated the calcitonin gene-related peptide (CGRP) receptors in migraine pathophysiology. With the recent approval of monoclonal antibodies targeting CGRP or the CGRP receptor, the inhibition of CGRP-mediated signaling has emerged as a promising approach for preventive treatments of migraine in adults. However, there are no small-molecule anti-CGRP treatments available for treating migraine. The current studies aimed to characterize the pharmacologic properties of ubrogepant, an orally bioavailable, CGRP receptor antagonist for the acute treatment of migraine. In a series of ligand binding assays, ubrogepant exhibited a high binding affinity for native (K i=0.067 nM) and cloned human (K i=0.070 nM) and rhesus CGRP receptors (K i=0.079 nM), with relatively lower affinities for CGRP receptors from rat, mouse, rabbit and dog. In functional assays, ubrogepant potently blocked human α-CGRP stimulated cAMP response (IC50 of 0.08 nM) and exhibited highly selective antagonist activity for the CGRP receptor compared with other members of the human calcitonin receptor family. Furthermore, the in vivo CGRP receptor antagonist activity of ubrogepant was evaluated in a pharmacodynamic model of capsaicin-induced dermal vasodilation (CIDV) in rhesus monkeys and humans. Results demonstrated that ubrogepant produced concentration-dependent inhibition of CIDV with a mean EC50 of 3.2 and 2.6 nM in rhesus monkeys and humans, respectively. Brain penetration studies with ubrogepant in monkeys showed a CSF/plasma ratio of 0.03 and low CGRP receptor occupancy. In summary, ubrogepant is a competitive antagonist with high affinity, potency, and selectivity for the human CGRP receptor. SIGNIFICANCE STATEMENT: Ubrogepant is a potent, selective, orally delivered, small-molecule competitive antagonist of the human calcitonin generelated peptide receptor. In vivo studies using a pharmacodynamic model of capsaicin-induced dermal vasodilation (CIDV) in rhesus monkeys and humans demonstrated that ubrogepant produced concentration-dependent inhibition of CIDV, indicating a predictable pharmacokinetic-pharmacodynamic relationship.

Pharmacological Characterization of the Novel and Selective α7 Nicotinic Acetylcholine Receptor-Positive Allosteric Modulator BNC375.

Treatments for cognitive deficits associated with central nervous system (CNS) disorders such as Alzheimer disease and schizophrenia remain significant unmet medical needs that incur substantial pressure on the health care system. The α7 nicotinic acetylcholine receptor (nAChR) has garnered substantial attention as a target for cognitive deficits based on receptor localization, robust preclinical effects, genetics implicating its involvement in cognitive disorders, and encouraging, albeit mixed, clinical data with α7 nAChR orthosteric agonists. Importantly, previous orthosteric agonists at this receptor suffered from off-target activity, receptor desensitization, and an inverted U-shaped dose-effect curve in preclinical assays that limit their clinical utility. To overcome the challenges with orthosteric agonists, we have identified a novel selective α7 positive allosteric modulator (PAM), BNC375. This compound is selective over related receptors and potentiates acetylcholine-evoked α7 currents with only marginal effect on the receptor desensitization kinetics. In addition, BNC375 enhances long-term potentiation of electrically evoked synaptic responses in rat hippocampal slices and in vivo. Systemic administration of BNC375 reverses scopolamine-induced cognitive deficits in rat novel object recognition and rhesus monkey object retrieval detour (ORD) task over a wide range of exposures, showing no evidence of an inverted U-shaped dose-effect curve. The compound also improves performance in the ORD task in aged African green monkeys. Moreover, ex vivo 13C-NMR analysis indicates that BNC375 treatment can enhance neurotransmitter release in rat medial prefrontal cortex. These findings suggest that α7 nAChR PAMs have multiple advantages over orthosteric α7 nAChR agonists for the treatment of cognitive dysfunction associated with CNS diseases. SIGNIFICANCE STATEMENT: BNC375 is a novel and selective α7 nicotinic acetylcholine receptor (nAChR) positive allosteric modulator (PAM) that potentiates acetylcholine-evoked α7 currents in in vitro assays with little to no effect on the desensitization kinetics. In vivo, BNC375 demonstrated robust procognitive effects in multiple preclinical models across a wide exposure range. These results suggest that α7 nAChR PAMs have therapeutic potential in central nervous system diseases with cognitive impairments.

Fidelity to foraging sites after long migrations.

Patterns of animal movement associated with foraging lie at the heart of many ecological studies and often animals face decisions of staying in an environment they know versus relocating to new sites. The lack of knowledge of new foraging sites means there is risk associated with a decision to relocate (e.g. poor foraging) as well as a potential benefit (e.g. improved foraging). Using a unique long-term satellite tracking dataset for several sea turtle species, combined with capture-mark-recapture data extending over 50 years, we show how, across species, individuals generally maintain tight fidelity to specific foraging sites after extended (up to almost 10,000 km) migration to and from distant breeding sites as well as across many decades. Migrating individuals often travelled through suitable foraging areas en route to their 'home' site and so extended their journeys to maintain foraging site fidelity. We explore the likely mechanistic underpinnings of this trait, which is also seen in some migrating birds, and suggest that individuals will forgo areas of suitable forage encountered en route during migration when they have poor knowledge of the long-term suitability of those sites, making relocation to those sites risky.

Rapid and complete paraffin removal from human tissue sections delivers enhanced Raman spectroscopic and histopathological analysis.

Incomplete removal of paraffin and organic contaminants from tissues processed for diagnostic histology has been a profound barrier to the introduction of Raman spectroscopic techniques into clinical practice. We report a route to rapid and complete paraffin removal from a range of formalin-fixed paraffin embedded tissues using super mirror stainless steel slides. The method is equally effective on a range of human and animal tissues, performs equally well with archived and new samples and is compatible with standard pathology lab procedures. We describe a general enhancement of the Raman scatter and enhanced staining with antibodies used in immunohistochemistry for clinical diagnosis. We conclude that these novel slide substrates have the power to improve diagnosis through anatomical pathology by facilitating the simultaneous combination of improved, more sensitive immunohistochemical staining and simplified, more reliable Raman spectroscopic imaging, analysis and signal processing.

Effective hardness of interaction from thermodynamics and viscosity in dilute gases.

The hardness of the effective inverse power law (IPL) potential, which can be obtained from thermodynamics or collision integrals, can be used to demonstrate similarities between thermodynamic and transport properties. This link is investigated for systems of increasing complexity (i.e., the EXP, square-well, Lennard-Jones, and Stockmayer potentials; ab initio results for small molecules; and rigid linear chains of Lennard-Jones sites). These results show that while the two approaches do not yield precisely the same values of effective IPL exponent, their qualitative behavior is intriguingly similar, offering a new way of understanding the effective interactions between molecules, especially at high temperatures. In both approaches, the effective hardness is obtained from a double-logarithmic temperature derivative of an effective area.

An entropy scaling demarcation of gas- and liquid-like fluid behaviors.

In this work, we propose a generic and simple definition of a line separating gas-like and liquid-like fluid behaviors from the standpoint of shear viscosity. This definition is valid even for fluids such as the hard sphere and the inverse power law that exhibit a unique fluid phase. We argue that this line is defined by the location of the minimum of the macroscopically scaled viscosity when plotted as a function of the excess entropy, which differs from the popular Widom lines. For hard sphere, Lennard-Jones, and inverse-power-law fluids, such a line is located at an excess entropy approximately equal to -2/3 times Boltzmann's constant and corresponds to points in the thermodynamic phase diagram for which the kinetic contribution to viscosity is approximately half of the total viscosity. For flexible Lennard-Jones chains, the excess entropy at the minimum is a linear function of the chain length. This definition opens a straightforward route to classify the dynamical behavior of fluids from a single thermodynamic quantity obtainable from high-accuracy thermodynamic models.

Entropy Scaling of Viscosity - II: Predictive Scheme for Normal Alkanes.

In this work, a residual entropy value 6/10 of the way between the critical point and a value of -2/3 of Boltzmann's constant is shown to collapse the scaled viscosity for the family of normal alkanes. Based on this approach, a nearly universal correlation is proposed that can reproduce 95% of the experimental data for normal alkanes within ±18% (without removal of clearly erroneous data). This universal correlation has no new fluid-specific empirical parameters and is based on experimentally accessible values. This collapse is shown to be valid to a residual entropy half way between the critical point and the triple point, beyond which the macroscopically-scaled viscosity has a super-exponential dependence on residual entropy, terminating at the triple point. A key outcome of this study is a better understanding of entropy scaling for fluids with intramolecular degrees of freedom. A study of the transport and thermodynamic properties at the triple point rounds out the analysis.

Entropy Scaling of Viscosity - I: A Case Study of Propane.

In this work, a broadly-applicable and simple approach for building high accuracy viscosity correlations is demonstrated for propane. The approach is based on the combination of a number of recent insights related to the use of residual entropy scaling, especially a new way of scaling the viscosity for consistency with the dilute-gas limit. With three adjustable parameters in the dense phase, the primary viscosity data for propane are predicted with a mean absolute relative deviation of 1.38%, and 95% of the primary data are predicted within a relative error band of less than 5%. The dimensionality of the dense-phase contribution is reduced from the conventional two dimensional approach (temperature and density) to a one-dimensional correlation with residual entropy as the independent variable. The simplicity of the model formulation ensures smooth extrapolation behavior (barring errors in the equation of state itself). The approach proposed here should be applicable to a wide range of chemical species. The supporting information includes the relevant data in tabular form and a Python implementation of the model.

The Zero-Density Limit of the Residual Entropy Scaling of Transport Properties.

The modified residual entropy scaling approach has been shown to be a successful means of scaling dense phase transport properties. In this work, we investigate the dilute-gas limit of this scaling. This limit is considered for model potentials and highly accurate results from calculations with ab initio pair potentials for small molecules. These results demonstrate that with this approach, the scaled transport properties of noble gases can be collapsed without any empirical parameters to nearly their mutual uncertainties and that the scaled transport properties of polyatomic molecules are qualitatively similar, and for sufficiently high temperatures they agree with "universal" values proposed by Rosenfeld in 1999. There are significant quantitative differences between the model potentials and real fluids in these scaled coordinates, but this study provides a thorough coverage of model fluids and simple real fluids, providing the basis for further study. In the supporting information we provide the collected calculations with ab initio pair potentials from the literature, as well as code in the Python language implementing all aspects of our analysis.

The hunt for nonflammable refrigerant blends to replace R-134a.

We investigated refrigerant blends as possible low GWP (global warming potential) alternatives for R-134a in an air-conditioning application. We carried out an extensive screening of the binary, ternary, and four-component blends possible among a list of 13 pure refrigerants comprising four hydrofluoroolefins (HFOs), eight hydrofluorocarbons (HFCs), and carbon dioxide. The screening was based on a simplified cycle model, but with the inclusion of pressure drops in the evaporator and condenser. The metrics for the evaluation were nonflammability, low GWP, high COP (coefficient of performance), and a volumetric capacity similar to the R-134a baseline system. While no mixture was ideal in all regards, we identified 16 binary and ternary blends that were nonflammable (based on a new estimation method) and with COP and capacity similar to the R-134a baseline; the tradeoff, however, was a reduction in GWP of, at most, 54% compared to R-134a. An additional seven blends that were estimated to be "marginally flammable" (ASHRAE Standard 34 classification of A2L) were identified with GWP reductions of as much as 99%. These 23 "best" blends were then simulated in a more detailed cycle model.

Landscape of transcription in human cells.

Eukaryotic cells make many types of primary and processed RNAs that are found either in specific subcellular compartments or throughout the cells. A complete catalogue of these RNAs is not yet available and their characteristic subcellular localizations are also poorly understood. Because RNA represents the direct output of the genetic information encoded by genomes and a significant proportion of a cell's regulatory capabilities are focused on its synthesis, processing, transport, modification and translation, the generation of such a catalogue is crucial for understanding genome function. Here we report evidence that three-quarters of the human genome is capable of being transcribed, as well as observations about the range and levels of expression, localization, processing fates, regulatory regions and modifications of almost all currently annotated and thousands of previously unannotated RNAs. These observations, taken together, prompt a redefinition of the concept of a gene.