The impact of JUUL market entry on cigarette sales: evidence from a major chain retailer in Canada.

BACKGROUND: Electronic nicotine delivery systems (ENDS), such as the JUUL system, are nicotine products for adults who currently smoke cigarettes but are looking for an alternative to combustible cigarettes. Sales of ENDS products were legislatively acknowledged and authorized federally in Canada with the Royal Assent of the Tobacco and Vaping Products Act in 2018. METHODS: With the unique dataset from a major chain retailer in Canada, we evaluated the impacts of JUUL market entry on cigarette sales across Canada from January 2017 to August 2019 using two-way fixed effects panel regression models by leveraging on the entry time variation at the city level. We conducted various robustness checks and a permutation test to validate our results. RESULTS: Our estimates suggested that JUUL market entry was, on average, significantly correlated with a 1.65% per-month decrease in cigarette sales during the initial months, and with a potentially larger impact on urban areas. Our results were robust across various specifications and tests. These findings implied that JUUL and combustible cigarettes act as economic substitutes during the study time period in Canada. CONCLUSIONS: These results suggested that local availability of ENDS products, such as JUUL, has the potential to reduce local cigarette consumption.

Drought at a coastal wetland affects refuelling and migration strategies of shorebirds.

Droughts can affect invertebrate communities in wetlands, which can have bottom-up effects on the condition and survival of top predators. Shorebirds, key predators at coastal wetlands, have experienced widespread population declines and could be negatively affected by droughts. We explored, in detail, the effects of drought on multiple aspects of shorebird stopover and migration ecology by contrasting a year with average wet/dry conditions (2016) with a year with moderate drought (2017) at a major subarctic stopover site on southbound migration. We also examined the effects of drought on shorebird body mass during stopover across 14 years (historical: 1974-1982 and present-day: 2014-2018). For the detailed comparison of two years, in the year with moderate drought we documented lower invertebrate abundance at some sites, higher prey family richness in shorebird faecal samples, lower shorebird refuelling rates, shorter stopover durations for juveniles, and, for most species, a higher probability of making a subsequent stopover in North America after departing the subarctic, compared to the year with average wet/dry conditions. In the 14-year dataset, shorebird body mass tended to be lower in drier years. We show that even short-term, moderate drought conditions can negatively affect shorebird refuelling performance at coastal wetlands, which may carry-over to affect subsequent stopover decisions. Given shorebird population declines and predicted changes in the severity and duration of droughts with climate change, researchers should prioritize a better understanding of how droughts affect shorebird refuelling performance and survival.

Minimum Information and Quality Standards for Conducting, Reporting, and Organizing In Vitro Research.

Insufficient description of experimental practices can contribute to difficulties in reproducing research findings. In response to this, "minimum information" guidelines have been developed for different disciplines. These standards help ensure that the complete experiment is described, including both experimental protocols and data processing methods, allowing a critical evaluation of the whole process and the potential recreation of the work. Selected examples of minimum information checklists with relevance for in vitro research are presented here and are collected by and registered at the MIBBI/FAIRsharing Information Resource portal.In addition, to support integrative research and to allow for comparisons and data sharing across studies, ontologies and vocabularies need to be defined and integrated across areas of in vitro research. As examples, this chapter addresses ontologies for cells and bioassays and discusses their importance for in vitro studies.Finally, specific quality requirements for important in vitro research tools (like chemical probes, antibodies, and cell lines) are suggested, and remaining issues are discussed.

Baseline corticosterone does not reflect iridescent plumage traits in female tree swallows.

The production of high quality secondary sexual traits can be constrained by trade-offs in the allocation of energy and nutrients with other metabolic activities, and is mediated by physiological processes. In birds, the factors influencing male plumage quality have been well studied; however, factors affecting female plumage quality are poorly understood. Furthermore, it remains uncertain which physiological traits mediate the relationship between body condition and ornaments. In this three-year study of after-second-year female tree swallows (Tachycineta bicolor), we investigated (1) the relationship between baseline corticosterone near the end of the brood-rearing period (CORTBR) and feather colour characteristics (hue, saturation, brightness) the following year, and (2) the relationship between baseline corticosterone measured during incubation (CORTI) and brood rearing (CORTBR), and feather colour in the same year. To control for reproductive effort, we included reproductive parameters as covariates in all analyses. In this first study between CORT and the plumage colour characteristics of a species bearing iridescent feathers, we did not find any relationship between CORTBR and the colour of subsequently-produced feathers, nor did we find any relationship between CORT and the colour of feathers displayed during that breeding season. If CORT levels at the end of breeding carry over to influence the immediately subsequent moult period as we expect, our results generally indicate that structural plumage quality may not be as sensitive to circulating CORT levels compared to carotenoid-based colouration. Future studies, particularly those employing experimental manipulations of CORT during moult in species with iridescent traits, are necessary to fully determine the role glucocorticoids play in mediating the quality of secondary sexual characteristics.

An automated segmentation approach to calibrating infantile nystagmus waveforms.

Infantile nystagmus (IN) describes a regular, repetitive movement of the eyes. A characteristic feature of each cycle of the IN eye movement waveform is a period in which the eyes are moving at minimal velocity. This so-called "foveation" period has long been considered the basis for the best vision in individuals with IN. In recent years, the technology for measuring eye movements has improved considerably, but there remains the challenge of calibrating the direction of gaze in tracking systems when the eyes are continuously moving. Identifying portions of the nystagmus waveform suitable for calibration typically involves time-consuming manual selection of the foveation periods from the eye trace. Without an accurate calibration, the exact parameters of the waveform cannot be determined. In this study, we present an automated method for segmenting IN waveforms with the purpose of determining the foveation positions to be used for calibration of an eye tracker. On average, the "point of regard" was found to be within 0.21° of that determined by hand-marking by an expert observer. This method enables rapid clinical quantification of waveforms and the possibility of gaze-contingent research paradigms being performed with this patient group.

ABBV-105, a selective and irreversible inhibitor of Bruton's tyrosine kinase, is efficacious in multiple preclinical models of inflammation.

OBJECTIVES: Bruton's tyrosine kinase (BTK) is a non-receptor tyrosine kinase required for intracellular signaling downstream of multiple immunoreceptors. We evaluated ABBV-105, a covalent BTK inhibitor, using in vitro and in vivo assays to determine potency, selectivity, and efficacy to validate the therapeutic potential of ABBV-105 in inflammatory disease. METHODS: ABBV-105 potency and selectivity were evaluated in enzymatic and cellular assays. The impact of ABBV-105 on B cell function in vivo was assessed using mechanistic models of antibody production. Efficacy of ABBV-105 in chronic inflammatory disease was evaluated in animal models of arthritis and lupus. Measurement of BTK occupancy was employed as a target engagement biomarker. RESULTS: ABBV-105 irreversibly inhibits BTK, demonstrating superior kinome selectivity and is potent in B cell receptor, Fc receptor, and TLR-9-dependent cellular assays. Oral administration resulted in rapid clearance in plasma, but maintenance of BTK splenic occupancy. ABBV-105 inhibited antibody responses to thymus-independent and thymus-dependent antigens, paw swelling and bone destruction in rat collagen induced arthritis, and reduced disease in an IFNα-accelerated lupus nephritis model. BTK occupancy in disease models correlated with in vivo efficacy. CONCLUSION: ABBV-105, a selective BTK inhibitor, demonstrates compelling efficacy in pre-clinical mechanistic models of antibody production and in models of rheumatoid arthritis and lupus.

An evaluation of feather corticosterone as a biomarker of fitness and an ecologically relevant stressor during breeding in the wild.

Feather corticosterone (CORT) levels are increasingly employed as biomarkers of environmental stress. However, it is unclear if feather CORT levels reflect stress and/or workload in the wild. We investigated whether feather CORT represents a biomarker of environmental stress and reproductive effort in tree swallows (Tachycineta bicolor). Specifically, we examined whether individual state and investment during reproduction could predict feather CORT levels in subsequently moulted feathers and whether those levels could predict future survival and reproductive success. Through a manipulation of flight cost during breeding, we also investigated whether an increase in stress level would be reflected in subsequently grown feathers, and whether those levels could predict future success. We found that CORT levels of feathers grown during moult did not (1) reflect past breeding experience (n = 29), (2) predict reproductive output (n = 18), or (3) respond to a manipulation of flight effort during reproduction (10 experimental, 14 control females). While higher feather CORT levels predicted higher return rate (a proxy for survival), they did so only in the manipulated group (n = 36), and this relationship was opposite to expected. Overall, our results add to the mixed literature reporting that feather CORT levels can be positively, negatively, or not related to proxies of within-season and longer-term fitness (i.e., carryover effects). In addition, our results indicate that CORT levels or disturbances experienced during one time (e.g., breeding) may not carry over to subsequent stages (e.g., moult). We, therefore, petition for directed research investigating whether feather CORT represents exposure to chronic stress in feathers grown during moult.

Tricyclic covalent inhibitors selectively target Jak3 through an active site thiol.

The action of Janus kinases (JAKs) is required for multiple cytokine signaling pathways, and as such, JAK inhibitors hold promise for treatment of autoimmune disorders, including rheumatoid arthritis, inflammatory bowel disease, and psoriasis. However, due to high similarity in the active sites of the four members (Jak1, Jak2, Jak3, and Tyk2), developing selective inhibitors within this family is challenging. We have designed and characterized substituted, tricyclic Jak3 inhibitors that selectively avoid inhibition of the other JAKs. This is accomplished through a covalent interaction between an inhibitor containing a terminal electrophile and an active site cysteine (Cys-909). We found that these ATP competitive compounds are irreversible inhibitors of Jak3 enzyme activity in vitro. They possess high selectivity against other kinases and can potently (IC50 < 100 nm) inhibit Jak3 activity in cell-based assays. These results suggest irreversible inhibitors of this class may be useful selective agents, both as tools to probe Jak3 biology and potentially as therapies for autoimmune diseases.

Sphingosine-1-Phosphate (S1P) Lyase Inhibition Causes Increased Cardiac S1P Levels and Bradycardia in Rats.

Inhibition of the sphingosine-1-phosphate (S1P)-catabolizing enzyme S1P lyase (S1PL) elevates the native ligand of S1P receptors and provides an alternative mechanism for immune suppression to synthetic S1P receptor agonists. S1PL inhibition is reported to preferentially elevate S1P in lymphoid organs. Tissue selectivity could potentially differentiate S1PL inhibitors from S1P receptor agonists, the use of which also results in bradycardia, atrioventricular block, and hypertension. But it is unknown if S1PL inhibition would also modulate cardiac S1P levels or cardiovascular function. The S1PL inhibitor 6-[(2R)-4-(4-benzyl-7-chlorophthalazin-1-yl)-2-methylpiperazin-1-yl]pyridine-3-carbonitrile was used to determine the relationship in rats between drug concentration, S1P levels in select tissues, and circulating lymphocytes. Repeated oral doses of the S1PL inhibitor fully depleted circulating lymphocytes after 3 to 4 days of treatment in rats. Full lymphopenia corresponded to increased levels of S1P of 100- to 1000-fold in lymph nodes, 3-fold in blood (but with no change in plasma), and 9-fold in cardiac tissue. Repeated oral dosing of the S1PL inhibitor in telemeterized, conscious rats resulted in significant bradycardia within 48 hours of drug treatment, comparable in magnitude to the bradycardia induced by 3 mg/kg fingolimod. These results suggest that S1PL inhibition modulates cardiac function and does not provide immune suppression with an improved cardiovascular safety profile over fingolimod in rats.

Creation of a S1P Lyase bacterial surrogate for structure-based drug design.

S1P Lyase (SPL) has been described as a drug target in the treatment of autoimmune diseases. It plays an important role in maintaining intracellular levels of S1P thereby affecting T cell egress from lymphoid tissues. Several groups have already published approaches to inhibit S1P Lyase with small molecules, which in turn increase endogenous S1P concentrations resulting in immunosuppression. The use of structural biology has previously aided SPL inhibitor design. Novel construct design is at times necessary to provide a reagent for protein crystallography. Here we present a chimeric bacterial protein scaffold used for protein X-ray structures in the presence of early small molecule inhibitors. Mutations were introduced to the bacterial SPL from Symbiobacterium thermophilum which mimic the human enzyme. As a result, two mutant StSPL crystal structures resolved to 2.8Å and 2.2Å resolutions were solved and provide initial structural hypotheses for an isoxazole chemical series, whose optimization is discussed in the accompanying paper.

Hit-to-lead evaluation of a novel class of sphingosine 1-phosphate lyase inhibitors.

Inhibition of sphingosine-1-phosphate lyase has recently been proposed as a potential treatment option for inflammatory disorders such as multiple sclerosis, rheumatoid arthritis, and inflammatory bowel disease. In this report we describe our hit-to-lead evaluation of the isoxazolecarboxamide 6, a high-throughput screening hit (in vitro IC50=1.0 µM, cell IC50=1.8 µM), as a novel S1P lyase inhibitor. We were able to establish basic structure-activity relationships around 6 and succeeded in obtaining X-ray structural information which enabled structure-based design. With the discovery of 28, enzyme activity was quickly improved to IC50=120 nM and cell potency to IC50=230 nM. The main liability in the established isoxazolecarboxamide hit series was determined to be metabolic stability. In particular we identified that future lead-optimization efforts to overcome this problem should focus on blocking the N-dealkylation on the secondary amine.

Syk Inhibition Induces Platelet Dependent Peri-islet Hemorrhage in the Rat Pancreas.

Spleen tyrosine kinase (Syk) is a nonreceptor tyrosine kinase that is an important signaling enzyme downstream of immunoreceptors containing an intracellular immunoreceptor tyrosine activating motif (ITAM). These receptors encompass a wide variety of biological functions involved in autoimmune disease pathogenesis. There has been considerable interest in the development of inhibitors of the Syk pathway for the treatment of rheumatoid arthritis and systemic lupus erythematosus. We report that Syk inhibition mechanistically caused peri-islet hemorrhages and fibrin deposition in the rat pancreas and that this finding is due to a homeostatic functional defect in platelets. In more limited studies, similar lesions could not be induced in mice, dogs, and cynomolgus monkeys at similar or higher plasma drug concentrations. Irradiation-induced thrombocytopenia caused a phenotypically similar peri-islet pancreas lesion and the formation of this lesion could be prevented by platelet transfusion. In addition, Syk inhibitor-induced lesions were prevented by the coadministration of prednisone. A relatively greater sensitivity of rat platelets to Syk inhibition was supported by functional analyses demonstrating rat-specific differences in response to convulxin, a glycoprotein VI agonist that signals through Syk. These data demonstrate that the Syk pathway is critical in platelet-endothelial cell homeostasis in the peri-islet pancreatic microvasculature in rats.

Human optokinetic nystagmus and spatial frequency.

Optokinetic nystagmus (OKN) is a fundamental oculomotor response to retinal slip generated during natural movement through the environment. The timing and amplitude of the compensatory slow phases (SPs) alternating with saccadic quick phases (QPs) are remarkably variable, producing a characteristic irregular sawtooth waveform. We have previously found three stochastic processes that underlie OKN: the processes that determine QP and SP amplitude and the update dynamics of SP velocity. SP and QP parameters are interrelated and dependent on SP velocity such that changes in stimulus speed can have a seemingly complex effect on the nystagmus waveform. In this study we investigated the effect of stimulus spatial frequency on the stochastic processes of OKN. We found that increasing the spatial frequency of suprathreshold stimuli resulted in a significant increase in SP velocity with a corresponding reduction in retinal slip. However, retinal slip rarely reached values close to 0, indicating that the OKN system does not or cannot always minimize retinal slip. We deduce that OKN gain must be less than unity if extraretinal gain is lower than unity (as empirically observed), and that the difference between retinal and extraretinal gain determines the Markov properties of SP velocity. As retinal gain is reduced with stimuli of lower spatial frequency, the difference between retinal and extraretinal gain increases and the Markov properties of the system can be observed.

The distribution of quick phase interval durations in human optokinetic nystagmus.

There is an interesting dichotomy between models that predict the quick phase interval durations (QPIDs) of human optokinetic nystagmus (OKN). Accumulator models describe a stochastic signal in a neural network that triggers a response once the signal reaches a fixed threshold value. However, it is also possible that quick phases are triggered after eye position reaches a variable amplitude threshold. In this study, we fitted a range of probability density functions previously predicted by stochastic models of OKN (including those of the reciprocal truncated Normal, inverse Gaussian, gamma, lognormal and the mixture of two reciprocal truncated Normal distributions) to individual QPID histograms. We compared the goodness of fit between these models, and a model where the distribution of QPIDs is determined by the ratio of two correlated and truncated Normal random variables. The ratio distribution gave the best fit to the data, and we propose this is due to the approximately linear trajectory of slow phases (SPs) and that QPIDs are given by the ratio of a variable SP amplitude threshold and variable SP velocity.

Analysis of Fibroblast Growth Factor 14 (FGF14) structural variants reveals the genetic basis of the early onset nystagmus locus NYS4 and variable ataxia.

Nystagmus (involuntary, rhythmical eye movements) can arise due to sensory eye defects, in association with neurological disorders or as an isolated condition. We identified a family with early onset nystagmus and additional neurological features carrying a partial duplication of FGF14, a gene associated with spinocerebellar ataxia type 27 (SCA27) and episodic ataxia. Detailed eye movement analysis revealed oculomotor anomalies strikingly similar to those reported in a previously described four-generation family with early onset nystagmus and linkage to a region on chromosome 13q31.3-q33.1 (NYS4). Since FGF14 lies within NYS4, we revisited the original pedigree using whole genome sequencing, identifying a 161 kb heterozygous deletion disrupting FGF14 and ITGBL1 in the affected individuals, suggesting an FGF14-related condition. Therefore, our study reveals the genetic variant underlying NYS4, expands the spectrum of pathogenic FGF14 variants, and highlights the importance of screening FGF14 in apparently isolated early onset nystagmus.

A colonial-nesting seabird shows limited heart rate responses to natural variation in threats of polar bears.

Several predator-prey systems are in flux as an indirect result of climate change. In the Arctic, earlier sea-ice loss is driving polar bears (Ursus maritimus) onto land when many colonial nesting seabirds are breeding. The result is a higher threat of nest predation for birds with potential limited ability to respond. We quantified heart rate change in a large common eider (Somateria mollissima) breeding colony in the Canadian Arctic to explore their adaptive capacity to keep pace with the increasing risk of egg predation by polar bears. Eiders displayed on average higher heart rates from baseline when polar bears were within their field of view. Moreover, eiders were insensitive to variation in the distance bears were to their nests, but exhibited mild bradycardia (lowered heart rate) the longer the eider was exposed to the bear given the hen's visibility. Results indicate that a limited ability to assess the risks posed by polar bears may result in long-term fitness consequences for eiders from the increasing frequency in interactions with this predator.

Future Doctors From Underrepresented in Medicine Communities at the University of Utah.

Background and Objectives: Future Doctors (FD), a high school pathway program, was developed to address the lack of compositional diversity in the health professions at our health sciences campus. Methods: We obtained, analyzed, and compared data on FD student demographic and educational achievement at undergraduate and graduate programs at the University of Utah and graduate programs at other institutions to non-FD students. We followed students from high school to graduate school. Results: We analyzed data from 1,897 FD participants (2014-2019). FD participants were 71% women, with 50.3% identifying as students of color. Ninety-eight students matriculated in graduate school, with 75 (76%) remaining at University of Utah and 45 (46%) attending health sciences graduate programs. Conclusions: FD student cohorts are more diverse than those in the University of Utah undergraduate and graduate programs, contributing to the diversity of those programs. More research is needed to ensure that graduate school gains are evidenced in all underrepresented groups.

Human optokinetic nystagmus: a stochastic analysis.

Optokinetic nystagmus (OKN) is a fundamental gaze-stabilizing response found in almost all vertebrates, in which eye movements attempt to compensate for the optic flow caused by self-motion. It is an alternating sequence of slow compensatory eye movements made in the direction of stimulus motion and fast eye movements made predominantly in the opposite direction. The timing and amplitude of these slow phases (SPs) and quick phases (QPs) are remarkably variable, and the cause of this variability is poorly understood. In this study principal components analysis was performed on OKN data to illustrate that the variability in correlation matrices across individuals and recording sessions reflected changes in the noise in the system while the linear relationships between variables remained predominantly the same. Three components were found that could explain the variance in OKN data, and only variables from within a single cycle contributed highly to any given component. A linear stochastic model of OKN was developed based on these results that describes OKN as a triple first order Markov process, with three sources of noise affecting SP velocity, the QP trigger, and QP amplitude. This model was used to predict the degree of signal dependent noise in the system, the duration of the transient state of SP velocity, and an apparent undershoot bias to the QP target location.

A colonial-nesting seabird shows no heart-rate response to drone-based population surveys.

Aerial drones are increasingly being used as tools for ecological research and wildlife monitoring in hard-to-access study systems, such as in studies of colonial-nesting birds. Despite their many advantages over traditional survey methods, there remains concerns about possible disturbance effects that standard drone survey protocols may have on bird colonies. There is a particular gap in the study of their influence on physiological measures of stress. We measured heart rates of incubating female common eider ducks (Somateria mollissima) to determine whether our drone-based population survey affected them. To do so, we used heart-rate recorders placed in nests to quantify their heart rate in response to a quadcopter drone flying transects 30 m above the nesting colony. Eider heart rate did not change from baseline (measured in the absence of drone survey flights) by a drone flying at a fixed altitude and varying horizontal distances from the bird. Our findings suggest that carefully planned drone-based surveys of focal species have the potential to be carried out without causing physiological impacts among colonial-nesting eiders.

Thienopyrrole acetic acids as antagonists of the CRTH2 receptor.

The bioisosteric replacement of the indole core of CRTH2 antagonists using thienopyrroles was investigated, resulting in potent antagonists with good selectivity over DP1. Early ADME/PK assessment of this chemotype demonstrated bioavailability in mice.