Dynamic behaviour restructuring mediates dopamine-dependent credit assignment.

Animals exhibit a diverse behavioural repertoire when exploring new environments and can learn which actions or action sequences produce positive outcomes. Dopamine release after encountering a reward is critical for reinforcing reward-producing actions1-3. However, it has been challenging to understand how credit is assigned to the exact action that produced the dopamine release during continuous behaviour. Here we investigated this problem in mice using a self-stimulation paradigm in which specific spontaneous movements triggered optogenetic stimulation of dopaminergic neurons. Dopamine self-stimulation rapidly and dynamically changes the structure of the entire behavioural repertoire. Initial stimulations reinforced not only the stimulation-producing target action, but also actions similar to the target action and actions that occurred a few seconds before stimulation. Repeated pairings led to a gradual refinement of the behavioural repertoire to home in on the target action. Reinforcement of action sequences revealed further temporal dependencies of refinement. Action pairs spontaneously separated by long time intervals promoted a stepwise credit assignment, with early refinement of actions most proximal to stimulation and subsequent refinement of more distal actions. Thus, a retrospective reinforcement mechanism promotes not only reinforcement, but also gradual refinement of the entire behavioural repertoire to assign credit to specific actions and action sequences that lead to dopamine release.

The experiences of patients using a cancer hotline service.

BACKGROUND: The present study aimed to evaluate oncology patients' experiences with a hotline service in a tertiary cancer centre, identifying areas for local improvement and to inform the work of the hotline. METHODS: A link to an online platform to complete a survey was sent through bulk messaging to 3028 mobile numbers. The survey answers and results were saved and consolidated in the online platform. FINDINGS: The survey received 368 responses: 49% of participants had their calls answered within 10 minutes, while 18% (n=66) waited 30 minutes or more. Eighty-two per cent were satisfied with the length of time they waited to speak with a nurse and 71% reported that their overall experience was very good. Ninety-seven per cent of participants felt listened to and were offered support and 91% felt that their reason for calling was adequately addressed. CONCLUSION: The present study offers persuasive evidence indicating that oncology patients are largely satisfied with the hotline service; nonetheless, it is recognised that a more robust evaluation is needed. Regardless, the need for certain improvements has been identified to offer the potential to enhance patients' experience.

Association between Operative Time and Short-Term Radical Cystectomy Complications.

INTRODUCTION: The aim of this study was to examine the relationship between duration of surgical intervention and postoperative complications in radical cystectomy (RC). We hypothesized that the complication rate increases with longer operative time. METHODS: We analyzed the National Surgical Quality Improvement Program database 2011-2017 to identify all patients who underwent RC. Clinicodemographic characteristics, operative time, and perioperative complications using the Clavien-Dindo Classification (CDC) were abstracted. We fit a generalized linear model with linear splines for operative time to analyze if the relationship between operative time and probability of complication changed over time. RESULTS: A total of 10,520 RC patients were identified with a mean operative time of 5.5 h (standard deviation 2.03). In 55% and 18.2%, any complication and major complications (CDC ≥3) occurred within 30 days postoperatively, respectively. The spline regression model for any complication showed an almost linear relationship between the complication rate and operative time, ranging from 55% at 2.5 h to 82% at 10 h. For major complications, the model revealed the inflection point (knot) at 4.5 h, which corresponds to the lowest complication rate with 15%. Operative times at the extremes of the distribution had higher complication rates: 17.5% if <2.5 h and 28% if >10 h. DISCUSSION/CONCLUSION: Operative time of RC is associated with postoperative complications. Though many factors impact the duration of surgery, surgeries that lasted between 4 and 5 h had trend toward the lowest complication rates. Attention to factors impacting operative time may allow surgeons to identify strategies for optimizing surgical care and reducing complications after RC.

Development of a Biomechanical Device for Parameter Quantification Associated with the Sit-to-Stand Movement.

The "sit-to-stand" (STS) movement is essential during activities of daily living (ADL). In individuals with physical-motor diseases, its execution and repetition increases activity levels, which is crucial for a good motor rehabilitation process and daily training. Interestingly, there are no sit-to-stand devices that allow a quantitative assessment of the key variables that happen during STS, and there is a need to come up with a new device. This work presents a developed biomechanical support device that measures the force of the upper limbs during the STS movement, aiming to motivate and encourage people undergoing physical therapy in the lower limbs. The device uses two instrumented beams and allows real-time visualization of both arms' applied force and it records the data for post-processing. The device was tested with a well-defined protocol on a group of 34 healthy young volunteers and an elderly group of 16 volunteers from a continuing care unit. The system showed robust strength and stiffness, good usability, and a user interface that acquired and recorded data effectively, allowing one to observe force-time during the execution of the movement through the application interface developed and in recording data for post-processing. Asymmetries in the applied forces in the STS movement between the upper limbs were identified, particularly in volunteers of the continuing care unit. From the application and the registered data, it can be observed that volunteers with motor problems in the lower limbs performed more strength in their arms to compensate. As expected, the maximum average strength of the healthy volunteers (both arms: force = 105 Newton) was higher than that of the volunteers from the continuing care unit (right arm: force = 54 Newton; left arm: force = 56 Newton). Among others, moderate correlations were observed between weight-applied and height-applied forces and there was a moderately high correlation between the Sequential Clinical Assessment of Respiratory Function (SCAR-F score) and time to perform the movement. Based on the obtained results, the developed device can be a helpful tool for monitoring the evaluation of a patient with limitations in the upper and lower limbs. In addition, the developed system allows for easy evolution, such as including a barometric platform and implementing serious games that can stimulate the execution of the STS movement.

Active surveillance for intermediate-risk prostate cancer.

INTRODUCTION: Active surveillance (AS) is an established approach in the management of low-risk, localized prostate cancer. While the use of AS to manage intermediate-risk (IR) disease has gradually increased over time, there remains uncertainty with regards to its safety, with only a minority of IR patients currently being managed with this approach. MATERIALS AND METHODS: We conducted a narrative review based on an analysis of the literature focusing on articles describing AS for IR prostate cancer. We focus on the uncertainty surrounding AS in IR disease by discussing variations in the definitions and guideline recommendations associated with IR disease, and describing the limitations of the evidence from observational studies and randomized trials. CONCLUSION: The safety of AS for IR disease remains unknown, given the lack of randomized trials and the limitations of the current observational studies. Further research is needed to identify select patients with IR prostate cancer that can be managed safely with AS.

Effects of roads on small-mammal movements: Opportunities and risks of vegetation management on roadsides.

Roads can block animal movement and reduce persistence of species living in road surroundings. Movement restrictions on local populations may even increase extinction risk of abundant small mammals. However, road verges (road managed area between the edge of the road and the beginning of private land) may provide refuge and corridors for small mammals when properly managed. Information on the effects of roads and roadside management on small-mammal movement is still scarce for low traffic roads (<20,000 vehicles per day) crossing well-preserved habitats. We aimed to fill this gap by comparing fine-scale movement patterns of wood mice (Apodemus sylvaticus) in a road and in a similar roadless area without management. Both areas consisted of a well-preserved Mediterranean agro-silvo pastoral system. We studied several movement patterns: road crossings, verge use, length, and direction of movement. Additionally, we assessed how roadside management, animals' sex and residency status, season and microhabitat affect movement at the road area. At the roadless area, we defined a virtual road and verges at equivalent locations to the road area for comparison purposes. We gathered capture-mark-recapture data for two years to characterize movement patterns. Wood mice tended to avoid the road by crossing it less often and moving away from it more frequently than from equivalent locations in the roadless area. Wood mice used road verges more frequently than virtual verges and moved more often parallel to the road than to the virtual road. Road crossings were more frequent after firebreak openings (strips of mowed land) in surrounding areas and near taller shrubs. Also, males used road verges more often than females. Differences on several movement patterns between areas and their trends within the road area can be explained mainly by the presence of the road and roadside vegetation management (e.g., firebreaks openings). We suggest roadside vegetation management practices (e.g., avoid land mowing; maintain vegetation strips) to promote the role of verges as refuges and/or corridors for small mammals.

Advanced nursing practice: a review of scopes of practice in cancer care.

The establishment of advanced nurse practitioners (ANPs) has expanded considerably in recent years and shown to result in substantial contributions to numerous fields of health care. Due to advancements in treatments and innovations in medicine, patients with cancer are living longer, requiring a multifactorial holistic approach in which ANPs, due to their skills and knowledge, can be best utilised, as they are able to provide the expert care required at various stages of the patient journey. This article explores scopes of practice from ANPs working with oncology patients in a tertiary cancer centre, making explicit their roles, in addition to highlighting experienced challenges and future directions of care.

Predicting survival after radical prostatectomy: Variation of machine learning performance by race.

BACKGROUND: Robust prediction of survival can facilitate clinical decision-making and patient counselling. Non-Caucasian males are underrepresented in most prostate cancer databases. We evaluated the variation in performance of a machine learning (ML) algorithm trained to predict survival after radical prostatectomy in race subgroups. METHODS: We used the National Cancer Database (NCDB) to identify patients undergoing radical prostatectomy between 2004 and 2016. We grouped patients by race into Caucasian, African-American, or non-Caucasian, non-African-American (NCNAA) subgroups. We trained an Extreme Gradient Boosting (XGBoost) classifier to predict 5-year survival in different training samples: naturally race-imbalanced, race-specific, and synthetically race-balanced. We evaluated performance in the test sets. RESULTS: A total of 68,630 patients met inclusion criteria. Of these, 57,635 (84%) were Caucasian, 8173 (12%) were African-American, and 2822 (4%) were NCNAA. For the classifier trained in the naturally race-imbalanced sample, the F1 scores were 0.514 (95% confidence interval: 0.513-0.511), 0.511 (0.511-0.512), 0.545 (0.541-0.548), and 0.378 (0.378-0.389) in the race-imbalanced, Caucasian, African-American, and NCNAA test samples, respectively. For all race subgroups, the F1 scores of classifiers trained in the race-specific or synthetically race-balanced samples demonstrated similar performance compared to training in the naturally race-imbalanced sample. CONCLUSIONS: A ML algorithm trained using NCDB data to predict survival after radical prostatectomy demonstrates variation in performance by race, regardless of whether the algorithm is trained in a naturally race-imbalanced, race-specific, or synthetically race-balanced sample. These results emphasize the importance of thoroughly evaluating ML algorithms in race subgroups before clinical deployment to avoid potential disparities in care.

Cynotherapy in Cancer Pain Management: A Pilot Study.

OBJECTIVES: This was a pilot study to evaluate the feasibility and impact of a single dog-assisted therapy (cynotherapy) session in reducing pain and emotional distress in oncological outpatients compared with typical waiting room experience (control). DESIGN: This was a quasi-experimental before-after controlled study that took place at a chronic pain outpatient clinic of a tertiary cancer center, whose participants were adult oncological patients, able to consent and without medical contraindication. SETTING: Chronic pain outpatient clinic of a tertiary cancer center. PARTICIPANTS: Adult oncological patients able to consent and without medical contraindication. METHODS: All participants completed self-reported questionnaires including a numeric rating scale for pain and distress thermometer at admission and immediately before departure from the clinic. RESULTS: Eighty-one patients were enrolled over a 10-month study period, 41 in the cynotherapy group and 40 controls. Improvement was greater in cynotherapy than control group for pain (median difference score = -1.0 vs 0.0; P = 0.037), distress levels (median = -1.0 vs 0.0; P = 0.017), and depression (median = -1.0 vs 0.0; P = 0.030). The proportion of patients with a clinically relevant improvement in pain (reduction ≥2 points) was approximately twofold in the cynotherapy group when compared with controls, although not statistically significant (39% vs 20%, odds ratio = 2.53, 95% confidence interval = 0.86-8.02; P = 0.088). The mean satisfaction rate was 9.3/10, and no negative occurrences were reported. CONCLUSION: A single session of dog-assisted therapy can provide immediate improvement in the perception of pain and distress for patients with chronic cancer pain in an outpatient setting, with high satisfaction rates and no negative occurrences. This nonrandomized pilot study points toward the clinical relevance of implementing cynotherapy at a cancer pain clinic and developing a larger scale, more directed study.

HflXr, a homolog of a ribosome-splitting factor, mediates antibiotic resistance.

To overcome the action of antibiotics, bacteria have evolved a variety of different strategies, such as drug modification, target mutation, and efflux pumps. Recently, we performed a genome-wide analysis of Listeria monocytogenes gene expression after growth in the presence of antibiotics, identifying genes that are up-regulated upon antibiotic treatment. One of them, lmo0762, is a homolog of hflX, which encodes a heat shock protein that rescues stalled ribosomes by separating their two subunits. To our knowledge, ribosome splitting has never been described as an antibiotic resistance mechanism. We thus investigated the role of lmo0762 in antibiotic resistance. First, we demonstrated that lmo0762 is an antibiotic resistance gene that confers protection against lincomycin and erythromycin, and that we renamed hflXr (hflX resistance). We show that hflXr expression is regulated by a transcription attenuation mechanism relying on the presence of alternative RNA structures and a small ORF encoding a 14 amino acid peptide containing the RLR motif, characteristic of macrolide resistance genes. We also provide evidence that HflXr is involved in ribosome recycling in presence of antibiotics. Interestingly, L. monocytogenes possesses another copy of hflX, lmo1296, that is not involved in antibiotic resistance. Phylogenetic analysis shows several events of hflXr duplication in prokaryotes and widespread presence of hflXr in Firmicutes. Overall, this study reveals the Listeria hflXr as the founding member of a family of antibiotic resistance genes. The resistance conferred by this gene is probably of importance in the environment and within microbial communities.

Comprehensive Assessment of Endomyocardial Fibrosis with Cardiac MRI: Morphology, Function, and Tissue Characterization.

Endomyocardial fibrosis (EMF) affects approximately 12 million persons worldwide and is an important cause of restrictive cardiomyopathy in the developing world, with the highest prevalence reported in sub-Saharan Africa, South Asia, and South America. EMF is characterized by apical filling with fibrotic tissue of one or both ventricles, often associated with thrombus, calcification, and atrioventricular valve regurgitation, leading to typical symptoms of restrictive heart failure. Transthoracic echocardiography (TTE) is the first-line modality for assessment of EMF, basically owing to its widespread availability. However, in recent years cardiac MRI has emerged as a powerful tool for assessment of cardiac morphology and function, with higher accuracy than TTE, along with the unique advantage of being able to provide comprehensive noninvasive tissue characterization. Delayed enhancement (DE) imaging is the cornerstone of cardiac MRI tissue characterization and allows accurate identification of myocardial fibrosis, conveying valuable additional diagnostic and prognostic information. The typical DE pattern in EMF, described as the "double V" sign, consists of a three-layered pattern of normal myocardium, thickened enhanced endomyocardium, and overlying thrombus at the apex of the affected ventricle; it has excellent correlation with histopathologic findings and plays an important role in differentiating EMF from other cardiomyopathies. Conversely, fibrous tissue deposition quantified using DE imaging, when indexed to body surface area, has been shown to be a strong independent predictor of mortality. The aim of this review is to summarize state-of-the-art applications of cardiac MRI for diagnostic and prognostic assessment of patients with suspected or confirmed EMF. Online supplemental material is available for this article. ©RSNA, 2020.

Stathmin recruits tubulin to Listeria monocytogenes-induced actin comets and promotes bacterial dissemination.

The tubulin cytoskeleton is one of the main components of the cytoarchitecture and is involved in several cellular functions. Here, we examine the interplay between Listeria monocytogenes (Lm) and the tubulin cytoskeleton upon cellular infection. We show that non-polymeric tubulin is present throughout Lm actin comet tails and, to a less extent, in actin clouds. Moreover, we demonstrate that stathmin, a regulator of microtubule dynamics, is also found in these Lm-associated actin structures and is required for tubulin recruitment. Depletion of host stathmin results in longer comets containing less F-actin, which may be correlated with higher levels of inactive cofilin in the comet, thus suggesting a defect on local F-actin dynamics. In addition, intracellular bacterial speed is significantly reduced in stathmin-depleted cells, revealing the importance of stathmin/tubulin in intracellular Lm motility. In agreement, the area of infection foci and the total bacterial loads are also significantly reduced in stathmin-depleted cells. Collectively, our results demonstrate that stathmin promotes efficient cellular infection, possibly through tubulin recruitment and control of actin dynamics at Lm-polymerized actin structures.

Rapid Remodeling of the Host Epithelial Cell Proteome by the Listeriolysin O (LLO) Pore-forming Toxin.

Bacterial pathogens use various strategies to interfere with host cell functions. Among these strategies, bacteria modulate host gene transcription, thereby modifying the set of proteins synthetized by the infected cell. Bacteria can also target pre-existing host proteins and modulate their post-translational modifications or trigger their degradation. Analysis of protein levels variations in host cells during infection allows to integrate both transcriptional and post-transcriptional regulations induced by pathogens. Here, we focused on host proteome alterations induced by the toxin Listeriolysin O (LLO), secreted by the bacterial pathogen Listeria monocytogenes. We showed that a short-term treatment with LLO remodels the host cell proteome by specifically decreasing the abundance of 149 proteins. The same decrease in host protein levels was observed in different epithelial cell lines but not in macrophages. We show in particular that this proteome remodeling affects several ubiquitin and ubiquitin-like ligases and that LLO leads to major changes in the host ubiquitylome. Strikingly, this toxin-induced proteome remodeling involves only post-transcriptional regulations, as no modification in the transcription levels of the corresponding genes was observed. In addition, we could show that Perfringolysin O, another bacterial pore-forming toxin similar to LLO, also induces host proteome changes. Taken together, our data reveal that different bacterial pore-forming toxins induce important host proteome remodeling, that may impair epithelial cell functions.

MouR controls the expression of the Listeria monocytogenes Agr system and mediates virulence.

The foodborne pathogen Listeria monocytogenes (Lm) causes invasive infection in susceptible animals and humans. To survive and proliferate within hosts, this facultative intracellular pathogen tightly coordinates the expression of a complex regulatory network that controls the expression of virulence factors. Here, we identified and characterized MouR, a novel virulence regulator of Lm. Through RNA-seq transcriptomic analysis, we determined the MouR regulon and demonstrated how MouR positively controls the expression of the Agr quorum sensing system (agrBDCA) of Lm. The MouR three-dimensional structure revealed a dimeric DNA-binding transcription factor belonging to the VanR class of the GntR superfamily of regulatory proteins. We also showed that by directly binding to the agr promoter region, MouR ultimately modulates chitinase activity and biofilm formation. Importantly, we demonstrated by in vitro cell invasion assays and in vivo mice infections the role of MouR in Lm virulence.

Road verges provide connectivity for small mammals: A case study with wood mice (Apodemus sylvaticus) in an agro-silvo pastoral system.

Roads disrupt landscape connectivity for many terrestrial mammals. These infrastructures can be barriers to movement thereby threatening population persistence. Nonetheless, small mammals may use road verges as habitat or corridor, thus increasing migration across intensively managed landscapes. However, in well-preserved habitats where road verges show a similar vegetation structure to surrounding areas, their role is still unknown. Road verges would have an important role as fine-scale connectivity providers for small mammals in a well-preserved habitat depending on land management on road surroundings. We aimed to quantify the effects of road verges and paved lanes on the fine-scale landscape connectivity for the wood mouse (Apodemus sylvaticus) in a well-preserved Mediterranean woodland. Additionally, we assessed the impact on connectivity of vegetation cutting on verges and of management in surrounding areas (i.e. firebreaks, grazing, ploughing and cork stripping). We quantified connectivity using graph theory based on two years of capture-recapture data. We compared a set of connectivity metrics (derived from the probability of connectivity index) in a road area and in a virtual roadless scenario. We found that the presence of the road reduced overall fine-scale landscape connectivity, acting as a partial barrier for wood mice movement. However, verges had a key role in promoting movement on road surroundings. Vegetation cutting on verges, and land ploughing in the surrounding landscape were the only management activities compromising connectivity. Our study supports the already known role of road verges as habitat corridors for small mammals. However, it goes beyond existing knowledge by quantifying the connectivity enhancement provided by road verges and demonstrating that this role is highly relevant even in well-preserved landscapes. Therefore, our findings emphasize the critical role of road verges and suggest important management options to enhance landscape connectivity for small mammals.

Altered bone microarchitecture in a type 1 diabetes mouse model Ins2Akita.

Type 1 diabetes mellitus (T1DM) has been associated to several cartilage and bone alterations including growth retardation, increased fracture risk, and bone loss. To determine the effect of long term diabetes on bone we used adult and aging Ins2 Akita mice that developed T1DM around 3-4 weeks after birth. Both Ins2 Akita and wild-type (WT) mice were analyzed at 4, 6, and 12 months to assess bone parameters such as femur length, growth plate thickness and number of mature and preapoptotic chondrocytes. In addition, bone microarchitecture of the cortical and trabecular regions was measured by microcomputed tomography and gene expression of Adamst-5, Col2, Igf1, Runx2, Acp5, and Oc was quantified by quantitative real-time polymerase chain reaction. Ins2 Akita mice showed a decreased longitudinal growth of the femur that was related to decreased growth plate thickness, lower number of chondrocytes and to a higher number of preapoptotic cells. These changes were associated with higher expression of Adamst-5, suggesting higher cartilage degradation, and with low expression levels of Igf1 and Col2 that reflect the decreased growth ability of diabetic mice. Ins2 Akita bone morphology was characterized by low cortical bone area (Ct.Ar) but higher trabecular bone volume (BV/TV) and expression analysis showed a downregulation of bone markers Acp5, Oc, and Runx2. Serum levels of insulin and leptin were found to be reduced at all-time points Ins2 Akita . We suggest that Ins2 Akita mice bone phenotype is caused by lower bone formation and even lower bone resorption due to insulin deficiency and to a possible relation with low leptin signaling.

Comparative landscape genetics reveals the evolution of viviparity reduces genetic connectivity in fire salamanders.

Evolutionary changes in reproductive mode may affect co-evolving traits, such as dispersal, although this subject remains largely underexplored. The shift from aquatic oviparous or larviparous reproduction to terrestrial viviparous reproduction in some amphibians entails skipping the aquatic larval stage and, thus, greater independence from water. Accordingly, amphibians exhibiting terrestrial viviparous reproduction may potentially disperse across a wider variety of suboptimal habitats and increase population connectivity in fragmented landscapes compared to aquatic-breeding species. We investigated this hypothesis in the fire salamander (Salamandra salamandra), which exhibits both aquatic- (larviparity) and terrestrial-breeding (viviparity) strategies. We genotyped 426 larviparous and 360 viviparous adult salamanders for 13 microsatellite loci and sequenced a mitochondrial marker for 133 larviparous and 119 viviparous individuals to compare population connectivity and landscape resistance to gene flow within a landscape genetics framework. Contrary to our predictions, viviparous populations exhibited greater differentiation and reduced genetic connectivity compared to larviparous populations. Landscape genetic analyses indicate viviparity may be partially responsible for this pattern, as water courses comprised a significant barrier only in viviparous salamanders, probably due to their fully terrestrial life cycle. Agricultural areas and, to a lesser extent, topography also decreased genetic connectivity in both larviparous and viviparous populations. This study is one of very few to explicitly demonstrate the evolution of a derived reproductive mode affects patterns of genetic connectivity. Our findings open avenues for future research to better understand the eco-evolutionary implications underlying the emergence of terrestrial reproduction in amphibians.

Resident burnout in USA and European urology residents: an international concern.

OBJECTIVE: To describe the prevalence and predictors of burnout in USA and European urology residents, as although the rate of burnout in urologists is high and associated with severe negative sequelae, the extent and predictors of burnout in urology trainees remains poorly understood. SUBJECTS AND METHODS: An anonymous 32-question survey of urology trainees across the USA and four European countries, analysing personal, programme, and institutional factors, was conducted. Burnout was assessed using the validated abridged Maslach Burnout Inventory. Univariate analysis and multivariable logistic regression models assessed drivers of burnout in the two cohorts. RESULTS: Overall, 40% of participants met the criteria for burnout as follows: Portugal (68%), Italy (49%), USA (38%), Belgium (36%), and France (26%). Response rates were: USA, 20.9%; Italy, 45.2%; Portugal, 30.5%; France, 12.5%; and Belgium, 9.4%. Burnout was not associated with gender or level of training. In both cohorts, work-life balance (WLB) dissatisfaction was associated with increased burnout (odds ratio [OR] 4.5, P < 0.001), whilst non-medical reading (OR 0.6, P = 0.001) and structured mentorship (OR 0.4, P = 0.002) were associated with decreased burnout risk. Lack of access to mental health services was associated with burnout in the USA only (OR 3.5, P = 0.006), whilst more weekends on-call was associated with burnout in Europe only (OR 8.3, P = 0.033). In both cohorts, burned out residents were more likely to not choose a career in urology again (USA 54% vs 19%, P < 0.001; Europe 43% vs 25%, P = 0.047). CONCLUSION: In this study of USA and European urology residents, we found high rates of burnout on both continents. Despite regional differences in the predictors of burnout, awareness of the unique institutional drivers may help inform directions of future interventions.

Accounting for Connectivity Uncertainties in Predicting Roadkills: a Comparative Approach between Path Selection Functions and Habitat Suitability Models.

Functional connectivity modeling is increasingly used to predict the best spatial location for over- or underpasses, to mitigate road barrier effects and wildlife roadkills. This tool requires estimation of resistance surfaces, ideally modeled with movement data, which are costly to obtain. An alternative is to use occurrence data within species distribution models to infer movement resistance, although this remains a controversial issue. This study aimed both to compare the performance of resistance surfaces derived from path versus occurrence data in identifying road-crossing locations of a forest carnivore and assess the influence of movement type (daily vs. dispersal) on this performance. Resistance surfaces were built for genet (Genetta genetta) in southern Portugal using path selection functions with telemetry data, and species distribution models with occurrence data. An independent roadkill dataset was used to evaluate the performance of each connectivity model in predicting roadkill locations. The results show that resistance surfaces derived from occurrence data are as suitable in predicting roadkills as path data for daily movements. When dispersal was simulated, the performance of both resistance surfaces was equally good at predicting roadkills. Moreover, contrary to our expectations, we found no significant differences in locations of roadkill predictions between models based on daily movements and models based on dispersal. Our results suggest that species distribution models are a cost-effective tool to build functional connectivity models for road mitigation plans when movement data are not available.

l-Rhamnosylation of wall teichoic acids promotes efficient surface association of Listeria monocytogenes virulence factors InlB and Ami through interaction with GW domains.

Wall teichoic acids (WTAs) are important surface glycopolymers involved in various physiological processes occurring in the Gram-positive cell envelope. We previously showed that the decoration of Listeria monocytogenes (Lm) WTAs with l-rhamnose conferred resistance against antimicrobial peptides. Here, we show that WTA l-rhamnosylation also contributes to physiological levels of autolysis in Lm through a mechanism that requires efficient association of Ami, a virulence-promoting autolysin belonging to the GW protein family, to the bacterial cell surface. Importantly, WTA l-rhamnosylation also controls the surface association of another GW protein, the invasin internalin B (InlB), that promotes Lm invasion of host cells. Whereas WTA N-acetylglucosaminylation is not a prerequisite for GW protein surface association, lipoteichoic acids appear to also play a role in the surface anchoring of InlB. Strikingly, while the GW domains of Ami, InlB and Auto (another autolysin contributing to cell invasion and virulence) are sufficient to mediate surface association, this is not the case for the GW domains of the remaining six uncharacterized Lm GW proteins. Overall, we reveal WTA l-rhamnosylation as a bacterial surface modification mechanism that contributes to Lm physiology and pathogenesis by controlling the surface association of GW proteins involved in autolysis and infection.