MAP3K8 is a potential therapeutic target in airway epithelial inflammation.

BACKGROUND: We have previously discovered clusters of sequentially negative and positive modulators of acute inflammation during cytokine stimulation in epithelial cells and identified potential targets for therapy within these clusters. MAP3K8 is a druggable kinase that we found to be a hub of a principal interaction network. We describe here the results of MAP3K8 knockdown in the A549 lung cancer cell line, the BEAS-2B epithelial cell line and normal human bronchial epithelial (NHBE) cells following IL-1ß stimulation. We analysed signalling transduction and global gene expression after IL-1ß stimulation with and without MAP3K8 knockdown, quantifying levels of the inflammatory cytokines IL-6, IL-8 and RANTES levels by qPCRs and/or by ELISAs. We also examined potential small molecule inhibitors for MAP3K8 in the same models. RESULTS: IL-1ß significantly and consistently increased MAP3K8 expression after 2 h in A549, BEAS-2B and NHBE cells. Phosphorylation of MAP3K8 occurred at 20 min after IL-1ß stimulation and MAP3K8 protein was degraded at 30 min. MAP3K8 knockdown significantly reduced IL-6, IL-8 levels after IL-1ß stimulation and yielded a 10-fold enhancement of the anti-inflammatory effects of dexamethasone. Phosphorylation of ERK1/2 (P-ERK1/2) and phosphorylation of SAPK/JNK (P-SAPK/JNK) decreased at 30 min after IL-1ß stimulation with MAP3K8 knockdown. The combination of dexamethasone and MAP3K8 knockdown resulted in greater inhibition of phosphorylated ERK1/2 and SAPK/JNK. Nineteen genes including MMP1, MMP3, MMP10, ITGB8, LAMC2 and PLAT (P corrected < 0.01 respectively) demonstrated a distinct altered temporal response to IL-1ß following suppression of MAP3K8. However, putative MAP3K8 inhibitors including Tpl2-1, Tpl2-2 and GSK2222867A only showed inhibition of IL-6 and IL-8 production at a high dose. CONCLUSIONS: These results confirm that MAP3K8 is a key mediator of the early inflammatory response and that it is a potential target in inflammatory diseases. However, current tool compounds do not effectively inhibit its effects.

Comparison of Sub-Ppm Instrument Response Suggests Higher Detection Limits Could Be Used to Quantify Methane Emissions from Oil and Gas Infrastructure.

Quantifying and controlling fugitive methane emissions from oil and gas facilities remains essential for addressing climate goals, but the costs associated with monitoring millions of production sites remain prohibitively expensive. Current thinking, supported by measurement and simple dispersion modelling, assumes single-digit parts-per-million instrumentation is required. To investigate instrument response, the inlets of three trace-methane (sub-ppm) analyzers were collocated on a facility designed to release gas of known composition at known flow rates between 0.4 and 5.2 kg CH4 h-1 from simulated oil and gas infrastructure. Methane mixing ratios were measured by each instrument at 1 Hertz resolution over nine hours. While mixing ratios reported by a cavity ring-down spectrometer (CRDS)-based instrument were on average 10.0 ppm (range 1.8 to 83 ppm), a mid-infrared laser absorption spectroscopy (MIRA)-based instrument reported short-lived mixing ratios far larger than expected (range 1.8 to 779 ppm) with a similar nine-hour average to the CRDS (10.1 ppm). We suggest the peaks detected by the MIRA are likely caused by a micrometeorological phenomenon, where vortex shedding has resulted in heterogeneous methane plumes which only the MIRA can observe. Further analysis suggests an instrument like the MIRA (an optical-cavity-based instrument with cavity size ≤10 cm3 measuring at ≥2 Hz with air flow rates in the order of ≤0.3 slpm at distances of ≤20 m from the source) but with a higher detection limit (25 ppm) could detect enough of the high-concentration events to generate representative 20 min-average methane mixing ratios. Even though development of a lower-cost, high-precision, high-accuracy instrument with a 25 ppm detection threshold remains a significant problem, this has implications for the use of instrumentation with higher detection thresholds, resulting in the reduction in cost to measure methane emissions and providing a mechanism for the widespread deployment of effective leak detection and repair programs for all oil and gas infrastructure.

Ultrasonic vocalisation rate tracks the diurnal pattern of activity in winter phenotype Djungarian hamsters (Phodopus sungorus).

Vocalisations are increasingly being recognised as an important aspect of normal rodent behaviour yet little is known of how they interact with other spontaneous behaviours such as sleep and torpor, particularly in a social setting. We obtained chronic recordings of the vocal behaviour of adult male and female Djungarian hamsters (Phodopus sungorus) housed under short photoperiod (8 h light, 16 h dark, square wave transitions), in different social contexts. The animals were kept in isolation or in same-sex sibling pairs, separated by a grid which allowed non-physical social interaction. On approximately 20% of days hamsters spontaneously entered torpor, a state of metabolic depression that coincides with the rest phase of many small mammal species in response to actual or predicted energy shortages. Animals produced ultrasonic vocalisations (USVs) with a peak frequency of 57 kHz in both social and asocial conditions and there was a high degree of variability in vocalisation rate between subjects. Vocalisation rate was correlated with locomotor activity across the 24-h light cycle, occurring more frequently during the dark period when the hamsters were more active and peaking around light transitions. Solitary-housed animals did not vocalise whilst torpid and animals remained in torpor despite overlapping with vocalisations in social-housing. Besides a minor decrease in peak USV frequency when isolated hamsters were re-paired with their siblings, changing social contexts did not influence vocalisation behaviour or structure. In rare instances, temporally overlapping USVs occurred when animals were socially-housed and were grouped in such a way that could indicate coordination. We did not observe broadband calls (BBCs) contemporaneous with USVs in this paradigm, corroborating their correlation with physical aggression which was absent from our experiment. Overall, we find little evidence to suggest a direct social function of hamster USVs. We conclude that understanding the effects of vocalisations on spontaneous behaviours, such as sleep and torpor, will inform experimental design of future studies, especially where the role of social interactions is investigated.

Tutorial on MUedit: An open-source software for identifying and analysing the discharge timing of motor units from electromyographic signals.

We introduce the open-source software MUedit and we describe its use for identifying the discharge timing of motor units from all types of electromyographic (EMG) signals recorded with multi-channel systems. MUedit performs EMG decomposition using a blind-source separation approach. Following this, users can display the estimated motor unit pulse trains and inspect the accuracy of the automatic detection of discharge times. When necessary, users can correct the automatic detection of discharge times and recalculate the motor unit pulse train with an updated separation vector. Here, we provide an open-source software and a tutorial that guides the user through (i) the parameters and steps of the decomposition algorithm, and (ii) the manual editing of motor unit pulse trains. Further, we provide simulated and experimental EMG signals recorded with grids of surface electrodes and intramuscular electrode arrays to benchmark the performance of MUedit. Finally, we discuss advantages and limitations of the blind-source separation approach for the study of motor unit behaviour during tonic muscle contractions.

Serrated polyposis syndrome: defining the epidemiology and predicting the risk of dysplasia.

BACKGROUND: Serrated polyposis syndrome is the most common polyposis syndrome that has neoplastic potential. However, the natural history, genetic basis, and risk of dysplasia and neoplasia of serrated polyposis syndrome are incompletely understood. The objective of this study is to define the epidemiology of serrated polyposis syndrome. Using this data, we aim to evaluate candidate variables for predicting the risk of dysplasia and neoplasia in sessile serrated lesions found in serrated polyposis syndrome patients. Finally, we aim to use this data to create and evaluate clinical prediction models for accuracy in predicting dysplastic sessile serrated lesions in serrated polyposis syndrome patients. METHODS: This was a regional Australian single-centre retrospective cohort study. Data was prospectively collected data from the clinical record database of a regional Australian gastroenterology practice. All patients undergoing colonoscopy at Port Macquarie Gastroenterology between January 2015 and September 2021 were screened for this study. Collected data included patient demographic, endoscopic, and histopathological findings. Clinical and endoscopic multivariate logistic regression models were created to predict dysplastic sessile serrated lesions. Model performance was examined using the area under the receiver operating curve. RESULTS: In total 8401 patients underwent a colonoscopy procedure during the study period. Serrated polyposis syndrome was diagnosed in 247, representing a prevalence of 2.94% (mean age 67.15 years, 62.75% female). Logistic regression identified; older age at serrated polyposis syndrome diagnosis, a personal history of colorectal cancer, size of the largest sessile serrated lesions removed, and total sessile serrated lesions count as predictors of dysplastic sessile serrated lesions. The clinical and endoscopic model had an area under the receiver operating curve of 0.75. CONCLUSION: Serrated polyposis syndrome is more common than previously described. The clinical and endoscopic variables identified in logistic regression have acceptable accuracy in predicting the risk of dysplasia, however other populations need to be studied to achieve generalisability and improve model performance.

Methods to estimate body temperature and energy expenditure dynamics in fed and fasted laboratory mice: effects of sleep deprivation and light exposure.

Monitoring body temperature and energy expenditure in freely-moving laboratory mice remains a powerful methodology used widely across a variety of disciplines-including circadian biology, sleep research, metabolic phenotyping, and the study of body temperature regulation. Some of the most pronounced changes in body temperature are observed when small heterothermic species reduce their body temperature during daily torpor. Daily torpor is an energy saving strategy characterized by dramatic reductions in body temperature employed by mice and other species when challenged to meet energetic demands. Typical measurements used to describe daily torpor are the measurement of core body temperature and energy expenditure. These approaches can have drawbacks and developing alternatives for these techniques provides options that can be beneficial both from an animal-welfare and study-complexity perspective. First, this paper presents and assesses a method to estimate core body temperature based on measurements of subcutaneous body temperature, and second, a separate approach to better estimate energy expenditure during daily torpor based on core body temperature. Third, the effects of light exposure during the habitual dark phase and sleep deprivation during the light period on body temperature dynamics were tested preliminary in fed and fasted mice. Together, the here-published approaches and datasets can be used in the future to assess body temperature and metabolism in freely-moving laboratory mice.

Recovery of neurophysiological measures in post-COVID fatigue: a 12-month longitudinal follow-up study.

One of the major consequences of the COVID-19 pandemic has been the significant incidence of persistent fatigue following resolution of an acute infection (i.e. post-COVID fatigue). We have shown previously that, in comparison to healthy controls, those suffering from post-COVID fatigue exhibit changes in muscle physiology, cortical circuitry, and autonomic function. Whether these changes preceded infection, potentially predisposing people to developing post-COVID fatigue, or whether the changes were a consequence of infection was unclear. Here we present results of a 12-month longitudinal study of 18 participants from the same cohort of post-COVID fatigue sufferers to investigate these correlates of fatigue over time. We report improvements in self-perception of the impact of fatigue via questionnaires, as well as significant improvements in objective measures of peripheral muscle fatigue and autonomic function, bringing them closer to healthy controls. Additionally, we found reductions in muscle twitch tension rise times, becoming faster than controls, suggesting that the improvement in muscle fatigability might be due to a process of adaptation rather than simply a return to baseline function.

Combination of Extended Antivirals With Antiretrovirals for Severe Mpox in Advanced Human Immunodeficiency Virus Infection: Case Series of 4 Patients.

To gauge the safety and utility of extended tecovirimat/cidofovir for severe mpox, here we report our experience caring for 4 patients with mpox and advanced human immunodeficiency virus (HIV) at the Hospitals of the University of Pennsylvania during the 2022 global outbreak. Three patients had recurrent courses complicated by superinfections, coinfections and insufficient nutrition/housing, requiring extended tecovirimat (5-16 weeks) and cidofovir (1-12 doses) with probenecid and fluids. At follow-up, patients had undetectable HIV RNA on antiretrovirals, improved ulcers and stable renal function on antivirals. Serology guided cessation for one 7-month cidofovir course. Overall findings support a comprehensive approach of prolonged tecovirimat/cidofovir with antiretrovirals for severe mpox, while addressing social factors.

Recommendations for measuring and standardizing light for laboratory mammals to improve welfare and reproducibility in animal research.

Light enables vision and exerts widespread effects on physiology and behavior, including regulating circadian rhythms, sleep, hormone synthesis, affective state, and cognitive processes. Appropriate lighting in animal facilities may support welfare and ensure that animals enter experiments in an appropriate physiological and behavioral state. Furthermore, proper consideration of light during experimentation is important both when it is explicitly employed as an independent variable and as a general feature of the environment. This Consensus View discusses metrics to use for the quantification of light appropriate for nonhuman mammals and their application to improve animal welfare and the quality of animal research. It provides methods for measuring these metrics, practical guidance for their implementation in husbandry and experimentation, and quantitative guidance on appropriate light exposure for laboratory mammals. The guidance provided has the potential to improve data quality and contribute to reduction and refinement, helping to ensure more ethical animal use.

Supramolecular Control of the Temperature Responsiveness of Fluorescent Macrocyclic Molecular Rotamers.

To fully harness the potential of molecular machines, it is crucial to develop methods by which to exert control over their speed of motion through the application of external stimuli. A conformationally strained macrocyclic fluorescent rotamer, CarROT, displays a reproducible and linear fluorescence decrease towards temperature over the physiological temperature range. Through the external addition of anions, cations or through deprotonation, the compound can access four discreet rotational speeds via supramolecular interactions (very slow, slow, fast and very fast) which in turn stop, reduce or enhance the thermoluminescent properties due to increasing or decreasing non-radiative decay processes, thereby providing a means to externally control the temperature sensitivity of the system. Through comparison with analogues with a higher degree of conformational freedom, the high thermosensitivity of CarROT over the physiological temperature range was determined to be due to conformational strain, which causes a high energy barrier to rotation over this range. Analogues with a higher degree of conformational freedom display lower sensitivities towards temperature over the same temperature range. This study provides an example of an information rich small molecule, in which programable rotational speed states can be observed with facile read-out.

Methane emissions from abandoned oil and gas wells in Colorado.

Recent studies indicate emission factors used to generate bottom-up methane inventories may have considerable regional variability. The US's Environmental Protection Agency's emission factors for plugged and unplugged abandoned oil and gas wells are largely based on measurement of historic wells and estimated at 0.4 g and 31 g CH4 well-1 h-1, respectively. To investigate if these are representative of wells more recently abandoned, methane emissions were measured from 128 plugged and 206 unplugged abandoned wells in Colorado, finding the first super-emitting abandoned well (76 kg CH4 well-1 h-1) and average emissions of 0 and 586 g CH4 well-1 h-1, respectively. Combining these with other states' measurements, we update the US emission factors to 1 and 198 g CH4 well-1 h-1, respectively. Correspondingly, annual methane emissions from the 3.4 million abandoned wells in the US are estimated at between 2.6 Tg, following current methodology, and 1.1 Tg, where emissions are disaggregated for well-type. In conclusion, this study identifies a new abandoned well-type, recently-producing orphaned, that contributes 74 % to the total abandoned wells methane emissions. Including this new well-type in the bottom-up inventory suggests abandoned well emissions equate to between 22 and 49 % of total emissions from US active oil and gas production operations.

Quantifying non-steady state natural gas leakage from the pipelines using an innovative sensor network and model for subsurface emissions - InSENSE.

Detecting and quantifying subsurface leaks remains a challenge due to the complex nature and extent of belowground leak scenarios. To address these scenarios, monitoring and evaluating changes in gas leakage behavior over space and time are crucial for ensuring safe and efficient responses to known or potential gas leaks. This study demonstrates the capability of linking environmental and gas concentration data obtained using a low-cost, near real-time methane (CH4) detector network and an inverse gas migration model to capture and quantify non-steady state belowground natural gas (NG) leaks. The Estimating Surface Concentration Above Pipeline Emission (ESCAPE) model was modified to incorporate the impact of soil properties on gas migration. Field-scale controlled NG experiments with leakage rates ranging from 37 to 121 g/h indicate that elevated belowground near-surface (BNS) gas concentrations persist long before elevated surface concentrations are observed. On average, BNS CH4 concentrations were 20%-486% higher than surface CH4 concentrations within the monitoring radius of 4 m from the leak location. An increase in the BNS CH4 concentration was observed within 3 h as the leak rate increased from 37 to 89 g/h. However, due to the atmospheric fluctuations, any changes in surface CH4 concentrations could not be confirmed within this period. The plume area of the BNS CH4 extended approximately two times farther than that of the surface CH4 as the gas leak rate increased from 37 to 121 g/h. The estimated NG leak rates by the modified ESCAPE model agreed well with the experimental NG leak rates (m = 0.99 and R2 = 0.77), demonstrating that including soil characteristics and BNS CH4 measurements can advance estimations of non-steady NG leak rates in low and moderate NG leak rate scenarios. The CH4 detector network and model show potential as an innovative tool to improve operators' risk assessment and NG leakage response.

Uncertainty Quantification of Methods Used to Measure Methane Emissions of 1 g CH4 h-1.

The recent interest in measuring methane (CH4) emissions from abandoned oil and gas wells has resulted in five methods being typically used. In line with the US Federal Orphaned Wells Program's (FOWP) guidelines and the American Carbon Registry's (ACR) protocols, quantification methods must be able to measure minimum emissions of 1 g of CH4 h-1 to within ±20%. To investigate if the methods meet the required standard, dynamic chambers, a Hi-Flow (HF) sampler, and a Gaussian plume (GP)-based approach were all used to quantify a controlled emission (Qav; g h-1) of 1 g of CH4 h-1. After triplicate experiments, the average accuracy (Ar; %) and the upper (Uu; %) and lower (Ul; %) uncertainty bounds of all methods were calculated. Two dynamic chambers were used, one following the ACR guidelines, and a second "mobile" chamber made from lightweight materials that could be constructed around a source of emission on a well head. The average emission calculated from the measurements made using the dynamic chamber (Qav = 1.01 g CH4 h-1, Ar = +0.9%), the mobile chamber (Qav = 0.99 g CH4 h-1, Ar = -1.4%), the GP approach (Qav = 0.97 g CH4 h-1, Ar = -2.6%), and the HF sampler (Qav = 1.02 g CH4 h-1, Ar = +2.2%) were all within ±3% of 1 g of CH4 h-1 and met the requirements of the FOWP and ACR protocols. The results also suggest that the individual measurements made using the dynamic chamber can quantify emissions of 1 g of CH4 h-1 to within ±6% irrespective of the design (material, number of parts, geometrical shape, and hose length), and changes to the construction or material specifications as defined via ACR make no discernible difference to the quantification uncertainty. Our tests show that a collapsible chamber can be easily constructed around the emission source on an abandoned well and be used to quantify emissions from abandoned wells in remote areas. To our knowledge, this is the first time that methods for measuring the CH4 emissions of 1 g of CH4 h-1 have been quantitively assessed against a known reference source and against each other.

Dynamic and Isometric Force-Time Curve Characteristics Influencing Change of Direction Performance of State-Level Netball Players.

ABSTRACT: Smolarek, T, Haff, GG, Poon, WCK, Nagatani, T, Barley, OR, and Guppy, SN. Dynamic and isometric force-time curve characteristics influencing change of direction performance of state-level netball players. J Strength Cond Res 37(12): 2397-2404, 2023-Although multiple lower-body strength capacities are required to change direction rapidly, there is limited knowledge about the relative importance of these factors. Therefore, the purpose of this study was to assess the relationship between dynamic and isometric lower-body strength capacities and change of direction (COD) time in state-league netball players. Seventeen female athletes completed maximal isometric midthigh pull (IMTP), countermovement jump (CMJ), and modified 5-0-5 COD tests. Pearson's product moment correlations were used to determine the relationship between COD time and several IMTP and CMJ force-time curve characteristics. To assess the level of contribution of each force-time curve characteristic to COD time, multivariate-linear stepwise regression analyses were performed. A significant moderate correlation was noted between net relative peak force (PF) during the IMTP and COD time ( r = 0.488, p = 0.047), accounting for 23.8% of the variance in COD time. Moreover, concentric relative impulse during the CMJ was strongly correlated with COD time ( r = 0.718; p = 0.001), explaining 81.9% of the variance in COD time when combined with net relative braking PF in a stepwise regression. Based on these findings, female netball players who display higher concentric and isometric strength, as well as the ability to express higher impulses during the concentric phase of the CMJ, are likely to perform CODs faster. This may occur because COD requires the generation of greater propulsive forces, as well as reduced braking and contact times, along with greater isometric strength enabling effective repositioning of center of mass during COD tasks.

A macrocyclic fluorescent probe for the detection of citrate.

We present a macrocyclic fluorescent probe for the detection of citrate. This receptor binds citrate through hydrogen-bonding interactions in aqueous solutions, and exhibits a turn-on in fluorescence in response to binding. The presence of common biologically relevant dicarboxylate species does not significantly impact the fluorescence response. We have demonstrated the utility of this probe with the staining of murine splenocytes, and identified different basal levels of citrate present in immune cell subsets via flow cytometry analysis.

Estimating the Below-Ground Leak Rate of a Natural Gas Pipeline Using Above-Ground Downwind Measurements: The ESCAPE-1 Model.

Natural gas (NG) leaks from below-ground pipelines pose safety, economic, and environmental hazards. Despite walking surveys using handheld methane (CH4) detectors to locate leaks, accurately triaging the severity of a leak remains challenging. It is currently unclear whether CH4 detectors used in walking surveys could be used to identify large leaks that require an immediate response. To explore this, we used above-ground downwind CH4 concentration measurements made during controlled emission experiments over a range of environmental conditions. These data were then used as the input to a novel modeling framework, the ESCAPE-1 model, to estimate the below-ground leak rates. Using 10-minute averaged CH4 mixing/meteorological data and filtering out wind speed < 2 m s-1/unstable atmospheric data, the ESCAPE-1 model estimates small leaks (0.2 kg CH4 h-1) and medium leaks (0.8 kg CH4 h-1) with a bias of -85%/+100% and -50%/+64%, respectively. Longer averaging (≥3 h) results in a 55% overestimation for small leaks and a 6% underestimation for medium leaks. These results suggest that as the wind speed increases or the atmosphere becomes more stable, the accuracy and precision of the leak rate calculated by the ESCAPE-1 model decrease. With an uncertainty of ±55%, our results show that CH4 mixing ratios measured using industry-standard detectors could be used to prioritize leak repairs.

Mpox: Keep it on the differential.

In its current global outbreak, mpox has exhibited several novel clinical presentations that clinicians should be aware of so they can recognize it if they see it. Although the case rate has decreased, mpox could linger at a low rate or resurface in other populations and thus should remain in the differential diagnosis in patients presenting with potential infections after intimate encounters.

Effect of 3 Different Set Configurations on Kinematic Variables and Internal Loads During a Power Snatch Session.

ABSTRACT: Nagatani, T, Kendall, KL, Guppy, SN, Poon, WCK, and Haff, GG. Effect of 3 different set configurations on kinematic variables and internal loads during a power snatch session. J Strength Cond Res 37(10): 1929-1938, 2023-The aim of this study was to investigate the effect of 3 different set configurations on kinematic variables and internal loads during multiple sets performed with the power snatch. Ten strength-power athletes with at least 6 months of training experience performing the power snatch participated in this study, which consisted of 3 experimental protocols performed in a randomized repeated-measures design. The 3 protocols involved performing the power snatch for 3 sets of 5 repetitions at an average load of 75% 1 repetition maximum with a traditional (TRAD), cluster (CLU), or ascending cluster (A-CLU) protocol, where the training load was progressively increased across the set. Kinematic variables and internal loads (heart rate, blood lactate, and rate of perceived exertion) were measured during each protocol. The athletes maintained peak velocity (PV) and peak power (PP) and exhibited lower internal loads during CLU sets when compared with TRAD sets, whereas they displayed significant decreases in PV during TRAD sets. However, there were no statistically significant differences in PV and PP responses between the TRAD and CLU protocol. The athletes exhibited a significant decrease in PV, whereas PP was increased across each set in the A-CLU protocol, with lower internal loads observed compared with the TRAD protocol. Overall, the training loads used in this study do not appear to maximize the benefits of using CLU set during 3 sets of power snatches performed for 5 repetitions. In addition, A-CLU sets may potentially be useful as a means of maximizing the power output of the athlete.

Continuous home cage monitoring of activity and sleep in mice during repeated paroxetine treatment and discontinuation.

RATIONALE: Non-invasive home cage monitoring is emerging as a valuable tool to assess the effects of experimental interventions on mouse behaviour. A field in which these techniques may prove useful is the study of repeated selective serotonin reuptake inhibitor (SSRI) treatment and discontinuation. SSRI discontinuation syndrome is an under-researched condition that includes the emergence of sleep disturbances following treatment cessation. OBJECTIVES: We used passive infrared (PIR) monitoring to investigate changes in activity, sleep, and circadian rhythms during repeated treatment with the SSRI paroxetine and its discontinuation in mice. METHODS: Male mice received paroxetine (10 mg/kg/day, s.c.) for 12 days, then were swapped to saline injections for a 13 day discontinuation period and compared to mice that received saline injections throughout. Mice were continuously tracked using the Continuous Open Mouse Phenotyping of Activity and Sleep Status (COMPASS) system. RESULTS: Repeated paroxetine treatment reduced activity and increased behaviourally-defined sleep in the dark phase. These effects recovered to saline-control levels within 24 h of paroxetine cessation, yet there was also evidence of a lengthening of sleep bouts in the dark phase for up to a week following discontinuation. CONCLUSIONS: This study provides the first example of how continuous non-invasive home cage monitoring can be used to detect objective behavioural changes in activity and sleep during and after drug treatment in mice. These data suggest that effects of paroxetine administration reversed soon after its discontinuation but identified an emergent change in sleep bout duration, which could be used as a biomarker in future preclinical studies to prevent or minimise SSRI discontinuation symptoms.