Maximizing insights from nonclinical safety studies in the context of rising costs and changing regulations.

The traditional paradigm of non-rodent safety assessment studies, primarily reliant on non-human primates (NHPs) and dogs, is undergoing a transformation. During the 2023 Safety Pharmacology Society Annual Meeting, scientists from leading nonclinical contract organizations discussed how traditional IND-enabling studies can benefit from employing underutilized alternative non-rodent models, such as the swine. Swine offer a cost-effective approach to drug development and share many anatomical and physiological similarities with humans. The inclusion of non-traditional species in safety assessments, coupled with advanced measurement techniques, aids in de-risking compounds early on and adapting projects to the evolving cost landscape.

Ototoxicity: a high risk to auditory function that needs to be monitored in drug development.

Hearing loss constitutes a major global health concern impacting approximately 1.5 billion people worldwide. Its incidence is undergoing a substantial surge with some projecting that by 2050, a quarter of the global population will experience varying degrees of hearing deficiency. Environmental factors such as aging, exposure to loud noise, and the intake of ototoxic medications are implicated in the onset of acquired hearing loss. Ototoxicity resulting in inner ear damage is a leading cause of acquired hearing loss worldwide. This could be minimized or avoided by early testing of hearing functions in the preclinical phase of drug development. While the assessment of ototoxicity is well defined for drug candidates in the hearing field - required for drugs that are administered by the otic route and expected to reach the middle or inner ear during clinical use - ototoxicity testing is not required for all other therapeutic areas. Unfortunately, this has resulted in more than 200 ototoxic marketed medications. The aim of this publication is to raise awareness of drug-induced ototoxicity and to formulate some recommendations based on available guidelines and own experience. Ototoxicity testing programs should be adapted to the type of therapy, its indication (targeting the ear or part of other medications classes being potentially ototoxic), and the number of assets to test. For multiple molecules and/or multiple doses, screening options are available: in vitro (otic cell assays), ex vivo (cochlear explant), and in vivo (in zebrafish). In assessing the ototoxicity of a candidate drug, it is good practice to compare its ototoxicity to that of a well-known control drug of a similar class. Screening assays provide a streamlined and rapid method to know whether a drug is generally safe for inner ear structures. Mammalian animal models provide a more detailed characterization of drug ototoxicity, with a possibility to localize and quantify the damage using functional, behavioral, and morphological read-outs. Complementary histological measures are routinely conducted notably to quantify hair cells loss with cochleogram. Ototoxicity studies can be performed in rodents (mice, rats), guinea pigs and large species. However, in undertaking, or at the very least attempting, all preclinical investigations within the same species, is crucial. This encompasses starting with pharmacokinetics and pharmacology efficacy studies and extending through to toxicity studies. In life read-outs include Auditory Brainstem Response (ABR) and Distortion Product OtoAcoustic Emissions (DPOAE) measurements that assess the activity and integrity of sensory cells and the auditory nerve, reflecting sensorineural hearing loss. Accurate, reproducible, and high throughput ABR measures are fundamental to the quality and success of these preclinical trials. As in humans, in vivo otoscopic evaluations are routinely carried out to observe the tympanic membrane and auditory canal. This is often done to detect signs of inflammation. The cochlea is a tonotopic structure. Hair cell responsiveness is position and frequency dependent, with hair cells located close to the cochlea apex transducing low frequencies and those at the base transducing high frequencies. The cochleogram aims to quantify hair cells all along the cochlea and consequently determine hair cell loss related to specific frequencies. This measure is then correlated with the ABR & DPOAE results. Ototoxicity assessments evaluate the impact of drug candidates on the auditory and vestibular systems, de-risk hearing loss and balance disorders, define a safe dose, and optimize therapeutic benefits. These types of studies can be initiated during early development of a therapeutic solution, with ABR and otoscopic evaluations. Depending on the mechanism of action of the compound, studies can include DPOAE and cochleogram. Later in the development, a GLP (Good Laboratory Practice) ototoxicity study may be required based on otic related route of administration, target, or known potential otic toxicity.

Climate change, risk perception, and protection motivation among high-altitude residents of the Mt. Everest region in Nepal.

Mountain ecosystems are considered vulnerable to early impacts of climate change. Whether and how local residents of these areas perceive these changes, however, remain under-studied questions. By conducting a household survey in the Khumbu region of Nepal, this study assessed local residents' experience-based perception of changes in climate trends and patterns, perceived risk, and attitudes towards climate issues. Multivariate cluster analysis based on residents' climate change beliefs revealed three segments: "Cautious," "Disengaged," and "Alarmed." A comparison of these segments along key psychosocial constructs of Protection Motivation Theory (PMT) revealed significant inter-segment differences in residents' perception of severity, vulnerability, response efficacy, self-efficacy, and response cost associated with engaging in mitigating behavior. Results shed light on how residents of high elevation areas that are considered to be exposed to early impacts of climate change perceive the risk and intend to respond. These findings could also assist stakeholders working in other similar mountain ecosystems in understanding vulnerability and in working towards climate readiness.

Musicians' Earplugs: Do They Affect Performance or Listeners' Perceptions?

BACKGROUND: Does wearing musicians' earplugs while performing affect the quality of the performance? Can listeners perceive a difference in sound when musicians are performing with or without earplugs? The risk of hearing loss is a concern for musicians, but some are reluctant to wear hearing protection due to factors such as an inability to hear their own instrument properly and the possibility of decreased sound quality for listeners. OBJECTIVE: The purpose of this study was to determine the effect of musicians' earplugs on instrumental pitch accuracy and the perception of tone quality, intonation, and dynamic contrast, as perceived by musicians and listeners. METHODS: Ten university studio faculty teachers were recorded performing single pitches and lyrical and technical passages, first without earplugs and then immediately after with earplugs. A sample of 8 studio faculty teachers and 88 undergraduate music education students completed a researcher-created music perception test of tone quality, intonation, and dynamic contrast. RESULTS: Objective analyses of the single pitch recordings made by faculty with and without earplugs indicate that pitch accuracy did not favor either condition consistently. Results from the perception test indicate that although both faculty and student listeners perceived some differences, the most frequent perception was that the audio pair was equal, and there was no clear difference between performing with and without earplugs in terms of tone quality, intonation, or dynamic contrast. CONCLUSION: These findings suggest that musicians should feel confident that wearing musicians' earplugs while performing does not adversely affect pitch accuracy or listeners' perceptions of their timbre and dynamic control.

Dataset of forest landowner survey to assess interest in supplying woody biomass in two Southeastern United States fuelsheds.

This article presents data from a recent mail survey of forest landowners regarding their land ownership characteristics and motivations, past and future management activities, and owner perceptions of bioenergy and its impact on forests. The data presented in this article are related to the research article entitled 'Opportunities and Attitudes of Private Forest Landowners in Supplying Woody Biomass for Renewable Energy' [1]. The survey was conducted in the Atlantic Coastal Plain of the United States, where two primary ports exporting wood pellets to Europe are located. Specifically, the data include responses on forest characteristics, forest management activities, knowledge and interest in woody biomass for energy production, and sociodemographic variables. Additionally, landowner decisions for supplying wood for traditional forest products and biomass for energy were modeled. More than 2900 forest landowners were contacted, with 707 owners providing completed surveys.

The northeast regional SPS meeting update: Safety pharmacology innovations and applications.

The Safety Pharmacology Society (SPS) held a Northeast (NE) regional meeting in Boston, MA on May 13, 2016 at the Vertex Pharmaceuticals Incorporated site. There were 103 attendees from the pharmaceutical industry, contract research organizations (CROs), academia, and global regulatory agencies. An assortment of scientific topics were presented by 7 speakers that included broad topics in the cardiovascular (organ on chip, statistical power and translation of rat cardiovascular telemetry data and dual inhibition of IKr and IKs on QT interval prolongation) and central nervous system (in vitro platform for neurotoxicity, an integrated risk assessment of suicidal ideation and behavior, and EEG advances in safety pharmacology) and a novel topic discussing preclinical challenges faced in the development of a novel gene therapy. A highlight of the meeting was an in-depth discussion on the fatty acid acyl hydrolase (FAAH) inhibitor BIA 10-2474 which involved a comprehensive overview of the biology and pharmacology of FAAH followed by a presentation from the Biotrial (Rennes, France) team that conducted the clinical trial. An additional poster session was held that included 13 fascinating posters on cutting edge safety pharmacology topics.

Creativity and the default network: A functional connectivity analysis of the creative brain at rest.

The present research used resting-state functional magnetic resonance imaging (fMRI) to examine whether the ability to generate creative ideas corresponds to differences in the intrinsic organization of functional networks in the brain. We examined the functional connectivity between regions commonly implicated in neuroimaging studies of divergent thinking, including the inferior prefrontal cortex and the core hubs of the default network. Participants were prescreened on a battery of divergent thinking tests and assigned to high- and low-creative groups based on task performance. Seed-based functional connectivity analysis revealed greater connectivity between the left inferior frontal gyrus (IFG) and the entire default mode network in the high-creative group. The right IFG also showed greater functional connectivity with bilateral inferior parietal cortex and the left dorsolateral prefrontal cortex in the high-creative group. The results suggest that the ability to generate creative ideas is characterized by increased functional connectivity between the inferior prefrontal cortex and the default network, pointing to a greater cooperation between brain regions associated with cognitive control and low-level imaginative processes.

Music to the inner ears: exploring individual differences in musical imagery.

In two studies, we explored the frequency and phenomenology of musical imagery. Study 1 used retrospective reports of musical imagery to assess the contribution of individual differences to imagery characteristics. Study 2 used an experience sampling design to assess the phenomenology of musical imagery over the course of one week in a sample of musicians and non-musicians. Both studies found episodes of musical imagery to be common and positive: people rarely wanted such experiences to end and often heard music that was personally meaningful. Several variables predicted musical imagery, including personality, musical preferences, and positive mood. Musicians tended to hear musical imagery more often, but they reported less frequent episodes of deliberately-generated imagery. Taken together, the present research provides new insights into individual differences in musical imagery, and it supports the emerging view that such experiences are common, positive, and more voluntary than previously recognized.

Modeling transient response of forests to climate change.

Our hypothesis is that a high diversity of dominant life forms in Tennessee forests conveys resilience to disturbance such as climate change. Because of uncertainty in climate change and their effects, three climate change scenarios for 2030 and 2080 from three General Circulation Models (GCMs) were used to simulate a range of potential climate conditions for the state. These climate changes derive from the Intergovernmental Panel on Climate Change (IPCC) "A1B" storyline that assumes rapid global economic growth. The precipitation and temperature projections from the three GCMs for 2030 and 2080 were related to changes in five ecological provinces using the monthly record of temperature and precipitation from 1980 to 1997 for each 1km cell across the state as aggregated into the provinces. Temperatures are projected to increase in all ecological provinces in all months for all three GCMs for both 2030 and 2080. Precipitation differences from the long-term average are more complex but less striking. The forest ecosystem model LINKAGES was used to simulate conditions for five ecological provinces from 1989 to 2300. Average output projects changes in tree diversity and species composition in all ecological provinces in Tennessee with the greatest changes in the Southern Mixed Forest province. Projected declines in total tree biomass are followed by biomass recovery as species replacement occurs in stands. The Southern Mixed Forest province results in less diversity in dominant trees as well as lower overall biomass than projections for the other four provinces. The biomass and composition changes projected in this study differ from forest dynamics expected without climate change. These results suggest that biomass recovery following climate change is linked to dominant tree diversity in the southeastern forest of the US. The generality of this observation warrants further investigation, for it relates to ways that forest management may influence climate change effects.

Scopolamine induced deficits in a battery of rat cognitive tests: comparisons of sensitivity and specificity.

Despite much research, the cognitive effects of scopolamine hydrobromide, a cholinergic antagonist, remain controversial. Scopolamine affects multiple systems each of which can impact behavior. One way to tease apart the effects of the drug is to determine the effects of low scopolamine doses on different abilities. The present experiments compared the effects of low doses of scopolamine on a single group of rats conducting a battery of behavioral tasks: Morris water maze, radial arm maze, delayed non-matching to position tasks, and fixed ratio 5 bar pressing. The behavioral battery ranged from tasks having little cognitive demand to those thought to be based more on attention and spatial-working memory. Control experiments using additional groups of rats assessing peripheral versus central effects were conducted with both liquid and dry reinforcement and with methyl scopolamine. Furthermore, the 5-choice serial reaction time test assessed scopolamine effects on attention. The data show a wide spectrum of central and peripheral cholinergic involvement. The central effects include attention and motor initiation, both of which impact and interact with the mnemonic function of acetylcholine. These results show that a limited disruption of the central cholinergic system can have profound effects on attention and/or psychomotor control before any measurable mnemonic disruption.

Closing the mind's eye: deactivation of visual cortex related to auditory task difficulty.

Blood oxygen-level-dependent signal decreases relative to baseline (deactivations) can occur with stimulation of an opposing sensory modality. Here, we show the importance of the difficulty of an auditory task on the deactivation of visual cortical areas. Participants performed an auditory temporal-order judgment task in conjunction with sparse-sampling functional MRI at both moderate and high levels of difficulty (adjusted for each individual's own threshold). With moderate difficulty, small deactivations were observed not only in parietal and cingulate cortex, but occipital cortex as well. When the same task was more difficult, deactivations increased significantly to include a greater extent of functionally defined visual cortex. Together, these results suggest that cross-modal deactivations occur in compensation for task difficulty, perhaps acting as an intrinsic filter for nonrelevant information.

Donepezil primarily attenuates scopolamine-induced deficits in psychomotor function, with moderate effects on simple conditioning and attention, and small effects on working memory and spatial mapping.

RATIONALE: Alzheimer's dementia (AD) patients have profound deficits in cognitive and social functions, mediated in part by a decline in cholinergic function. Acetylcholinesterase inhibitors (AChEI) are the most commonly prescribed treatment for the cognitive deficits in AD patients, but their therapeutic effects are small, and it is still not clear if they primarily affect attention, memory, or some other cognitive/behavioral functions. OBJECTIVES: The objective of the present experiments was to explore the effects of donepezil (Aricepttrade mark), an AChEI, on behavioral deficits related exclusively to cholinergic dysfunction. MATERIALS AND METHODS: The effects of donepezil were assessed in Sprague-Dawley rats with scopolamine-induced deficits in a battery of cognitive/behavioral tests. RESULTS: Scopolamine produced deficits in contextual and cued fear conditioning, the 5-choice serial reaction time test, delayed nonmatching to position, the radial arm maze, and the Morris water maze. Analyses of the pattern and size of the effects revealed that donepezil produced very large effects on scopolamine-induced deficits in psychomotor function (approximately 20-50% of the variance), moderate-sized effects on scopolamine-induced deficits in simple conditioning and attention (approximately 3-10% of the variance), but only small effects on scopolamine-induced deficits in higher cognitive functions of working memory and spatial mapping (approximately 1% of the variance). CONCLUSIONS: These results are consistent with the limited efficacy of donepezil on higher cognitive function in AD patients, and suggest that preclinical behavioral models could be used not only to determine if novel treatments have some therapeutic potential, but also to predict more precisely what the pattern and size of the effects might be.

Soluble Abeta and cognitive function in aged F-344 rats and Tg2576 mice.

Recent findings suggest that Alzheimer's dementia may be mediated by soluble beta amyloid (Abeta) more than the deposits of aggregated, insoluble Abeta, and vulnerability to cognitive deficits after scopolamine challenge may help identify AD even in patients that are still pre-symptomatic. The objectives of the present experiments were to determine if vulnerability to cognitive deficits after scopolamine challenge is related to levels of soluble Abeta, and if levels of soluble Abeta are more closely related to cognitive deficits than levels of insoluble Abeta, even in aged, transgenic mice, after they have developed very high levels of insoluble Abeta. Aged F-344 rats and young mice over-expressing the Swedish mutation in the human amyloid precursor protein (APPsw; Tg2576+) had elevated levels of soluble Abeta, and were more vulnerable to scopolamine challenge in the Morris water maze (MWM), relative to young rats and Tg2576- mice; but, among individual animals, higher levels of soluble Abeta were not correlated with vulnerability to scopolamine. On the other hand, in aged Tg2576+ mice, cognitive deficits were related to levels of soluble Abeta, not insoluble Abeta, despite the fact that the levels of insoluble Abeta were thousands of times higher than the levels of soluble Abeta. The results of the present experiments suggest that vulnerability to cognitive deficits after scopolamine challenge is not related to elevated levels of soluble Abeta, but that high levels of soluble Abeta are more closely correlated with cognitive deficits than the amount insoluble Abeta, even after large amounts of aggregated, insoluble Abeta have been deposited.

Auditory enhancement of visual temporal order judgment.

Although numerous studies have shown that response times can be speeded by the presentation of multisensory stimuli, here we show that such speeding can be seen even when the second sensory channel fails to provide any task-relevant (i.e. redundant) information, and where cueing appears an unlikely explanation. Study participants performed a visual temporal order judgment task in the presence of task uninformative auditory cues, with the latter sound delayed relative to the latter visual cue. Responses were maximally speeded when the auditory stimulus was delayed by a short time (i.e. 100 ms) relative to the second visual target. These results illustrate a unique form of temporal benefit underlying a multisensory interaction, and form the basis for a novel explanation of these perceptual enhancements.

Effects of CRF1 receptor antagonists and benzodiazepines in the Morris water maze and delayed non-matching to position tests.

RATIONALE: Benzodiazepines continue to be widely used for the treatment of anxiety, but it is well known that benzodiazepines have undesirable side effects, including sedation, ataxia, cognitive deficits and the risk of addiction and abuse. CRF(1) receptor antagonists are being developed as potential novel anxiolytics, but while CRF(1) receptor antagonists seem to have a better side-effect profile than benzodiazepines with respect to sedation and ataxia, the effects of CRF(1) receptor antagonists on cognitive function have not been well characterized. It is somewhat surprising that the potential cognitive effects of CRF(1) receptor antagonists have not been more fully characterized since there is some evidence to suggest that these compounds may impair cognitive function. OBJECTIVE: The Morris water maze and the delayed non-matching to position test are sensitive tests of a range of cognitive functions, including spatial learning, attention and short-term memory, so the objective of the present experiments was to assess the effects of benzodiazepines and CRF(1) receptor antagonists in these tests. RESULTS: The benzodiazepines chlordiazepoxide and alprazolam disrupted performance in the Morris water maze and delayed non-matching to position at doses close to their therapeutic, anxiolytic doses. In contrast, the CRF(1) receptor antagonists DMP-904 and DMP-696 produced little or no impairment in the Morris water maze or delayed non-matching to position test even at doses 10-fold higher than were necessary to produce anxiolytic effects. CONCLUSIONS: The results of the present experiments suggest that, with respect to their effects on cognitive functions, CRF(1) receptor antagonists seem to have a wider therapeutic index than benzodiazepines.

The brain basis of piano performance.

Performances of memorized piano compositions unfold via dynamic integrations of motor, perceptual, cognitive, and emotive operations. The functional neuroanatomy of such elaborately skilled achievements was characterized in the present study by using (15)0-water positron emission tomography to image blindfolded pianists performing a concerto by J.S. Bach. The resulting brain activity was referenced to that for bimanual performance of memorized major scales. Scales and concerto performances both activated primary motor cortex, corresponding somatosensory areas, inferior parietal cortex, supplementary motor area, motor cingulate, bilateral superior and middle temporal cortex, right thalamus, anterior and posterior cerebellum. Regions specifically supporting the concerto performance included superior and middle temporal cortex, planum polare, thalamus, basal ganglia, posterior cerebellum, dorsolateral premotor cortex, right insula, right supplementary motor area, lingual gyrus, and posterior cingulate. Areas specifically implicated in generating and playing scales were posterior cingulate, middle temporal, right middle frontal, and right precuneus cortices, with lesser increases in right hemispheric superior temporal, temporoparietal, fusiform, precuneus, and prefrontal cortices, along with left inferior frontal gyrus. Finally, much greater deactivations were present for playing the concerto than scales. This seems to reflect a deeper attentional focus in which tonically active orienting and evaluative processes, among others, are suspended. This inference is supported by observed deactivations in posterior cingulate, parahippocampus, precuneus, prefrontal, middle temporal, and posterior cerebellar cortices. For each of the foregoing analyses, a distributed set of interacting localized functions is outlined for future test.

Aspects of multisensory perception: the integration of visual and auditory information in musical experiences.

One of the requirements for being a successful musical conductor is to be able to locate sounds instantaneously in time and space. Because this requires the integration of auditory and visual information, the purpose of this study was to examine multisensory processing in conductors and a matched set of control subjects. Subjects participated in a series of behavioral tasks, including pitch discrimination, temporal-order judgment (TOJ), and target localization. Additionally, fMRI scans were done on a subset of subjects who performed a multisensory TOJ task. Analyses of behavioral data indicate that, in the auditory realm, conductors were more accurate in both pitch discrimination and TOJs as well as in locating targets in space. Furthermore, these same subjects also demonstrated a benefit from the combination of auditory and visual information that was not observed in control subjects when locating visual targets. Finally, neural substrates in BA 37, 39/40 were identified as potential areas underlying the conductors' superior multisensory TOJs. Data collection and analyses are ongoing and will lead to an improved understanding of multisensory integration in a complex, musical behavior.

Chronobiotic activity of N-[2-(2,7-dimethoxyfluoren-9-yl)ethyl]-propanamide. Synthesis and melatonergic pharmacology of fluoren-9-ylethyl amides.

A series of fluoren-9-yl ethyl amides (2) were synthesized and evaluated for human melatonin MT(1) and MT(2) receptor binding. N-[2-(2,7-dimethoxyfluoren-9-yl)ethyl]propanamide (2b) was selected and evaluated in functional assays measuring intrinsic activity at the human MT(1) and MT(2) receptors and demonstrated full agonism at both receptors. The chronobiotic properties of 2b were demonstrated in both acute and chronic rat models where 2b produced an acute phase advance of 32 min at 1mg/kg and chronically entrained free-running rats with a mean effective dose of 0.23 mg/kg. Compound 2b is significantly less efficacious than melatonin in constricting human coronary artery.

The song system of the human brain.

Although sophisticated insights have been gained into the neurobiology of singing in songbirds, little comparable knowledge exists for humans, the most complex singers in nature. Human song complexity is evidenced by the capacity to generate both richly structured melodies and coordinated multi-part harmonizations. The present study aimed to elucidate this multi-faceted vocal system by using 15O-water positron emission tomography to scan "listen and respond" performances of amateur musicians either singing repetitions of novel melodies, singing harmonizations with novel melodies, or vocalizing monotonically. Overall, major blood flow increases were seen in the primary and secondary auditory cortices, primary motor cortex, frontal operculum, supplementary motor area, insula, posterior cerebellum, and basal ganglia. Melody repetition and harmonization produced highly similar patterns of activation. However, whereas all three tasks activated secondary auditory cortex (posterior Brodmann Area 22), only melody repetition and harmonization activated the planum polare (BA 38). This result implies that BA 38 is responsible for an even higher level of musical processing than BA 22. Finally, all three of these "listen and respond" tasks activated the frontal operculum (Broca's area), a region involved in cognitive/motor sequence production and imitation, thereby implicating it in musical imitation and vocal learning.