Structural basis for selectivity and diversity in angiotensin II receptors.

The angiotensin II receptors AT1R and AT2R serve as key components of the renin-angiotensin-aldosterone system. AT1R has a central role in the regulation of blood pressure, but the function of AT2R is unclear and it has a variety of reported effects. To identify the mechanisms that underlie the differences in function and ligand selectivity between these receptors, here we report crystal structures of human AT2R bound to an AT2R-selective ligand and to an AT1R/AT2R dual ligand, capturing the receptor in an active-like conformation. Unexpectedly, helix VIII was found in a non-canonical position, stabilizing the active-like state, but at the same time preventing the recruitment of G proteins or ß-arrestins, in agreement with the lack of signalling responses in standard cellular assays. Structure-activity relationship, docking and mutagenesis studies revealed the crucial interactions for ligand binding and selectivity. Our results thus provide insights into the structural basis of the distinct functions of the angiotensin receptors, and may guide the design of new selective ligands.

Parabolic achromatic color matching functions: Dependence on incremental and decremental luminance.

Either the brightness or lightness of a disk surrounded by an annulus is characterized in the most general case by a parabolic function of the annulus luminance when plotted on a log-log scale. This relationship has been modeled with a theory of achromatic color computation based on edge integration and contrast gain control [J. Vis.10, 1 (2010)1534-736210.1167/10.14.40]. We tested predictions of this model in new psychophysical experiments. Our results support the theory and reveal a previously unobserved property of parabolic matching functions that depends on the disk contrast polarity. We interpret this property in terms of a neural edge integration model incorporating data from macaque monkey physiology that indicates different physiological gain factors for incremental and decremental stimuli.

Exploring the Determinants of Color Perception Using #Thedress and Its Variants: The Role of Spatio-Chromatic Context, Chromatic Illumination, and Material-Light Interaction.

The colors that people see depend not only on the surface properties of objects but also on how these properties interact with light as well as on how light reflected from objects interacts with an individual's visual system. Because individual visual systems vary, the same visual stimulus may elicit different perceptions from different individuals. #thedress phenomenon drove home this point: different individuals viewed the same image and reported it to be widely different colors: blue and black versus white and gold. This phenomenon inspired a collection of demonstrations presented at the Vision Sciences Society 2015 Meeting which showed how spatial and temporal manipulations of light spectra affect people's perceptions of material colors and illustrated the variability in individual color perception. The demonstrations also explored the effects of temporal alterations in metameric lights, including Maxwell's Spot, an entoptic phenomenon. Crucially, the demonstrations established that #thedress phenomenon occurs not only for images of the dress but also for the real dress under real light sources of different spectral composition and spatial configurations.

Brightness in human rod vision depends on slow neural adaptation to quantum statistics of light.

In human rod-mediated vision, threshold for small, brief flashes rises in proportion to the square root of adapting luminance at all but the lowest and highest adapting intensities. A classical signal detection theory from Rose (1942, 1948) and de Vries (1943) attributed this rise to the perceptual masking of weak flashes by Poisson fluctuations in photon absorptions from the adapting field. However, previous work by Brown and Rudd (1998) demonstrated that the square-root law also holds for suprathreshold brightness judgments, a finding that supports an alternative explanation of the square-root sensitivity changes as a consequence of physiological adaptation (i.e., neural gain control). Here, we employ a dichoptic matching technique to investigate the properties of this brightness gain control. We show that the brightness gain control: 1) affects the brightness of high-intensity suprathreshold flashes for which assumptions of the de Vries-Rose theory are strongly violated; 2) exhibits a long time course of 100-200 s; and 3) is subject to modulation by temporal contrast noise when the mean adapting luminance is held constant. These findings are consistent with the hypothesis that the square-root law results from a slow neural adaptation to statistical noise in the rod pool. We suggest that such adaptation may function to reduce the probability of spurious ganglion cell spiking activity due to photon fluctuation noise as the ambient illumination level is increased.

Acute vs. chronic stressors, multiple suicide attempts, and persistent suicide ideation in US soldiers.

This study examined recent-onset (i.e., acute) and persistent (i.e., chronic) life stressors among 54 acutely suicidal US Army Soldiers and examined their relationship to persistence of suicidal crises over time. Soldiers with a history of multiple suicide attempts reported the most severe suicide ideation (F(2,51) = 4.18, p = 0.021) and the greatest number of chronic stressors (F(2,51) = 5.11, p = 0.009). Chronic but not acute stressors were correlated with severity of suicide ideation (r = 0.24, p = 0.026). Participants reporting low-to-average levels of chronic stress resolved suicide ideation during the 6-month follow-up, but participants reporting high levels of chronic stress did not (Wald χ(1) = 4.57, p = 0.032). Soldiers who are multiple attempters report a greater number of chronic stressors. Chronic, but not acute-onset, stressors are associated with more severe and longer-lasting suicidal crises.

Loss of consciousness, depression, posttraumatic stress disorder, and suicide risk among deployed military personnel with mild traumatic brain injury.

OBJECTIVE: To identify clinical variables associated with suicidality in military personnel with mild traumatic brain injury (mTBI) while deployed to Iraq. SETTING: Outpatient TBI clinic on a US military base in Iraq. PARTICIPANTS: Military personnel (N = 158) referred to an outpatient TBI clinic for a standardized intake evaluation, 135 (85.4%) who had a diagnosis of mTBI and 23 (14.6%) who did not meet criteria for TBI. MAIN MEASURES: Suicidal Behaviors Questionnaire-Revised, Depression subscale of the Behavioral Health Measure-20, Posttraumatic Stress Disorder Checklist-Military Version, Insomnia Severity Index, self-report questionnaire, and clinical interview addressing TBI-related symptoms. RESULTS: Among patients with mTBI, increased suicidality was significantly associated with depression and the interaction of depression with posttraumatic stress disorder symptoms. Longer duration of loss of consciousness was associated with decreased likelihood for any suicidality. CONCLUSION: Assessment after TBI in a combat zone may assist providers in identifying those at risk for suicidality and making treatment recommendations for service members with mTBI.

Edge integration in achromatic color perception and the lightness-darkness asymmetry.

To maintain color constancy, the human visual system must distinguish surface reflectance-based variations in wavelength and luminance from variations due to illumination. Edge integration theory proposes that this is accomplished by spatially integrating steps in luminance and color contrast that likely result from reflectance changes. Thus, a neural representation of relative reflectance within the visual scene is constructed. An anchoring rule-the largest reflectance in the neural representation appears white-is then applied to map relative lightness onto an absolute lightness scale. A large body of data on human lightness judgments is here shown to be consistent with an edge integration model in which the visual system performs a weighted sum of steps in log luminance across space. Three hypotheses are proposed regarding how weights are applied to edges. First, weights decline with distance from the target surface whose lightness is being computed. Second, larger weights are given to edges whose dark sides point towards the target. Third, edge integration is carried out along a path leading from a common background field, or surround, to the target location. The theory accounts for simultaneous contrast; quantitative lightness judgments made with classical disk-annulus, Gilchrist dome, and Gelb displays; and perceptual filling-in lightness. A cortical theory of lightness in the ventral stream of visual cortex (areas V1 → V4) is proposed to instantiate the edge integration algorithm. The neural model is shown to be capable of unifying the quantitative laws of edge integration in lightness perception with the laws governing brightness, including Stevens' power law brightness model, and makes novel predictions about the quantitative laws governing induced darkness.

Discovery of MK-8768, a Potent and Selective mGluR2 Negative Allosteric Modulator.

Glutamate plays a key role in cognition and mood, and it has been shown that inhibiting ionotropic glutamate receptors disrupts cognition, while enhancing ionotropic receptor activity is pro-cognitive. One approach to elevating glutamatergic tone has been to antagonize presynaptic metabotropic glutamate receptor 2 (mGluR2). A desire for selectivity over the largely homologous mGluR3 motivated a strategy to achieve selectivity through the identification of mGluR2 negative allosteric modulators (NAMs). Extensive screening and optimization efforts led to the identification of a novel series of 4-arylquinoline-2-carboxamides. This series was optimized for mGluR2 NAM potency, clean off-target activity, and desirable physical properties, which resulted in the identification of improved C4 and C7 substituents. The initial lead compound from this series was Ames-positive in a single strain with metabolic activation, indicating that a reactive metabolite was likely responsible for the genetic toxicity. Metabolic profiling and Ames assessment across multiple analogs identified key structure-activity relationships associated with Ames positivity. Further optimization led to the Ames-negative mGluR2 negative allosteric modulator MK-8768.

MK-5172, a selective inhibitor of hepatitis C virus NS3/4a protease with broad activity across genotypes and resistant variants.

HCV NS3/4a protease inhibitors are proven therapeutic agents against chronic hepatitis C virus infection, with boceprevir and telaprevir having recently received regulatory approval as add-on therapy to pegylated interferon/ribavirin for patients harboring genotype 1 infections. Overcoming antiviral resistance, broad genotype coverage, and a convenient dosing regimen are important attributes for future agents to be used in combinations without interferon. In this communication, we report the preclinical profile of MK-5172, a novel P2-P4 quinoxaline macrocyclic NS3/4a protease inhibitor currently in clinical development. The compound demonstrates subnanomolar activity against a broad enzyme panel encompassing major hepatitis C virus (HCV) genotypes as well as variants resistant to earlier protease inhibitors. In replicon selections, MK-5172 exerted high selective pressure, which yielded few resistant colonies. In both rat and dog, MK-5172 demonstrates good plasma and liver exposures, with 24-h liver levels suggestive of once-daily dosing. When administered to HCV-infected chimpanzees harboring chronic gt1a or gt1b infections, MK-5172 suppressed viral load between 4 to 5 logs at a dose of 1 mg/kg of body weight twice daily (b.i.d.) for 7 days. Based on its preclinical profile, MK-5172 is anticipated to be broadly active against multiple HCV genotypes and clinically important resistance variants and highly suited for incorporation into newer all-oral regimens.

Development of potent macrocyclic inhibitors of genotype 3a HCV NS3/4A protease.

A series of macrocyclic compounds containing 2-substituted-quinoline moieties have been discovered and shown to exhibit excellent HCV NS3/4a genotype 3a and genotype 1b R155K mutant activity while maintaining the high rat liver exposure. Cyclization of the 2-substituted quinoline substituent led to a series of tricyclic P2 compounds which also display superb gt3a potency.

Development of macrocyclic inhibitors of HCV NS3/4A protease with cyclic constrained P2-P4 linkers.

A series of macrocyclic compounds containing a cyclic constraint in the P2-P4 linker region have been discovered and shown to exhibit excellent HCV NS3/4a genotype 3a and genotype 1b R155K, A156T, A156V, and D168V mutant activity while maintaining high rat liver exposure. The effect of the constraint is most dramatic against gt 1b A156 mutants where ~20-fold improvements in potency are achieved by introduction of a variety of ring systems into the P2-P4 linker.

Therapeutic drug monitoring of beta-lactam antibiotics in burns patients--a one-year prospective study.

BACKGROUND: Beta-lactams are first-line antibiotics for the management of superficial infections due to burn injury. There is sparse data available on therapeutic drug monitoring (TDM) in patients with burns in a ward setting. This study was conducted to evaluate the utility of a beta-lactam TDM program in a cohort of burn injury patients in a ward environment. METHODS: Steady-state blood samples were collected immediately before a scheduled dose. The therapeutic concentration targets assessed were (1) free antibiotic concentrations exceeding the minimum inhibitory concentration (MIC; fT > MIC) and (2) free concentrations ≥4× MIC of the known or suspected pathogen (fT > 4× MIC). The duration of therapy was also assessed. RESULTS: A total of 50 patients were included for TDM over a 12-month period. The mean (±SD) age was 49 ± 16 years. The mean percent total body surface area burn was 17 ± 13%. The mean serum creatinine concentration was 86 ± 20 µmole/L. Sixty percent of the patients did not achieve fT > MIC, and only 18% achieved the higher target of fT > 4× MIC. Although all the patients achieved a positive clinical outcome, the duration of antibiotic treatment was shorter in patients who achieved fT > MIC compared with those who did not (4.2 ± 1.1 versus 5.3 ± 2.3 days; P = 0.03). CONCLUSIONS: We found TDM to be a reliable intervention for burn injury patients in a ward environment. This study supports pharmacokinetic data that burns patients may be at risk of subtherapeutic dosing, which may prolong the duration of antibiotic therapy.

Circadian Rhythmicity of Vital Signs at Intensive Care Unit Discharge and Outcome of Traumatic Brain Injury.

BACKGROUND: Physiological functions with circadian rhythmicity are often disrupted during illness. OBJECTIVE: To assess the utility of circadian rhythmicity of vital signs in predicting outcome of traumatic brain injury (TBI). METHODS: A retrospective single-center cohort study of adult intensive care unit (ICU) patients with largely isolated TBI to explore the relationship between the circadian rhythmicity of vital signs during the last 24 hours before ICU discharge and clinical markers of TBI severity and score on the Glasgow Outcome Scale 6 months after injury (GOS-6). RESULTS: The 130 study participants had a median age of 39.0 years (IQR, 23.0-59.0 years), a median Glasgow Coma Scale score at the scene of 8.0 (IQR, 3.0-13.0), and a median Rotterdam score on computed tomography of the head of 3 (IQR, 3-3), with 105 patients (80.8%) surviving to hospital discharge. Rhythmicity was present for heart rate (30.8% of patients), systolic blood pressure (26.2%), diastolic blood pressure (20.0%), and body temperature (26.9%). Independent predictors of a dichotomized GOS-6 ≥4 were the Rotterdam score (odds ratio [OR], 0.38 [95% CI, 0.18-0.81]; P = .01), Glasgow Coma Scale score at the scene (OR, 1.22 [95% CI, 1.05-1.41]; P = .008), age (OR, 0.95 [95% CI, 0.92-0.98]; P = .003), oxygen saturation <90% in the first 24 hours (OR, 0.19 [95% CI, 0.05-0.73]; P = .02), serum sodium level <130 mmol/L (OR, 0.20 [95% CI, 0.05-0.70]; P = .01), and active intracranial pressure management (OR, 0.16 [95% CI, 0.04-0.62]; P = .008), but not rhythmicity of any vital sign. CONCLUSION: Circadian rhythmicity of vital signs at ICU discharge is not predictive of GOS-6 in patients with TBI.

The PET tracer [11C]MK-6884 quantifies M4 muscarinic receptor in rhesus monkeys and patients with Alzheimer's disease.

Positron emission tomography (PET) ligands play an important role in the development of therapeutics by serving as target engagement or pharmacodynamic biomarkers. Here, we describe the discovery and translation of the PET tracer [11C]MK-6884 from rhesus monkeys to patients with Alzheimer's disease (AD). [3H]MK-6884/[11C]MK-6884 binds with high binding affinity and good selectivity to an allosteric site on M4 muscarinic cholinergic receptors (M4Rs) in vitro and shows a regional distribution in the brain consistent with M4R localization in vivo. The tracer demonstrates target engagement of positive allosteric modulators of the M4R (M4 PAMs) through competitive binding interactions. [11C]MK-6884 binding is enhanced in vitro by the orthosteric M4R agonist carbachol and indirectly in vivo by the acetylcholinesterase inhibitor donepezil in rhesus monkeys and healthy volunteers, consistent with its pharmacology as a highly cooperative M4 PAM. PET imaging of [11C]MK-6884 in patients with AD identified substantial regional differences quantified as nondisplaceable binding potential (BPND) of [11C]MK-6884. These results suggest that [11C]MK-6884 is a useful target engagement biomarker for M4 PAMs but may also act as a sensitive probe of neuropathological changes in the brains of patients with AD.

Duration Comparisons for Vision and Touch Are Dependent on Presentation Order and Temporal Context.

Integrating visual and tactile information in the temporal domain is critical for active perception. To accomplish this, coordinated timing is required. Here, we study perceived duration within and across these two modalities. Specifically, we examined how duration comparisons within and across vision and touch were influenced by temporal context and presentation order using a two-interval forced choice task. We asked participants to compare the duration of two temporal intervals defined by tactile or visual events. Two constant standard durations (700 ms and 1,000 ms in 'shorter' sessions; 1,000 ms and 1,500 ms in 'longer' sessions) were compared to variable comparison durations in different sessions. In crossmodal trials, standard and comparison durations were presented in different modalities, whereas in the intramodal trials, the two durations were presented in the same modality. The standard duration was either presented first () or followed the comparison duration (). In both crossmodal and intramodal conditions, we found that the longer standard duration was overestimated in trials and underestimated in trials whereas the estimation of shorter standard duration was unbiased. Importantly, the estimation of 1,000ms was biased when it was the longer standard duration within the shorter sessions but not when it was the shorter standard duration within the longer sessions, indicating an effect of temporal context. The effects of presentation order can be explained by a central tendency effect applied in different ways to different presentation orders. Both crossmodal and intramodal conditions showed better discrimination performance for trials than trials, supporting the Type B effect for both crossmodal and intramodal duration comparison. Moreover, these results were not dependent on whether the standard duration was defined using tactile or visual stimuli. Overall, our results indicate that duration comparison between vision and touch is dependent on presentation order and temporal context, but not modality.

Structures of active-state orexin receptor 2 rationalize peptide and small-molecule agonist recognition and receptor activation.

Narcolepsy type 1 (NT1) is a chronic neurological disorder that impairs the brain's ability to control sleep-wake cycles. Current therapies are limited to the management of symptoms with modest effectiveness and substantial adverse effects. Agonists of the orexin receptor 2 (OX2R) have shown promise as novel therapeutics that directly target the pathophysiology of the disease. However, identification of drug-like OX2R agonists has proven difficult. Here we report cryo-electron microscopy structures of active-state OX2R bound to an endogenous peptide agonist and a small-molecule agonist. The extended carboxy-terminal segment of the peptide reaches into the core of OX2R to stabilize an active conformation, while the small-molecule agonist binds deep inside the orthosteric pocket, making similar key interactions. Comparison with antagonist-bound OX2R suggests a molecular mechanism that rationalizes both receptor activation and inhibition. Our results enable structure-based discovery of therapeutic orexin agonists for the treatment of NT1 and other hypersomnia disorders.

MK-7009, a potent and selective inhibitor of hepatitis C virus NS3/4A protease.

The administration of hepatitis C virus (HCV) NS3/4A protease inhibitors to patients with chronic HCV infections has demonstrated that they have dramatic antiviral effects and that compounds acting via this mechanism are likely to form a key component of future anti-HCV therapy. We report here on the preclinical profile of MK-7009, an inhibitor of genotype 1a and 1b proteases at subnanomolar concentrations with modestly shifted potency against genotype 2a and 2b proteases at low nanomolar concentrations. Potent activity was also observed in a cell-based HCV replicon assay in the presence of added human serum (50%). In multiple species evaluated in preclinical studies, the MK-7009 concentrations in the liver were maintained at a significant multiple of the cell-based replicon 50% effective concentration over 12 to 24 h following the administration of moderate oral doses (5 to 10 mg per kg of body weight). MK-7009 also had excellent selectivity against both a range of human proteases and a broad panel of pharmacologically relevant ion channels, receptors, and enzymes. On the basis of this favorable profile, MK-7009 was selected for clinical development and is currently being evaluated in controlled clinical trials with both healthy volunteers and HCV-infected patients.

How attention and contrast gain control interact to regulate lightness contrast and assimilation: a computational neural model.

Recent theories of lightness perception assume that lightness (perceived reflectance) is computed by a process that contrasts the target's luminance with that of one or more regions in its spatial surround. A challenge for any such theory is the phenomenon of lightness assimilation, which occurs when increasing the luminance of a surround region increases the target lightness: the opposite of contrast. Here contrast and assimilation are studied quantitatively in lightness matching experiments utilizing concentric disk-and-ring displays. Whether contrast or assimilation is seen depends on a number of factors including: the luminance relations of the target, surround, and background; surround size; and matching instructions. When assimilation occurs, it is always part of a larger pattern in which assimilation and contrast both occur over different ranges of surround luminance. These findings are quantitatively modeled by a theory that assumes lightness is computed from a weighted sum of responses of edge detector neurons in visual cortex. The magnitude of the neural response to an edge is regulated by a combination of contrast gain control acting between neighboring edge detectors and a top-down attentional gain control that selectively weights the response to stimulus edges according to their task relevance.

Neurocomputational Lightness Model Explains the Appearance of Real Surfaces Viewed Under Gelb Illumination.

One of the primary functions of visual perception is to represent, estimate, and evaluate the properties of material surfaces in the visual environment. One such property is surface color, which can convey important information about ecologically relevant object characteristics such as the ripeness of fruit and the emotional reactions of humans in social interactions. This paper further develops and applies a neural model (Rudd, 2013, 2017) of how the human visual system represents the light/dark dimension of color-known as lightness-and computes the colors of achromatic material surfaces in real-world spatial contexts. Quantitative lightness judgments conducted with real surfaces viewed under Gelb (i.e., spotlight) illumination are analyzed and simulated using the model. According to the model, luminance ratios form the inputs to ON- and OFF-cells, which encode local luminance increments and decrements, respectively. The response properties of these cells are here characterized by physiologically motivated equations in which different parameters are assumed for the two cell types. Under non-saturating conditions, ON-cells respond in proportion to a compressive power law of the local incremental luminance in the image that causes them to respond, while OFF-cells respond linearly to local decremental luminance. ON- and OFF-cell responses to edges are log-transformed at a later stage of neural processing and then integrated across space to compute lightness via an edge integration process that can be viewed as a neurally elaborated version of Land's retinex model (Land & McCann, 1971). It follows from the model assumptions that the perceptual weights-interpreted as neural gain factors-that the model observer applies to steps in log luminance at edges in the edge integration process are determined by the product of a polarity-dependent factor 1-by which incremental steps in log luminance (i.e., edges) are weighted by the value <1.0 and decremental steps are weighted by 1.0-and a distance-dependent factor 2, whose edge weightings are estimated to fit perceptual data. The model accounts quantitatively (to within experimental error) for the following: lightness constancy failures observed when the illumination level on a simultaneous contrast display is changed (Zavagno, Daneyko, & Liu, 2018); the degree of dynamic range compression in the staircase-Gelb paradigm (Cataliotti & Gilchrist, 1995; Zavagno, Annan, & Caputo, 2004); partial releases from compression that occur when the staircase-Gelb papers are reordered (Zavagno, Annan, & Caputo, 2004); and the larger compression release that occurs when the display is surrounded by a white border (Gilchrist & Cataliotti, 1994).

The Pre-Hospital Initial Fluid Therapy Estimate in Early Nasty Burns (PHIFTEEN B, 15-B) Guideline applied to a retrospective cohort of Intensive Care Unit patients with major burns.

BACKGROUND: Appropriate fluid administration in severe burns is a cornerstone of early burns management. The American Burns Association's (ABA) recommendation is to administer 2 mL-4 mL × burnt Body Surface Area (BSA) × weight in the first 24 h with half administered in the first eight hours. Unfortunately, the calculations involved are complex and clinicians do not estimate the BSA or weight well, which can lead to errors in the amount of fluid administered. To simplify cognitive load to calculate the fluid resuscitation of early burns, the investigators derived the PHIFTEEN B (15-B) guideline. The 15-B guideline estimates the initial hourly fluid for adults ≥ 50 kg to be: 15 mL × BSA (to the nearest 10%) AIMS: To model and determine the accuracy of the 15-B calculated based on the characteristics of a retrospective cohort of patients admitted with ≥ 20% BSA to the Royal Brisbane and Women's Hospital (RBWH) Intensive Care Unit (ICU). METHODS: The 15-B formula was retrospectively calculated on the prehospital BSA estimate on patients admitted to the RBWH ICU. In addition, the 15-B guideline was modelled against a variety of weights and BSAs. The fluid volume was deemed to be clinically significant if it was greater than 250 mL/h outside the ABA's recommendations. RESULTS: The ICU cohort consisted of 107 patients (63.2% male, median age 37 years), with a median ICU estimated BSA of 40% and a median ICU weight estimation of 80 kg. In 43.9% of the cohort, the magnitude of the proportional difference between prehospital and ICU BSA estimate was greater than 25%. The 15-B formula accurately estimated the hourly fluid for all BSA (20%-100%) and weight combinations (50 kg-140 kg) in a BSA- weight matrix. When prehospital BSA estimate was utilized, 15-B guideline accurately estimated the fluid to be given within clinically significant limits for 97.2% of cases. CONCLUSIONS: The 15-B formula is a simple, easy to calculate guideline which approximates the early fluid estimates in severely burned patients despite inaccuracy in prehospital BSA estimates.