Diversifying selection identified in immune epitopes of bovine coronavirus isolates from Irish cattle.

Bovine betacoronavirus (BoCoV) is a pneumoenteric pathogen of cattle that is closely related to human coronavirus OC43. Vaccines are administered to protect against diseases caused by BoCoV, but knowledge gaps exist with regard to correlates of protection and the effect of immune evasion on driving evolution. In this study, immune epitopes were mapped onto BoCoV structural proteins, including spike and haemagglutinin esterase (HE), and then supported with targeted gene sequencing of Irish clinical isolates and selective pressure analysis. Increased prevalence of diversifying selection and amino acid changes in some mapped immune epitopes suggests that immune escape is selecting for non-synonymous mutations arising in these regions. Selection analysis and sequencing provided increased support for neutralising antibody (nAb) epitopes compared to others, suggesting that nAbs are an important arm of the immune response to BoCoV. Phylogenetic analysis of spike and HE sequences showed that Irish isolates from this study were in the European clade, except for one HE sequence that sat in the Asian/American clade, while the spike gene of this sample was in the European clade. Recombination between a European and an Asian/American isolate would give rise to such a sequence. This study has gathered evidence suggesting that pressure to evade the nAb response is contributing to BoCoV evolution.

Effect of COVID-19 on Injury Incidence and Severity in Professional Female Football Players: A Cohort Prospective Study.

BACKGROUND: Earlier statements suggested a negative impact of coronavirus disease 2019 (COVID-19) infection on sports performance and injury risk. With the COVID-19 pandemic under control and the dominance of a less-severe strain of the virus, there is a need to confirm whether these adverse effects still apply to the current situation. HYPOTHESIS: Infected players would have a higher noncontact muscle injury incidence compared with noninfected counterparts. STUDY DESIGN: Cohort observational study. LEVEL OF EVIDENCE: Level 3. METHODS: Seven teams (n = 147 players) competing in the Spanish professional women's football league (Liga F) were prospectively monitored during the 2021-2022 season. Data from noncontact injuries were recorded and classified following the latest consensus statement from the International Olympic Committee. COVID-19 was certified by the medical staff by regular polymerase chain reaction analysis. RESULTS: Ninety-two players suffered at least 1 noncontact muscle injury during the season. Injury incidence during the season was similar in players with COVID-19 (n = 83) and players without infection (5.1 ± 6.7 versus 4.9 ± 10.0 injuries/1000 h of play, respectively; P = 0.90). Players with COVID-19 were not more likely to suffer noncontact injuries compared with those players without infection (R2 = 0.02; odds ratio [OR] 95% confidence interval [95% CI] = 0.36-1.38; P = 0.31). There was no effect of COVID-19 on the days of absence due to injury (R2 = 0.01; OR 95% CI = 1.00-1.01; P = 0.44) or in the classification of the severity of the injury (P = 0.79). CONCLUSION: COVID-19 has no significant effect on noncontact injury incidence and severity in professional female football players. CLINICAL RELEVANCE: Currently, COVID-19 infection does not alter noncontact muscle injury risk in professional football and requires no further attention in terms of injury management. Usual return-to-play protocols apply to COVID-19 considering the particularities of each player since the severity of infection, period of inactivity, and effects on the player's health and performance.

Amphetamine disrupts dopamine axon growth in adolescence by a sex-specific mechanism in mice.

Initiating drug use during adolescence increases the risk of developing addiction or other psychopathologies later in life, with long-term outcomes varying according to sex and exact timing of use. The cellular and molecular underpinnings explaining this differential sensitivity to detrimental drug effects remain unexplained. The Netrin-1/DCC guidance cue system segregates cortical and limbic dopamine pathways in adolescence. Here we show that amphetamine, by dysregulating Netrin-1/DCC signaling, triggers ectopic growth of mesolimbic dopamine axons to the prefrontal cortex, only in early-adolescent male mice, underlying a male-specific vulnerability to enduring cognitive deficits. In adolescent females, compensatory changes in Netrin-1 protect against the deleterious consequences of amphetamine on dopamine connectivity and cognitive outcomes. Netrin-1/DCC signaling functions as a molecular switch which can be differentially regulated by the same drug experience as function of an individual's sex and adolescent age, and lead to divergent long-term outcomes associated with vulnerable or resilient phenotypes.

Effect of playing position and microcycle days on the acceleration speed profile of elite football players.

The aim of this study was to analyse the differences in the A-S profile of elite football players induced by playing position and the microcycle day. Players belonged to a second division club in the Spanish La Liga competition. They were classified into five playing positions: central defenders (CD), full backs (FB), midfielders (MF), wide midfielders (WMF) and forwards (FW). Microcycle days were categorised according to the days until matchday (MD, MD-1, MD-2, MD-3, MD-4 and MD-5). Data was collected along six microcycles, including one match per microcycle. The variables analysed were: maximal theoretical acceleration (A0), maximal theoretical speed (S0), maximal acceleration (ACCmax), maximal speed (Smax) and A-S slope (ASslope). Significant differences were found within positions and microcycle day for all variables (p < 0.05). Match day (MD) showed greater values than the training sessions in A0, ACCmax and Smax (p < 0.05). The highest values for variables associated with acceleration capabilities were found in CD on MD, whereas speed variables were higher in WMF. MD-2 showed the lowest values in all variables except for ASslope. Maximal acceleration and sprint abilities are therefore affected by playing position. Wide positions showed the highest speed capacity, and CD presented a likely acceleration profile. Higher values for all variables concerning the microcycle day, were achieved on MD, and were not reproduced during training with the consequent injury risk and performance decrease it takes.

Spain is not different: teaching quantitative courses can also be hazardous to one's career (at least in undergraduate courses).

Student evaluations of teaching (SETs) have become a widely used tool for assessing teaching in higher education. However, numerous investigations have shown that SETs are subject to multiple biases, one of which is particularly relevant, namely, the area of knowledge to which the subject belongs. This article aims to replicate the article by Uttl & Smibert (2017, https://doi.org/10.7717/peerj.3299) in a different educational context to verify whether the negative bias toward instructors who teach quantitative courses found by the authors in the US also appears in the Spanish university system. The study was conducted at the Business and Law School of the Universidad Pontificia Comillas, a private Spanish university, using two different samples. First, we analyzed undergraduate courses using a sample of 80,667 SETs in which 2,885 classes (defined as a single semester-long course taught by an individual instructor to a specific group of students), 488 instructors, and 322 different courses were evaluated over a time period of four academic years (2016/2017-2019/2020). Second, in the same period, 16,083 SETs corresponding to master's degree courses were analyzed, which involved the study of 871 classes, 275 instructors, and 155 different courses. All the data included in the analysis were obtained from official university surveys developed by a team of professionals specialized in teaching quality responsible for ensuring the reliability of the information. At the degree level, the results show that despite the considerable cultural and temporal difference between the samples, the results are very similar to those obtained by Uttl & Smibert (2017, https://doi.org/10.7717/peerj.3299); i.e., professors teaching quantitative courses are far more likely to obtain worse SETs than instructors in other areas. There are hardly any differences at the master's degree level, regardless of whether nearly 75% of master's degree instructors also teach at the undergraduate level. This leads us to three different conclusions. (1) Evidence suggests that the reason for these differences is not due to faculty teaching quantitative courses being less effective than faculty teaching in some other fields. Our results indicate that the same instructor is evaluated very differently depending on whether he or she teaches at the undergraduate or master's level. (2) It is essential to avoid comparisons of SETs between different areas of knowledge, at least at the undergraduate level. (3) A significant change in the use and interpretation of SETs is imperative, or its replacement by other evaluation mechanisms should be considered. If this does not occur, it is possible that in the future, there will be an adverse selection effect among professors of quantitative methods; i.e., only the worst professionals in quantitative methods will opt for teaching since the good professionals will prefer other jobs.

Identifying Targets for Antibiotic Use for the Management of Carbapenem-Resistant Acinetobacter baumannii (CRAb) in Hospitals-A Multi-Centre Nonlinear Time-Series Study.

Solutions are needed to inform antimicrobial stewardship (AMS) regarding balancing the access to effective antimicrobials with the need to control antimicrobial resistance. Theoretical and mathematical models suggest a non-linear relationship between antibiotic use and resistance, indicating the existence of thresholds of antibiotic use beyond which resistance would be triggered. It is anticipated that thresholds may vary across populations depending on host, environment, and organism factors. Further research is needed to evaluate thresholds in antibiotic use for a specific pathogen across different settings. The objective of this study is to identify thresholds of population antibiotic use associated with the incidence of carbapenem-resistant Acinetobacter baumannii (CRAb) across six hospital sites in Oman. The study was an ecological, multi-centre evaluation that involved collecting historical antibiotic use and CRAb incidence over the period from January 2015 to December 2019. By using non-linear time-series analysis, we identified different thresholds in the use of third-generation cephalosporins, piperacillin-tazobactam, aminoglycoside, and fluoroquinolones across participating hospitals. The identification of different thresholds emphasises the need for tailored analysis based on modelling data from each hospital. The determined thresholds can be used to set targets for each hospital AMS, providing a balance between access to these antibiotics versus controlling CRAb incidence.

Long-Term Impact of the COVID-19 Pandemic on In-Hospital Antibiotic Consumption and Antibiotic Resistance: A Time Series Analysis (2015-2021).

The coronavirus disease 2019 (COVID-19)-pandemic-related overload of health systems has compromised the application of antimicrobial stewardship (AS) models and infection prevention and control (IPC) programs. We aimed to evaluate the impact of COVID-19 on antimicrobial consumption (AC) and antimicrobial resistance (AMR) in the University Hospital of Modena. A time series analysis with an autoregressive integrated moving average model was conducted from January 2015 to October 2021 to evaluate the AC in the whole hospital and the intensive care unit (ICU), the incidence density (ID) of bloodstream infections (BSIs) due to the main multidrug-resistant organisms, and of C. difficile infections (CDIs). After an initial peak during the COVID-19 period, a decrease in the trend of AC was observed, both at the hospital (CT: -1.104, p = 0.025) and ICU levels (CT: -4.47, p = 0.047), with no significant difference in the single classes. Among the Gram-negative isolates, we observed a significant increase only in the level of BSIs due to carbapenem-susceptible Pseudomonas aeruginosa (CL: 1.477, 95% CI 0.130 to 2.824, p = 0.032). Considering Gram-positive bacteria, an increase in the level of BSIs due to methicillin-resistant Staphylococcus aureus and in the trend of CDIs were observed, though they did not reach statistical significance (CL: 0.72, 95% CI -0.039 to 1.48, p = 0.062; CT: 1.43, 95% CI -0.002 to 2.863, p = 0.051; respectively). Our findings demonstrated that the increases in AMR and AC that appeared in the first COVID-19 wave may be later controlled by restoring IPC and AS programs to pre-epidemic levels. A coordinated healthcare effort is necessary to address the longer-term impact of COVID-19 on AC to avoid irreversible consequences on AMR.

A Herd Investigation Tool in Support of the Irish Bovine Viral Diarrhoea Eradication Programme.

Bovine viral diarrhoea (BVD) is an important endemic disease of cattle. In Ireland, an industry-led compulsory eradication programme began in January 2013. The main elements of this programme are the identification and elimination of persistently infected (PI) calves by testing all new-borns, the implementation of biosecurity to prevent re-introduction of disease and continuous surveillance. In 2016, a standardised framework was developed to investigate herds with positive results. This is delivered by trained private veterinary practitioners (PVP). The investigation's aims are 3-fold: firstly, to identify plausible sources of infection; secondly, to ensure that no virus-positive animals remain on farm by resolving the BVD status of all animals in the herd; and thirdly, agreeing up to three biosecurity measures with the herd owner to prevent the re-introduction of the virus. Each investigation follows a common approach comprising four steps based on information from the programme database and collected on-farm: firstly, identifying the time period when each virus-positive calf was exposed in utero (window of susceptibility, taken as 30-120 days of gestation); secondly, determining the location of the dam of each positive calf during this period; thirdly, to investigate potential sources of exposure, either within the herd or external to it; and finally, based on the findings, the PVP and herdowner agree to implement up to three biosecurity measures to minimise the risk of reintroduction. Between 2016 and 2020, 4,105 investigations were completed. The biosecurity recommendations issued more frequently related to the risks of introduction of virus associated with contact with neighbouring cattle at pasture, personnel (including the farmer), the purchase of cattle and vaccination. Although each investigation generates farm-specific outcomes and advice, the aggregated results also provide an insight into the most commonly identified transmission pathways for these herds which inform overall programme communications on biosecurity. The most widely identified plausible sources of infection over these years included retained BVD-positive animals, Trojan births, contact at boundaries and indirect contact through herd owner and other personnel in the absence of appropriate hygiene measures. While generated in the context of BVD herd investigations, the findings also provide an insight into biosecurity practises more generally on Irish farms.

First detected case of rabbit Haemorrhagic disease virus 2 (RHDV2) in the Irish hare (Lepus timidus hibernicus).

BACKGROUND: Rabbit haemorrhagic disease virus (RHDV) is a Lagovirus, a subgroup of the family Caliciviridae. RHDV2 is a variant first described in France in 2010, and has since spread globally. It has been reported in several Lagomorph species (rabbits, hares, and their relatives) as well as other mammals including voles and shrews. The disease has raised international concerns for its potential impact on population abundance trajectories, particularly as 25% of Lagomorphs are currently Red-Listed by the International Union for the Conservation of Nature (IUCN). The Irish hare (Lepus timidus hibernicus) is a subspecies of the mountain hare, L. timidus, and is endemic to Ireland, making it an Evolutionarily Significant Unit of intrinsic value. CASE PRESENTATION: The first case of RHDV2 was detected in a wild Irish hare in July 2019. The individual exhibited atypical neurological behaviour (running in circles) prior to death. On necropsy, pink tinged foam was seen in the trachea and congestion was noted in the lungs, but there was no evidence of haemorrhages in any other organ. Both the liver and spleen were tested by reverse transcription real time qPCR confirming high levels of RHDV2 RNA. Histopathology confirmed multifocal necrotising hepatitis. CONCLUSION: The Irish hare is susceptible to RHDV2 infection. Further investigation is warranted to explore the clinical, epidemiological, and population biology implications.

A five-component infection control bundle to permanently eliminate a carbapenem-resistant Acinetobacter baumannii spreading in an intensive care unit.

BACKGROUND: Carbapenem-resistant Acinetobacter baumannii (CRAB) infection outbreaks are difficult to control and sometimes require cohorting of CRAB-positive patients or temporary ward closure for environmental cleaning. We aimed at controlling the deadly 2018 CRAB outbreak in a 12 bed- intensive care unit (ICU) including 9 beds in a 220 m2 open space. We implemented a new multimodal approach without ward closure, cohorting or temporarily limiting admissions. METHODS: A five-component bundle was introduced in 2018 including reinforcement of hand hygiene and sample extension of screening, application of contact precautions to all patients, enhanced environmental sampling and the one-time application of a cycling radical environmental cleaning and disinfection procedure of the entire ICU. The ICU-CRAB incidence density (ID), ICU alcohol-based hand rub consumption and antibiotic use were calculated over a period of 6 years and intervention time series analysis was performed. Whole genome sequencing analysis (WGS) was done on clinical and environmental isolates in the study period. RESULTS: From January 2013, nosocomial ICU-CRAB ID decreased from 30.4 CRAB cases per 1000 patients-days to zero cases per 1000 patients-days. Our intervention showed a significant impact (-2.9 nosocomial ICU-CRAB cases per 1000 bed-days), while no influence was observed for antibiotic and alcohol-based hand rub (AHR) consumption. WGS demonstrated that CRAB strains were clonally related to an environmental reservoir which confirms the primary role of the environment in CRAB ICU spreading. CONCLUSION: A five-component bundle of continuous hand hygiene improvement, extended sampling at screening including the environment, universal contact precautions and a novel cycling radical environmental cleaning and disinfection procedure proved to be effective for permanently eliminating CRAB spreading within the ICU. Cohorting, admission restriction or ICU closure were avoided.

The Irish Programme to Eradicate Bovine Viral Diarrhoea Virus-Organization, Challenges, and Progress.

A mandatory national Irish bovine viral diarrhoea (BVD) eradication programme, coordinated by Animal Health Ireland, commenced in 2013. Key decisions and programme review are undertaken by a cross-industry Implementation Group (BVDIG) supported by a Technical Working Group. Ear notch tissue is collected from all new-born calves using modified official identity tags, supplemented by additional blood sampling, including for confirmatory testing of calves with initial positive results and testing of their dams. Testing is delivered by private laboratories in conjunction with the National Reference Laboratory, with all results reported to a central database. This database manages key elements of the programme, issuing results to herdowners by short message service messaging supplemented by letters; assigning and exchanging animal-level statuses with government databases of the Department of Agriculture, Food and the Marine to enable legislated restrictions on animal movements; assigning negative herd status based on test results; generating regular reports for programme management and evaluation and providing herd-specific dashboards for a range of users. Legislation supporting the programme has been in place throughout but has not thus far mandated the slaughter of persistently infected (PI) calves. A key challenge in the early years, highlighted by modeling, was the retention of PI animals by some herd owners. This has largely been resolved by measures including graduated financial supports to encourage their early removal, herd-level movement restrictions, ongoing programme communications and the input of private veterinary practitioners (PVPs). A framework for funded investigations by PVPs in positive herds was developed to identify plausible sources of infection, to resolve the status of all animals in the herd and to agree up to three measures to prevent re-introduction of the virus. The prevalence of PI calves in 2013 was 0.66%, within 11.3% of herds, reducing in each subsequent year, to 0.03 and 0.55%, respectively, at the end of 2020. Recent regulatory changes within the European Union for the first time make provision for official approval of national eradication programmes, or recognition of BVD freedom, and planning is underway to seek approval and, in due course, recognition of freedom within this framework by 2023.

Identification of thresholds in relationships between specific antibiotic use and carbapenem-resistant Acinetobacter baumannii (CRAb) incidence rates in hospitalized patients in Jordan.

BACKGROUND: Antibiotic resistance is a major threat to public health worldwide. The relationship between the intensity of antibiotic use and resistance might not be linear, suggesting that there might be a threshold of antibiotic use, beyond which resistance would be triggered. OBJECTIVES: To identify thresholds in antibiotic use, below which specific antibiotic classes have no significant measurable impact on the incidence of carbapenem-resistant Acinetobacter baumannii (CRAb), but above which their use correlates with an increase in the incidence of CRAb. METHODS: The study took place at a tertiary teaching hospital in Jordan. The study was ecological in nature and was carried out retrospectively over the period January 2014 to December 2019. The outcome time series for this study was CRAb cases. The primary explanatory variables were monthly use of antibiotics and the use of alcohol-based hand rub (ABHR). Non-linear time-series methods were used to identify thresholds in antibiotic use. RESULTS: Non-linear time-series analysis determined a threshold in third-generation cephalosporin and carbapenem use, where the maximum use of third-generation cephalosporins and carbapenems should not exceed 8 DDD/100 occupied bed days (OBD) and 10 DDD/100 OBD, respectively. ABHR had a significant reducing effect on CRAb cases even at lower usage quantities (0.92 L/100 OBD) and had the most significant effect when ABHR exceeded 3.4 L/100 OBD. CONCLUSIONS: The identification of thresholds, utilizing non-linear time-series methods, can provide a valuable tool to inform hospital antibiotic policies through identifying quantitative targets that balance access to effective therapies with control of resistance. Further studies are needed to validate the identified thresholds, through being prospectively adopted as a target for antimicrobial stewardship programmes, and then to evaluate the impact on reducing CRAb incidence.

Match and Training High Intensity Activity-Demands Profile During a Competitive Mesocycle in Youth Elite Soccer Players.

The monitoring of the high intensity activity-demands profile during official matches (OMs) and training sessions (TSs) provides a deeper understanding of the relationship between training and competition loads as well as players' fitness characteristics. The aims of this study were to: 1) describe the training and match high intensity activity-demands profile in U-19 soccer players; 2) compare the profile depending on the type of session (OM or TS) throughout match-weeks; and 3) differentiate between profiles depending on the match location (home or away). Twenty-five U-19 Spanish soccer players were monitored during TSs and OMs for a one-month competitive period using a WIMU PROTM wearable inertial device. The variables of the study were: high speed running distance (HSRD), total sprints (SPs), maximum speed (MS) and player load (PL). OMs required higher demands than TSs in HSRD (460.99 ± 206.18 vs. 315.45 ± 180.12 m; p < 0.01; d = 0.75), SPs (10.86 ± 6.64 vs. 7.23 ± 4.82; p < 0.01; d = 0.69), MS (29.99 ± 2.54 vs. 28.50 ± 2.4 km/h; p < 0.01; d = 0.59) and PL (103.08 ± 24.15 vs. 83.18 ± 17.96 a.u.; p < 0.01; d = 0.94). The interaction between the type of session and mean week's demands presented differences with medium effect size in MS (p < 0.01; ωp 2 = 0.06) and small effect size in HSRD (p = 0.04; ωp 2 = 0.03), and SP (p = 0.05; ωp 2 = 0.03), but there were no differences in PL (p = 0.18; ωp 2 = 0). Finally, no differences were found in the match location comparison (p > 0.33; d = 0.22-0.33). Therefore, the profiles presented could be useful for future scientific purposes and serve as valid information for coaches trying to optimize performance.

Author Correction: A nonlinear time-series analysis approach to identify thresholds in associations between population antibiotic use and rates of resistance.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

DCC-related developmental effects of abused- versus therapeutic-like amphetamine doses in adolescence.

The guidance cue receptor DCC controls mesocortical dopamine development in adolescence. Repeated exposure to an amphetamine regimen of 4 mg/kg during early adolescence induces, in male mice, downregulation of DCC expression in dopamine neurons by recruiting the Dcc microRNA repressor, microRNA-218 (miR-218). This adolescent amphetamine regimen also disrupts mesocortical dopamine connectivity and behavioral control in adulthood. Whether low doses of amphetamine in adolescence induce similar molecular and developmental effects needs to be established. Here, we quantified plasma amphetamine concentrations in early adolescent mice following a 4 or 0.5 mg/kg dose and found peak levels corresponding to those seen in humans following recreational and therapeutic settings, respectively. In contrast to the high doses, the low amphetamine regimen does not alter Dcc mRNA or miR-218 expression; instead, it upregulates DCC protein levels. Furthermore, high, but not low, drug doses downregulate the expression of the DCC receptor ligand, Netrin-1, in the nucleus accumbens and prefrontal cortex. Exposure to the low-dose regimen did not alter the expanse of mesocortical dopamine axons or their number/density of presynaptic sites in adulthood. Strikingly, adolescent exposure to the low-dose drug regimen does not impair behavioral inhibition in adulthood; instead, it induces an overall increase in performance in a go/no-go task. These results show that developmental consequences of exposure to therapeutic- versus abused-like doses of amphetamine in adolescence have dissimilar molecular signatures and opposite behavioral effects. These findings have important clinical relevance since amphetamines are widely used for therapeutic purposes in youth.

MiR-218: a molecular switch and potential biomarker of susceptibility to stress.

Low miR-218 expression in the medial prefrontal cortex (mPFC) is a consistent trait of depression. Here we assessed whether miR-218 in the mPFC confers resilience or susceptibility to depression-like behaviors in adult mice, using the chronic social defeat stress (CSDS) model of depression. We also investigated whether stress-induced variations of miR-218 expression in the mPFC can be detected in blood. We find that downregulation of miR-218 in the mPFC increases susceptibility to a single session of social defeat, whereas overexpression of miR-218 selectively in mPFC pyramidal neurons promotes resilience to CSDS and prevents stress-induced morphological alterations to those neurons. After CSDS, susceptible mice have low levels of miR-218 in blood, as compared with control or resilient groups. We show further that upregulation and downregulation of miR-218 levels specifically in the mPFC correlate with miR-218 expression in blood. Our results suggest that miR-218 in the adult mPFC might function as a molecular switch that determines susceptibility vs. resilience to chronic stress, and that stress-induced variations in mPFC levels of miR-218 could be detected in blood. We propose that blood expression of miR-218 might serve as potential readout of vulnerability to stress and as a proxy of mPFC function.

A nonlinear time-series analysis approach to identify thresholds in associations between population antibiotic use and rates of resistance.

Balancing access to antibiotics with the control of antibiotic resistance is a global public health priority. At present, antibiotic stewardship is informed by a 'use it and lose it' principle, in which antibiotic use by the population is linearly related to resistance rates. However, theoretical and mathematical models suggest that use-resistance relationships are nonlinear. One explanation for this is that resistance genes are commonly associated with 'fitness costs' that impair the replication or transmissibility of the pathogen. Therefore, resistant genes and pathogens may only gain a survival advantage where antibiotic selection pressures exceed critical thresholds. These thresholds may provide quantitative targets for stewardship-optimizing the control of resistance while avoiding over-restriction of antibiotics. Here, we evaluated the generalizability of a nonlinear time-series analysis approach for identifying thresholds using historical prescribing and microbiological data from five populations in Europe. We identified minimum thresholds in temporal relationships between the use of selected antibiotics and incidence rates of carbapenem-resistant Acinetobacter baumannii (Hungary), extended-spectrum ß-lactamase-producing Escherichia coli (Spain), cefepime-resistant E. coli (Spain), gentamicin-resistant Pseudomonas aeruginosa (France) and methicillin-resistant Staphylococcus aureus (Northern Ireland) in different epidemiological phases. Using routinely generated data, our approach can identify context-specific quantitative targets for rationalizing population antibiotic use and controlling resistance. Prospective intervention studies that restrict antibiotic consumption are needed to validate these thresholds.

Dopamine Development in the Mouse Orbital Prefrontal Cortex Is Protracted and Sensitive to Amphetamine in Adolescence.

The prefrontal cortex (PFC) is divided into subregions, including the medial and orbital prefrontal cortices. Dopamine connectivity in the medial PFC (mPFC) continues to be established throughout adolescence as the result of the continuous growth of axons that innervated the nucleus accumbens (NAcc) prior to adolescence. During this period, dopamine axons remain vulnerable to environmental influences, such as drugs used recreationally by humans. The developmental trajectory of the orbital prefrontal dopamine innervation remains almost completely unstudied. Nonetheless, the orbital PFC (oPFC) is critical for some of the most complex functions of the PFC and is disrupted by drugs of abuse, both in adolescent humans and rodents. Here, we use quantitative neuroanatomy, axon-initiated viral-vector recombination, and pharmacology in mice to determine the spatiotemporal development of the dopamine innervation to the oPFC and its vulnerability to amphetamine in adolescence. We find that dopamine innervation to the oPFC also continues to increase during adolescence and that this increase is due to the growth of new dopamine axons to this region. Furthermore, amphetamine in adolescence dramatically reduces the number of presynaptic sites on oPFC dopamine axons. In contrast, dopamine innervation to the piriform cortex is not protracted across adolescence and is not impacted by amphetamine exposure during adolescence, indicating that dopamine development during adolescence is a uniquely prefrontal phenomenon. This renders these fibers, and the PFC in general, particularly vulnerable to environmental risk factors during adolescence, such as recreational drug use.

Non-Contingent Exposure to Amphetamine in Adolescence Recruits miR-218 to Regulate Dcc Expression in the VTA.

The development of the dopamine input to the medial prefrontal cortex occurs during adolescence and is a process that is vulnerable to disruption by stimulant drugs such as amphetamine. We have previously linked the amphetamine-induced disruption of dopamine connectivity and prefrontal cortex maturation during adolescence to the downregulation of the Netrin-1 receptor, DCC, in dopamine neurons. However, how DCC expression in dopamine neurons is itself regulated is completely unknown. MicroRNA (miRNA) regulation of mRNA translation and stability is a prominent mechanism linking environmental events to changes in protein expression. Here, using male mice, we show that miR-218 is expressed in dopamine neurons and is a repressor of DCC. Whereas Dcc mRNA levels increase from early adolescence to adulthood, miR-218 exhibits the exact opposite switch, most likely maintaining postnatal Dcc expression. This dynamic regulation appears to be selective to Dcc since the expression of Robo 1, the other guidance cue receptor target of miR-218, does not vary with age. Amphetamine in adolescence, but not in adulthood, increases miR-218 in the VTA and this event is required for drug-induced downregulation of Dcc mRNA and protein expression. This effect seems to be specific to Dcc because amphetamine does not alter Robo1. Furthermore, the upregulation of miR-218 by amphetamine requires dopamine D2 receptor activation. These findings identify miR-218 as regulator of DCC in the VTA both in normal development and after drug exposure in adolescence.