Dialyzing a Brain-Dead Individual for Organ Procurement.

Many patients are unable to receive organ transplantation as there is an expanding gap between the number of patients waiting for an organ and the number who receive it. Organ procurement from the brain-dead can address this expanding gap, especially because one brain-dead patient can potentially donate multiple organs to several recipients. Here, we describe a rare case of a previously healthy 26-year-old male who was declared brain dead after a motor vehicle accident but underwent hemodialysis to treat his acute kidney injury and hyperkalemia before successfully donating his heart and left kidney.

Aflatoxin B1 Increases Soluble Epoxide Hydrolase in the Brain and Induces Neuroinflammation and Dopaminergic Neurotoxicity.

Parkinson's disease (PD) is an increasingly common neurodegenerative movement disorder with contributing factors that are still largely unexplored and currently no effective intervention strategy. Epidemiological and pre-clinical studies support the close association between environmental toxicant exposure and PD incidence. Aflatoxin B1 (AFB1), a hazardous mycotoxin commonly present in food and environment, is alarmingly high in many areas of the world. Previous evidence suggests that chronic exposure to AFB1 leads to neurological disorders as well as cancer. However, whether and how aflatoxin B1 contributes to the pathogenesis of PD is poorly understood. Here, oral exposure to AFB1 is shown to induce neuroinflammation, trigger the α-synuclein pathology, and cause dopaminergic neurotoxicity. This was accompanied by the increased expression and enzymatic activity of soluble epoxide hydrolase (sEH) in the mouse brain. Importantly, genetic deletion or pharmacological inhibition of sEH alleviated the AFB1-induced neuroinflammation by reducing microglia activation and suppressing pro-inflammatory factors in the brain. Furthermore, blocking the action of sEH attenuated dopaminergic neuron dysfunction caused by AFB1 in vivo and in vitro. Together, our findings suggest a contributing role of AFB1 to PD etiology and highlight sEH as a potential pharmacological target for alleviating PD-related neuronal disorders caused by AFB1 exposure.

Immunocytoprotection after reperfusion with Kv1.3 inhibitors has an extended treatment window for ischemic stroke.

Introduction: Mechanical thrombectomy has improved treatment options and outcomes for acute ischemic stroke with large artery occlusion. However, as the time window of endovascular thrombectomy is extended there is an increasing need to develop immunocytoprotective therapies that can reduce inflammation in the penumbra and prevent reperfusion injury. We previously demonstrated, that by reducing neuroinflammation, KV1.3 inhibitors can improve outcomes not only in young male rodents but also in female and aged animals. To further explore the therapeutic potential of KV1.3 inhibitors for stroke therapy, we here directly compared a peptidic and a small molecule KV1.3 blocker and asked whether KV1.3 inhibition would still be beneficial when started at 72 hours after reperfusion. Methods: Transient middle cerebral artery occlusion (tMCAO, 90-min) was induced in male Wistar rats and neurological deficit assessed daily. On day-8 infarction was determined by T2-weighted MRI and inflammatory marker expression in the brain by quantitative PCR. Potential interactions with tissue plasminogen activator (tPA) were evaluated in-vitro with a chromogenic assay. Results: In a direct comparison with administration started at 2 hours after reperfusion, the small molecule PAP-1 significantly improved outcomes on day-8, while the peptide ShK-223 failed to reduce infarction and neurological deficits despite reducing inflammatory marker expression. PAP-1 still provided benefits when started 72 hours after reperfusion. PAP-1 does not reduce the proteolytic activity of tPA. Discussion: Our studies suggest that KV1.3 inhibition for immunocytoprotection after ischemic stroke has a wide therapeutic window for salvaging the inflammatory penumbra and requires brain-penetrant small molecules.

Repurposing the KCa3.1 Blocker Senicapoc for Ischemic Stroke.

Senicapoc, a small molecule inhibitor of the calcium-activated potassium channel KCa3.1, was safe and well-tolerated in clinical trials for sickle cell anemia. We previously reported proof-of-concept data suggesting that both pharmacological inhibition and genetic deletion of KCa3.1 reduces infarction and improves neurologic recovery in rodents by attenuating neuroinflammation. Here we evaluated the potential of repurposing senicapoc for ischemic stroke. In cultured microglia, senicapoc inhibited KCa3.1 currents with an IC50 of 7 nM, reduced Ca2+ signaling induced by the purinergic agonist ATP, suppressed expression of pro-inflammatory cytokines and enzymes (iNOS and COX-2), and prevented induction of the inflammasome component NLRP3. When transient middle cerebral artery occlusion (tMCAO, 60 min) was induced in male C57BL/6 J mice, twice daily administration of senicapoc at 10 and 40 mg/kg starting 12 h after reperfusion dose-dependently reduced infarct area determined by T2-weighted magnetic resonance imaging (MRI) and improved neurological deficit on day 8. Ultra-high-performance liquid chromatography/mass spectrometry analysis of total and free brain concentrations demonstrated sufficient KCa3.1 target engagement. Senicapoc treatment significantly reduced microglia/macrophage and T cell infiltration and activation and attenuated neuronal death. A different treatment paradigm with senicapoc started at 3 h and MRI on day 3 and day 8 revealed that senicapoc reduces secondary infarct growth and suppresses expression of inflammation markers, including T cell cytokines in the brain. Lastly, we demonstrated that senicapoc does not impair the proteolytic activity of tissue plasminogen activator (tPA) in vitro. We suggest that senicapoc could be repurposed as an adjunctive immunocytoprotective agent for combination with reperfusion therapy for ischemic stroke.

Contextual modulation of appearance-trait learning.

When we encounter a stranger for the first time, we spontaneously attribute to them a wide variety of character traits based on their facial appearance. There is increasing consensus that learning plays a key role in these first impressions. According to the Trait Inference Mapping (TIM) model, first impressions are the products of mappings between 'face space' and 'trait space' acquired through domain-general associative processes. Drawing on the associative learning literature, TIM predicts that first-learned associations between facial appearance and character will be particularly influential: they will be difficult to unlearn and will be more likely to generalise to novel contexts than appearance-trait associations acquired subsequently. The study of face-trait learning de novo is complicated by the fact that participants, even young children, already have extensive experience with faces before they enter the lab. This renders the study of first-learned associations from faces intractable. Here, we overcome this problem by using Greebles - a class of novel synthetic objects about which participants had no previous knowledge or preconceptions - as a proxy for faces. In four experiments (total N = 640) with adult participants we adapt classic AB-A and AB-C renewal paradigms to study appearance-trait learning. Our results indicate that appearance-trait associations are subject to contextual control, and are resistant to counter-stereotypical experience.

Young children are not driven to explore imaginary worlds.

We address Dubourg and Baumard's claim that imaginary worlds are most appealing early in the lifespan when the exploratory drive is highest. Preschool-age children prefer fictions set in the real world, and fantastical information can be difficult for children to represent in real time. We speculate that a drive to explore imaginary worlds may emerge after children acquire substantial real-world skills and knowledge. An account of age effects on fictional preferences should encompass developmental change.

Toward an Account of Intuitive Time.

People hold intuitive theories of the physical world, such as theories of matter, energy, and motion, in the sense that they have a coherent conceptual structure supporting a network of beliefs about the domain. It is not yet clear whether people can also be said to hold a shared intuitive theory of time. Yet, philosophical debates about the metaphysical nature of time often revolve around the idea that people hold one or more "common sense" assumptions about time: that there is an objective "now"; that the past, present, and future are fundamentally different in nature; and that time passes or flows. We empirically explored the question of whether people indeed share some or all of these assumptions by asking adults to what extent they agreed with a set of brief statements about time. Across two analyses, subsets of people's beliefs about time were found consistently to covary in ways that suggested stable underlying conceptual dimensions related to aspects of the "common sense" assumptions described by philosophers. However, distinct subsets of participants showed three mutually incompatible profiles of response, the most frequent of which did not closely match all of philosophers' claims about common sense time. These exploratory studies provide a useful starting point in attempts to characterize intuitive theories of time.

Children's and adults' use of fictional discourse and semantic knowledge for prediction in language processing.

Using real-time eye-movement measures, we asked how a fantastical discourse context competes with stored representations of real-world events to influence the moment-by-moment interpretation of a story by 7-year-old children and adults. Seven-year-olds were less effective at bypassing stored real-world knowledge during real-time interpretation than adults. Our results suggest that children privilege stored semantic knowledge over situation-specific information presented in a fictional story context. We suggest that 7-year-olds' canonical semantic and conceptual relations are sufficiently strongly rooted in statistical patterns in language that have consolidated over time that they overwhelm new and unexpected information even when the latter is fantastical and highly salient.

The seizure-inducing plastic explosive RDX inhibits the α1ß2γ2 GABAA receptor.

OBJECTIVE: Royal demolition explosive (RDX) can induce seizures in wildlife and humans following release into the environment or after voluntary consumption. During the Vietnam War, RDX intoxication was the most common cause of generalized seizures in US service personnel, and in some sections of the armed forces, eating of RDX has continued as "a dare" to this day. After its mechanism of action was long unknown, RDX was recently shown to be a GABAA receptor antagonist. We here determined the GABAA receptor subtype-selectivity of RDX and mapped its functional binding site. METHODS: We used whole-cell patch-clamp to determine the potency of RDX on 10 recombinantly expressed GABAA receptors and mapped the RDX binding site using a combination of Rosetta molecular modeling and site-directed mutagenesis. RESULTS: RDX was found to reversibly inhibit the α1ß2γ2 GABAA receptor with an IC50 of 23 µmol/L (95% CI 15.1-33.3 µmol/L), whereas α4 and α6 containing GABAA receptor combinations were 4-10-fold less sensitive. RDX is binding to the noncompetitive antagonist (NCA) site in the pore. In a molecular model based on the cryo-EM structure of the resting state of the α1ß2γ2 receptor, RDX forms two hydrogen bonds with the threonines at the T6' ring and makes hydrophobic interactions with the valine and alanine in 2' position of the α1 or ß2 subunits. INTERPRETATION: Our findings characterize the mechanism of action of RDX at the atomistic level and suggest that RDX-induced seizures should be susceptible to treatment with GABAA modulating drugs such as benzodiazepines, barbiturates, propofol, or neurosteroids.

Multiorganizational Partnerships: A Mechanism for Increasing the Employment of Internationally Educated Nurses.

BACKGROUND: Internationally educated nurses (IENs) face multiple challenges in entering and integrating into the Canadian workforce. These challenges include getting to know the Canadian culture, nursing accountabilities, professional practice requirements and experience or qualifications deemed not equivalent to the Canadian standard. Hamilton Health Sciences' (HHS') IEN Integration Project has been funded by the Ontario and Canadian governments to support IENs in overcoming these challenges and contribute to the healthcare system. AIM: The aim of this article is to describe a multiorganizational project that prepares IENs for employment in Canadian healthcare. STRATEGY: HHS invited partners in education and immigrant support services to co-design the project. A community collaboration employment model (CCEM) was developed to leverage each partner's strengths in targeted interventions to address the needs of IENs, as identified in focus groups. The interventions pertain to professional practice and accountability in the Canadian healthcare setting, workplace language, communication and selected clinical skills. RESULTS: Between project initiation in 2009 and early 2021, 591 IENs obtained employment. CONCLUSION: Multiorganizational partnerships can help build and sustain a strong nursing workforce, and IENs can fill gaps in care. A needs-based approach and the CCEM increased the likelihood of IEN employment. The ability of the CCEM to engage partners makes it relevant for healthcare organizations.

Spontaneous first impressions emerge from brief training.

People have a strong and reliable tendency to infer the character traits of strangers based solely on facial appearance. In five highly powered and pre-registered experiments, we investigate the relative merits of learning and nativist accounts of the origins of these first impressions. First, we test whether brief periods of training can establish consistent first impressions de novo. Using a novel paradigm with Greebles-a class of synthetic object with inter-exemplar variation that approximates that seen between individual faces-we show that participants quickly learn to associate appearance cues with trustworthiness (Experiments 1 and 2). In a further experiment, we show that participants easily learn a two-dimensional structure in which individuals are presented as simultaneously varying in both trustworthiness and competence (Experiment 3). Crucially, in the final two experiments (Experiments 4 and 5) we show that, once learned, these first impressions occur following very brief exposure (100 ms). These results demonstrate that first impressions can be rapidly learned and, once learned, take on features previously thought to hold only for innate first impressions (rapid availability). Taken together, these results highlight the plausibility of learning accounts of first impressions.

Commentary: Transition of Internationally Educated Nurses into Practice: What We Need to Do to Evolve as an Inclusive Profession over the Next Decade.

Confronted with a pandemic amid a nursing shortage and an aging and culturally diverse patient population, nursing leaders need to explore innovative ways to meet increasing human resource demands to ensure patient safety. Internationally educated nurses (IENs) are an untapped resource to sustain the nursing workforce and provide culturally appropriate care. Based on the authors' lived experience and knowledge in supporting IENs, this commentary highlights the challenges that threaten the integration of IENs into the Canadian nursing workforce. It also recommends practical strategies to address these challenges, guide innovative and sustainable change and unpack systemic barriers to achieve organizational diversity and equity.

An in vivo Cell-Based Delivery Platform for Zinc Finger Artificial Transcription Factors in Pre-clinical Animal Models.

Zinc finger (ZF), transcription activator-like effectors (TALE), and CRISPR/Cas9 therapies to regulate gene expression are becoming viable strategies to treat genetic disorders, although effective in vivo delivery systems for these proteins remain a major translational hurdle. We describe the use of a mesenchymal stem/stromal cell (MSC)-based delivery system for the secretion of a ZF protein (ZF-MSC) in transgenic mouse models and young rhesus monkeys. Secreted ZF protein from mouse ZF-MSC was detectable within the hippocampus 1 week following intracranial or cisterna magna (CM) injection. Secreted ZF activated the imprinted paternal Ube3a in a transgenic reporter mouse and ameliorated motor deficits in a Ube3a deletion Angelman Syndrome (AS) mouse. Intrathecally administered autologous rhesus MSCs were well-tolerated for 3 weeks following administration and secreted ZF protein was detectable within the cerebrospinal fluid (CSF), midbrain, and spinal cord. This approach is less invasive when compared to direct intracranial injection which requires a surgical procedure.

Identification of the Functional Binding Site for the Convulsant Tetramethylenedisulfotetramine in the Pore of the α 2 ß 3 γ 2 GABAA Receptor.

Tetramethylenedisulfotetramine (TETS) is a so-called "caged" convulsant that is responsible for thousands of accidental and malicious poisonings. Similar to the widely used GABA receptor type A (GABAA) antagonist picrotoxinin, TETS has been proposed to bind to the noncompetitive antagonist (NCA) site in the pore of the receptor channel. However, the TETS binding site has never been experimentally mapped, and we here set out to gain atomistic level insights into how TETS inhibits the human α 2 ß 3 γ 2 GABAA receptor. Using the Rosetta molecular modeling suite, we generated three homology models of the α 2 ß 3 γ 2 receptor in the open, desensitized, and closed/resting state. Three different ligand-docking algorithms (RosettaLigand, Glide, and Swissdock) identified two possible TETS binding sites in the channel pore. Using a combination of site-directed mutagenesis, electrophysiology, and modeling to probe both sites, we demonstrate that TETS binds at the T6' ring in the closed/resting-state model, in which it shows perfect space complementarity and forms hydrogen bonds or makes hydrophobic interactions with all five pore-lining threonine residues of the pentameric receptor. Mutating T6' in either the α 2 or ß 3 subunit reduces the IC50 of TETS by ∼700-fold in whole-cell patch-clamp experiments. TETS is thus interacting at the NCA site in the pore of the GABAA receptor at a location that is overlapping but not identical to the picrotoxinin binding site. SIGNIFICANCE STATEMENT: Our study identifies the binding site of the highly toxic convulsant tetramethylenedisulfotetramine (TETS), which is classified as a threat agent by the World Health Organization. Using a combination of homology protein modeling, ligand docking, site-directed mutagenesis, and electrophysiology, we show that TETS is binding in the pore of the α2ß3γ2 GABA receptor type A receptor at the so-called T6' ring, wherein five threonine residues line the permeation pathway of the pentameric receptor channel.

Electrolocation? The evidence for redox-mediated taxis in Shewanella oneidensis.

Shewanella oneidensis is a dissimilatory metal reducing bacterium and model for extracellular electron transfer (EET), a respiratory mechanism in which electrons are transferred out of the cell. In the last 10 years, migration to insoluble electron acceptors for EET has been shown to be nonrandom and tactic, seemingly in the absence of molecular or energy gradients that typically allow for taxis. As the ability to sense, locate, and respire electrodes has applications in bioelectrochemical technology, a better understanding of taxis in S. oneidensis is needed. While the EET conduits of S. oneidensis have been studied extensively, its taxis pathways and their interplay with EET are not yet understood, making investigation into taxis phenomena nontrivial. Since S. oneidensis is a member of an EET-encoding clade, the genetic circuitry of taxis to insoluble acceptors may be conserved. We performed a bioinformatic analysis of Shewanella genomes to identify S. oneidensis chemotaxis orthologs conserved in the genus. In addition to the previously reported core chemotaxis gene cluster, we identify several other conserved proteins in the taxis signaling pathway. We present the current evidence for the two proposed models of EET taxis, "electrokinesis" and flavin-mediated taxis, and highlight key areas in need of further investigation.

Molecular Analysis of the Bloodmeals of Culex spp. Mosquitoes at Natural Habitats in Singapore to Investigate the Potential Risk of Japanese Encephalitis Virus and West Nile Virus Transmission.

Japanese encephalitis virus (JEV) and West Nile virus (WNV) are arboviruses primarily transmitted by Culex spp. mosquitoes. Birds are the primary hosts for JEV and WNV. Recent WNV outbreaks in Europe and United States and their association with migratory birds highlight the importance of understanding the feeding host preference of potential vectors for outbreak preparedness, especially in nonendemic settings. Singapore is nonendemic to JEV and WNV, but is a stopover site for migratory birds of the East Asian-Australasian Flyway. Therefore, we elucidated the feeding host range of Culex spp. mosquitoes captured in four natural (bird) habitats in Singapore from January 2011 to December 2012. We characterized feeding host DNA in field-caught mosquitoes using a PCR sequencing-based assay targeting the mitochondrial gene regions. Of 22,648 mosquitoes captured, 21,287 belonged to the Culex vishnui subgroup. The host DNA analysis showed that mosquitoes from the Cx. vishnui subgroup are opportunistic biters, feeding on a range of birds and mammals. Cx. vishnui subgroup, Culex sitiens and Culex bitaeniorhynchus, was primarily ornithophagic, although they fed opportunistically on mammals, including humans. Culex gelidus and Culex quinquefasciatus, in contrast, fed mainly on mammals. The presence of ornitho- and anthropophilic mosquito vectors and susceptible avian and mammalian hosts poses a risk spill-over transmission of JEV and WNV among humans, should these viruses be introduced through migratory birds and establish persistent transmission in resident birds and animal hosts in Singapore.

The Effect of Tacrolimus Trough Variability on Kidney Transplant Outcomes.

PURPOSE: Variability in tacrolimus levels has been associated with increased rejection, graft loss, and de novo donor-specific antibody (dnDSA) development in kidney transplant recipients (KTRs); however, limited data on alemtuzumab induction or infection exist. We sought to determine the impact of tacrolimus variability in KTRs on dnDSAs, graft outcomes, and infections 3 years posttransplant after alemtuzumab induction. METHODS: Adult KTRs from January 1, 2013, to December 31, 2017, receiving alemtuzumab and tacrolimus-based immunosuppression at a single center were included. Tacrolimus variability was calculated using coefficient of variability (CV), and high CV was defined as ≥30%. Graft and infectious outcomes were assessed between high and low CV groups. RESULTS: Two hundred fourteen KTRs were included. The median tacrolimus CV from 0 to 3 months and from 3 to 12 months was 28.1% and 25.8%, respectively. Recipients with high CV had decreased glomerular filtration rate at 3 and 12 months (67.7 ± 35.48 vs 80.7 ± 29.3, P = .01 and 70.9 ± 35.4 vs 83.3 ± 30.2, P = .015). High CV was also associated with increased cytomegalovirus viremia and disease (19.6% vs 9.3%, P = .046 and 6.4% vs 17.9%, P = .015). No difference in biopsy-proven acute rejection, survival, or dnDSA development at 3 years was observed. CONCLUSIONS: High tacrolimus variability was associated with significantly reduced graft function and increased cytomegalovirus viremia and disease but not biopsy-proven acute rejection, survival, or dnDSA development.

Pain in the Past and Pleasure in the Future: The Development of Past-Future Preferences for Hedonic Goods.

It seems self-evident that people prefer painful experiences to be in the past and pleasurable experiences to lie in the future. Indeed, it has been claimed that, for hedonic goods, this preference is absolute (Sullivan, 2018). Yet very little is known about the extent to which people demonstrate explicit preferences regarding the temporal location of hedonic experiences, about the developmental trajectory of such preferences, and about whether such preferences are impervious to differences in the quantity of envisaged past and future pain or pleasure. We find consistent evidence that, all else being equal, adults and children aged 7 and over prefer pleasure to lie in the future and pain in the past and believe that other people will, too. They also predict that other people will be happier when pleasure is in the future rather than the past but sadder when pain is in the future rather than the past. Younger children have the same temporal preferences as adults for their own painful experiences, but they prefer their pleasure to lie in the past and do not predict that others' levels of happiness or sadness vary dependent on whether experiences lie in the past or the future. However, from the age of 7, temporal preferences were typically abandoned at the earliest opportunity when the quantity of past pain or pleasure was greater than the quantity located in the future. Past-future preferences for hedonic goods emerge early developmentally but are surprisingly flexible.