Structure and receptor recognition by the Lassa virus spike complex.

Lassa virus (LASV) is a human pathogen, causing substantial morbidity and mortality1,2. Similar to other Arenaviridae, it presents a class-I spike complex on its surface that facilitates cell entry. The virus's cellular receptor is matriglycan, a linear carbohydrate that is present on α-dystroglycan3,4, but the molecular mechanism that LASV uses to recognize this glycan is unknown. In addition, LASV and other arenaviruses have a unique signal peptide that forms an integral and functionally important part of the mature spike5-8; yet the structure, function and topology of the signal peptide in the membrane remain uncertain9-11. Here we solve the structure of a complete native LASV spike complex, finding that the signal peptide crosses the membrane once and that its amino terminus is located in the extracellular region. Together with a double-sided domain-switching mechanism, the signal peptide helps to stabilize the spike complex in its native conformation. This structure reveals that the LASV spike complex is preloaded with matriglycan, suggesting the mechanism of binding and rationalizing receptor recognition by α-dystroglycan-tropic arenaviruses. This discovery further informs us about the mechanism of viral egress and may facilitate the rational design of novel therapeutics that exploit this binding site.

Involvement of phosphorylated Apis mellifera CREB in gating a honeybee's behavioral response to an external stimulus.

The transcription factor cAMP-response element-binding protein (CREB) is involved in neuronal plasticity. Phosphorylation activates CREB and an increased level of phosphorylated CREB is regarded as an indicator of CREB-dependent transcriptional activation. In honeybees(Apis mellifera)we recently demonstrated a particular high abundance of the phosphorylated honeybee CREB homolog (pAmCREB) in the central brain and in a subpopulation of mushroom body neurons. We hypothesize that these high pAmCREB levels are related to learning and memory formation. Here, we tested this hypothesis by analyzing brain pAmCREB levels in classically conditioned bees and bees experiencing unpaired presentations of conditioned stimulus (CS) and unconditioned stimulus (US). We demonstrate that both behavioral protocols display differences in memory formation but do not alter the level of pAmCREB in bee brains directly after training. Nevertheless, we report that bees responding to the CS during unpaired stimulus presentations exhibit higher levels of pAmCREB than nonresponding bees. In addition, Trichostatin A, a histone deacetylase inhibitor that is thought to enhance histone acetylation by CREB-binding protein, increases the bees' CS responsiveness. We conclude that pAmCREB is involved in gating a bee's behavioral response driven by an external stimulus.

Using the theory of planned behaviour to model antecedents of surgical checklist use: a cross-sectional study.

BACKGROUND: Compliance with surgical checklist use remains an obstacle in the context of checklist implementation programs. The theory of planned behaviour was applied to analyse attitudes, perceived behaviour control, and norms as psychological antecedents of individuals' intentions to use the checklist. METHODS: A cross-sectional survey study with staff (N = 866) of 10 Swiss hospitals was conducted in German and French. Group mean differences between individuals with and without managerial function were computed. Structural equation modelling and confirmatory factor analysis was applied to investigate the structural relation between attitudes, perceived behaviour control, norms, and intentions. RESULTS: Significant mean differences in favour of individuals with managerial function emerged for norms, perceived behavioural control, and intentions, but not for attitudes. Attitudes and perceived behavioural control had a significant direct effect on intentions whereas norms had not. CONCLUSIONS: Individuals with managerial function exhibit stronger perceived behavioural control, stronger norms, and stronger intentions. This could be applied in facilitating checklist implementation. The structural model of the theory of planned behaviour remains stable across groups, indicating a valid model to describe antecedents of intentions in the context of surgical checklist implementation.

Trade-offs between voice and silence: a qualitative exploration of oncology staff's decisions to speak up about safety concerns.

BACKGROUND: Research suggests that "silence", i.e., not voicing safety concerns, is common among health care professionals (HCPs). Speaking up about patient safety is vital to avoid errors reaching the patient and thus to prevent harm and also to improve a culture of teamwork and safety. The aim of our study was to explore factors that affect oncology staff's decision to voice safety concerns or to remain silent and to describe the trade-offs they make. METHODS: In a qualitative interview study with 32 doctors and nurses from 7 oncology units we investigated motivations and barriers to speaking up towards co-workers and supervisors. An inductive thematic content analysis framework was applied to the transcripts. Based on the individual experiences of participants, we conceptualize the choice to voice concerns and the trade-offs involved. RESULTS: Preventing patients from serious harm constitutes a strong motivation to speaking up but competes with anticipated negative outcomes. Decisions whether and how to voice concerns involved complex considerations and trade-offs. Many respondents reflected on whether the level of risk for a patient "justifies" the costs of speaking up. Various barriers for voicing concerns were reported, e.g., damaging relationships. Contextual factors, such as the presence of patients and co-workers in the alarming situation, affect the likelihood of anticipated negative outcomes. Speaking up to well-known co-workers was described as considerably easier whereas "not knowing the actor well" increases risks and potential costs of speaking up. CONCLUSIONS: While doctors and nurses felt strong obligation to prevent errors reaching individual patients, they were not engaged in voicing concerns beyond this immediacy. Our results offer in-depth insight into fears and conditions conducive of silence and voicing and can be used for educational interventions and leader reinforcement.

Safety climate and its association with office type and team involvement in primary care.

OBJECTIVE: To assess differences in safety climate perceptions between occupational groups and types of office organization in primary care. METHODS: Primary care physicians and nurses working in outpatient offices were surveyed about safety climate. Explorative factor analysis was performed to determine the factorial structure. Differences in mean climate scores between staff groups and types of office were tested. Logistic regression analysis was conducted to determine predictors for a 'favorable' safety climate. RESULTS: 630 individuals returned the survey (response rate, 50%). Differences between occupational groups were observed in the means of the 'team-based error prevention'-scale (physician 4.0 vs. nurse 3.8, P < 0.001). Medical centers scored higher compared with single-handed offices and joint practices on the 'team-based error prevention'-scale (4.3 vs. 3.8 vs. 3.9, P < 0.001) but less favorable on the 'rules and risks'-scale (3.5 vs. 3.9 vs. 3.7, P < 0.001). Characteristics on the individual and office level predicted favorable 'team-based error prevention'-scores. Physicians (OR = 0.4, P = 0.01) and less experienced staff (OR 0.52, P = 0.04) were less likely to provide favorable scores. Individuals working at medical centers were more likely to provide positive scores compared with single-handed offices (OR 3.33, P = 0.001). The largest positive effect was associated with at least monthly team meetings (OR 6.2, P < 0.001) and participation in quality circles (OR 4.49, P < 0.001). CONCLUSIONS: Results indicate that frequent quality circle participation and team meetings involving all team members are effective ways to strengthen safety climate in terms of team-based strategies and activities in error prevention.

Average group behavior does not represent individual behavior in classical conditioning of the honeybee.

Conditioned behavior as observed during classical conditioning in a group of identically treated animals provides insights into the physiological process of learning and memory formation. However, several studies in vertebrates found a remarkable difference between the group-average behavioral performance and the behavioral characteristics of individual animals. Here, we analyzed a large number of data (1640 animals) on olfactory conditioning in the honeybee (Apis mellifera). The data acquired during absolute and differential classical conditioning differed with respect to the number of conditioning trials, the conditioned odors, the intertrial intervals, and the time of retention tests. We further investigated data in which animals were tested for spontaneous recovery from extinction. In all data sets we found that the gradually increasing group-average learning curve did not adequately represent the behavior of individual animals. Individual behavior was characterized by a rapid and stable acquisition of the conditioned response (CR), as well as by a rapid and stable cessation of the CR following unrewarded stimuli. In addition, we present and evaluate different model hypotheses on how honeybees form associations during classical conditioning by implementing a gradual learning process on the one hand and an all-or-none learning process on the other hand. In summary, our findings advise that individual behavior should be recognized as a meaningful predictor for the internal state of a honeybee--irrespective of the group-average behavioral performance.

Dissociation of microdissected mouse brain tissue for artifact free single-cell RNA sequencing.

Single-cell RNA sequencing (scRNA-seq) provides the transcriptome of individual cells and addresses previously intractable problems including the central nervous system's transcriptional responses during health and disease. However, dissociating brain cells is challenging and induces artificial transcriptional responses. Here, we describe an enzymatic dissociation method for mouse brain that prevents dissociation artifacts and lowers technical variations with standardized steps. We tested this protocol on microdissected brain tissue of 3-week- to 24-month-old mice and obtained high-quality scRNA-seq results. For complete details on the use and execution of this protocol, please refer to Safaiyan et al. (2021).

Age-associated increase of the active zone protein Bruchpilot within the honeybee mushroom body.

In honeybees, age-associated structural modifications can be observed in the mushroom bodies. Prominent examples are the synaptic complexes (microglomeruli, MG) in the mushroom body calyces, which were shown to alter their size and density with age. It is not known whether the amount of intracellular synaptic proteins in the MG is altered as well. The presynaptic protein Bruchpilot (BRP) is localized at active zones and is involved in regulating the probability of neurotransmitter release in the fruit fly, Drosophila melanogaster. Here, we explored the localization of the honeybee BRP (Apis mellifera BRP, AmBRP) in the bee brain and examined age-related changes in the AmBRP abundance in the central bee brain and in microglomeruli of the mushroom body calyces. We report predominant AmBRP localization near the membrane of presynaptic boutons within the mushroom body MG. The relative amount of AmBRP was increased in the central brain of two-week old bees whereas the amount of Synapsin, another presynaptic protein involved in the regulation of neurotransmitter release, shows an increase during the first two weeks followed by a decrease. In addition, we demonstrate an age-associated modulation of AmBRP located near the membrane of presynaptic boutons within MG located in mushroom body calyces where sensory input is conveyed to mushroom body intrinsic neurons. We discuss that the observed age-associated AmBRP modulation might be related to maturation processes or to homeostatic mechanisms that might help to maintain synaptic functionality in old animals.

Abundance of phosphorylated Apis mellifera CREB in the honeybee's mushroom body inner compact cells varies with age.

Hymenopteran eusociality has been proposed to be associated with the activity of the transcription factor CREB (cAMP-response element binding protein). The honeybee (Apis mellifera) is a eusocial insect displaying a pronounced age-dependent division of labor. In honeybee brains, CREB-dependent genes are regulated in an age-dependent manner, indicating that there might be a role for neuronal honeybee CREB (Apis mellifera CREB, or AmCREB) in the bee's division of labor. In this study, we further explore this hypothesis by asking where in the honeybee brain AmCREB-dependent processes might take place and whether they vary with age in these brain regions. CREB is activated following phosphorylation at a conserved serine residue. An increase of phosphorylated CREB is therefore regarded as an indicator of CREB-dependent transcriptional activation. Thus, we here examine the localization of phosphorylated AmCREB (pAmCREB) in the brain and its age-dependent variability. We report prominent pAmCREB staining in a subpopulation of intrinsic neurons of the mushroom bodies. In these neurons, the inner compact cells (IC), pAmCREB is located in the nuclei, axons, and dendrites. In the central bee brain, the IC somata and their dendritic region, we observed an age-dependent increase of pAmCREB. Our results demonstrate the IC to be candidate neurons involved in age-dependent division of labor. We hypothesize that the IC display a high level of CREB-dependent transcription that might be related to neuronal and behavioral plasticity underlying a bee's foraging behavior.

Effect of a two-year national quality improvement program on surgical checklist implementation.

Use of the surgical checklist in Switzerland is still incomplete and unsatisfactory. A national improvement program was developed and conducted in Switzerland to implement and improve the use of the surgical safety checklists. The aims of the implementation program were to implement comprehensive and correct checklist use in participating hospitals in every patient and in every surgical procedure; and to improve safety climate and teamwork as important cultural context variables. 10 hospitals were selected for participation in the implementation program. A questionnaire assessing use, knowledge, and attitudes towards the checklist and the Safety Climate Survey were conducted at two measurement occasions each in October/November 2013 and January/February 2015. Significant increases emerged for frequency of checklist use (F(1,1001)=340.9, p<0.001), satisfaction (F(1,1232)=25.6, p<0.001), and knowledge(F(1,1294)=184.5, p<0.001). While significant differences in norms (F(1,1284)=17.9, p<0.001) and intentions (F(1,1284)=7.8, p<0.01) were observed, this was not the case for attitudes (F(1,1283)=.8, n.s.) and acceptance (F(1,1284)=0.1, n.s.). Significant differences for safety climate and teamwork emerged in the present study (F(1,3555)=11.8, p<0.001 and F(1,3554)=24.6, p<0.001, respectively). However, although statistical significance was reached, effects are very small and practical relevance is thus questionable. The results of the present study suggest that the quality improvement program conducted by the Swiss Patient Safety Foundation in 10 hospitals led to successful checklist implementation. The strongest effects were seen in aspects concerning behaviour and knowledge specifically related to checklist use. Less impact was achieved on general cultural variables safety climate and teamwork. However, as a trend was observable, these variables may simply need more time in order to change substantially.

Safety climate in Swiss hospital units: Swiss version of the Safety Climate Survey.

RATIONALE, AIMS AND OBJECTIVES: Safety climate measurements are a broadly used element of improvement initiatives. In order to provide a sound and easy-to-administer instrument for the use in Swiss hospitals, we translated the Safety Climate Survey into German and French. METHODS: After translating the Safety Climate Survey into French and German, a cross-sectional survey study was conducted with health care professionals (HCPs) in operating room (OR) teams and on OR-related wards in 10 Swiss hospitals. Validity of the instrument was examined by means of Cronbach's alpha and missing rates of the single items. Item-descriptive statistics group differences and percentage of 'problematic responses' (PPR) were calculated. RESULTS: 3153 HCPs completed the survey (response rate: 63.4%). 1308 individuals were excluded from the analyses because of a profession other than doctor or nurse or invalid answers (n = 1845; nurses = 1321, doctors = 523). Internal consistency of the translated Safety Climate Survey was good (Cronbach's alpha German = 0.86; Cronbach's alpha French = 0.84). Missing rates at item level were rather low (0.23-4.3%). We found significant group differences in safety climate values regarding profession, managerial function, work area and time spent in direct patient care. At item level, 14 out of 21 items showed a PPR higher than 10%. CONCLUSIONS: Results indicate that the French and German translations of the Safety Climate Survey might be a useful measurement instrument for safety climate in Swiss hospital units. Analyses at item level allow for differentiating facets of safety climate into more positive and critical safety climate aspects.

[Patient safety in general practice]. / Patientensicherheit in der Hausarztpraxis.

INTRODUCTION: So far, there has been a lack of systematic data regarding critical incidents and safety climate in Swiss primary care offices. Therefore, a survey was conducted amongst physicians and nurses ("MPA") working in Swiss German primary care offices leading to a subsequent project on the telephone triage. METHODS: Using a standardised questionnaire, healthcare professionals in primary care offices have been surveyed to determine safety risks and safety climate in their offices. The questionnaire consisted of safety-climate items as well as descriptions of 23 safety incidents. These incidents were rated in terms of frequency (appearance in the office during the past 12 months) and severity (harm associated with the last occurrence in the office). In addition, physicians and nurses answered an open-ended question referring to patient safety risks they would wish to eliminate in their offices. In the subsequent project, interviews and group discussions have been conducted with physicians and nurses in order to perform a process analysis of the telephone triage and to develop a tool that may help primary care offices to strengthen telephone triage as a secure process. RESULTS: 630 physicians and nurses (50.2% physicians, 49.8% nurses) participated in the study. 30% of the physicians and 17% of the nurses observed at least one of the 23 incidents in their offices on a daily or weekly basis. Errors in documentation were reported most frequently. As regards severity, the triage by nurses at the initial patient contact, errors in diagnosis, failure to monitor patients after therapeutic treatment in the office, and errors regarding the medication process were shown to be the most relevant. Most frequently participants wanted to eliminate the following risks to patient safety in their offices: medication (28% of all mentions), medical procedures in the office (11%) and telephone triage (7%). Participation in team meetings and quality circles proved to be relevant predictors of the safety climate dimension "team-based error prevention". Differences between occupational groups were found regarding safety incidents as well as safety climate. CONCLUSION: The results of this study show the telephone triage to be a relevant area of patient safety in primary care that has not been focused on so far. In order to enhance safety of the triage process a new project was initiated. The result of the project is a triage guide for primary care offices. This guide supports physicians and nurses in a joint and critical examination of office structures and processes related to telephone triage. The systematically observed differences between occupational groups indicate that the entire office team need to be involved when analysing safety risks and taking action to improve patient safety. Only in doing so, risks can be identified comprehensively. Moreover, measures can be taken that are relevant to and supported by all healthcare professionals working in a primary care office. This approach of involving the entire team forms the basis for the guide on telephone triage.

Silence that can be dangerous: a vignette study to assess healthcare professionals' likelihood of speaking up about safety concerns.

PURPOSE: To investigate the likelihood of speaking up about patient safety in oncology and to clarify the effect of clinical and situational context factors on the likelihood of voicing concerns. PATIENTS AND METHODS: 1013 nurses and doctors in oncology rated four clinical vignettes describing coworkers' errors and rule violations in a self-administered factorial survey (65% response rate). Multiple regression analysis was used to model the likelihood of speaking up as outcome of vignette attributes, responder's evaluations of the situation and personal characteristics. RESULTS: Respondents reported a high likelihood of speaking up about patient safety but the variation between and within types of errors and rule violations was substantial. Staff without managerial function provided significantly higher levels of decision difficulty and discomfort to speak up. Based on the information presented in the vignettes, 74%-96% would speak up towards a supervisor failing to check a prescription, 45%-81% would point a coworker to a missed hand disinfection, 82%-94% would speak up towards nurses who violate a safety rule in medication preparation, and 59%-92% would question a doctor violating a safety rule in lumbar puncture. Several vignette attributes predicted the likelihood of speaking up. Perceived potential harm, anticipated discomfort, and decision difficulty were significant predictors of the likelihood of speaking up. CONCLUSIONS: Clinicians' willingness to speak up about patient safety is considerably affected by contextual factors. Physicians and nurses without managerial function report substantial discomfort with speaking up. Oncology departments should provide staff with clear guidance and trainings on when and how to voice safety concerns.

Communication barriers in counselling foreign-language patients in public pharmacies: threats to patient safety?

BACKGROUND: Foreign-language (FL) patients are at increased risk for adverse drug events. Evidence regarding communication barriers and the safety of pharmaceutical care of FL patients in European countries is scarce despite large migrant populations. OBJECTIVE: To investigate Swiss public pharmacists' experiences and current practices in counselling FL patients with a focus on patient safety. METHOD: In a cross-sectional study heads of public pharmacies in Switzerland were surveyed using an electronic questionnaire. MAIN OUTCOME MEASURE: The survey assessed the frequency of communication barriers encountered in medication counselling of FL patients, perceptions of risks for adverse drug events, satisfaction with the quality of counselling provided to FL patients, current strategies to reduce risks, and preferences towards tools to improve safety for FL patients. RESULTS: 498 pharmacists completed the survey (43 % response rate). More than every second pharmacist reported at least weekly encounters at which they cannot provide good medication counselling to FL patients in the regional Swiss language. Ad-hoc interpreting by minors is also common at a considerable number of pharmacies (26.5 % reported at least one weekly occurrence). Approximately 10 % of pharmacies reported that they fail at least weekly to explain the essentials of drug therapy (e.g. dosing of children's medications) to FL patients. 79.8 % perceived the risk of FL patients for adverse drug events to be somewhat or much higher compared to other patients. 22.5 % of pharmacists reported being concerned at least monthly about medication safety when FL patients leave their pharmacy. However, the majority of pharmacists were satisfied with the quality of care provided to FL patients in their pharmacy [78.6 % (very) satisfied]. The main strategy used to improve counselling for FL patients was the employment of multilingual staff. Participants would use software for printing foreign-language labels (41.2 %) and multilingual package inserts (42.0 %) if these were available. CONCLUSION: Communication barriers with FL patients are frequent in Swiss pharmacies and pharmacists perceive FL patients to be at increased risk for adverse drug events. Development and dissemination of communication tools are needed to support pharmacists in counselling of a diverse migrant population.

Threats to patient safety in the primary care office: concerns of physicians and nurses.

BACKGROUND: Little is known about primary care professionals' concerns about risks to patient safety. AIM: To identify threats to patient safety in the primary care office from the perspective of physicians and nurses. DESIGN: Cross-sectional survey; participants were asked to name and rank threats to safety they personally were most concerned about. SETTING: Physicians and nurses working in primary care offices in Switzerland. METHODS: Verbatim reports were analysed under an inductive content-analysis framework. Coded threats were quantitatively analysed in terms of frequency and prioritisation. Differences between physicians and nurses were analysed. RESULTS: Of 1260 invited individuals, 630 responded to the survey and 391 (31%) described 936 threats to patient safety. The coding system included 29 categories organised in 5 themes. Agreement of coders was good (kappa = 0.87, CI = 0.86-0.87). Safety of medication (8.8%), triage by nurses (7.2%) and drug interactions (6.8%) were the threats cited most frequently. Errors in diagnosis (OR = 0.21, CI 0.09-0.47, p <0.001), drug interactions (OR = 0.10, CI 0.04-0.25, p <0.001) and compliance of patients (OR = 0.28, CI 0.08-0.96, p = 0.044) were more likely to be cited by physicians. X-rays (OR = 3.34, CI 1.04-10.71, p = 0.043), confusion of patients or records (OR = 3.28, CI 1.55-6.94, p = 0.002), hygiene (OR = 3.21, CI1.12-9.19, p = 0.030), safety of office rooms (OR = 6.70, CI 1.46-30.73, p = 0.014), and confidentiality (OR = 7.38, CI 1.63-33.50, p = 0.010) were more likely to be described by nurses. CONCLUSION: Physicians and nurses are concerned about diverse threats to patient safety in primary care. Involving both groups in detection and analysis of risks in medical offices seems a valuable strategy to improve collaboration and safety.

Frequency of and harm associated with primary care safety incidents.

OBJECTIVE: To assess frequency and severity of patient safety incidents in primary care. STUDY DESIGN: Cross-sectional survey of health-care professionals in Swiss primary care offices. METHODS: Physicians and nurses in primary care offices were surveyed about the frequency and severity of 23 safety incidents. Differences between professional groups and types of offices were analyzed. Reported incidents were classified in a matrix. RESULTS: A total of 630 individuals (50.2% physicians, 49.8% nurses) participated. Among them, 30% of physicians (95% confidence interval [CI] 25%-35%) and 16.6% of nurses (95% CI 12%-21%) reported that at least 1 of the incidents occurred daily or weekly in their offices (c2 16.1, P <.001). On average, each responder reported a total of 92 incidents during the preceding 12 months (mean of 117 events for physicians, mean of 66 events for nurses; P <.001). Documentation failure was reported most frequently.The highest fraction of last occurrences with severe injury or death was for diagnostic errors (4.1%). Unadjusted for caseload, staff working in medical centers reported higher frequencies of several incidents. The frequency-harm matrix suggests that triage by nurse at initial contact, diagnostic errors, medication errors, failure to monitor patients after medical procedures, and test or intervention errors should be prioritized for action. CONCLUSIONS: This study presents a supplemental approach to identification of safety threats in primary care. Many incidents occur regularly and are highly relevant for healthcare professionals' daily work.The results offer guidance on setting priorities for patient safety in primary care.

Behavioural pharmacology in classical conditioning of the proboscis extension response in honeybees (Apis mellifera).

Honeybees (Apis mellifera) are well known for their communication and orientation skills and for their impressive learning capability(1,2). Because the survival of a honeybee colony depends on the exploitation of food sources, forager bees learn and memorize variable flower sites as well as their profitability. Forager bees can be easily trained in natural settings where they forage at a feeding site and learn the related signals such as odor or color. Appetitive associative learning can also be studied under controlled conditions in the laboratory by conditioning the proboscis extension response (PER) of individually harnessed honeybees(3,4). This learning paradigm enables the study of the neuronal and molecular mechanisms that underlie learning and memory formation in a simple and highly reliable way(5-12). A behavioral pharmacology approach is used to study molecular mechanisms. Drugs are injected systemically to interfere with the function of specific molecules during or after learning and memory formation(13-16). Here we demonstrate how to train harnessed honeybees in PER conditioning and how to apply drugs systemically by injection into the bee flight muscle.