Active Recruitment Research Articles

Serine proteases mediate leukocyte recruitment and hepatic microvascular injury in the acute phase following extended hepatectomy.

Post-hepatectomy liver failure (PHLF) is the main limitation of extended liver resection. The molecular mechanism and the role of leukocytes in the development of PHLF remain to be unveiled. We aimed to address the impact of serine proteases (SPs) on the acute phase after liver resection by intravitally analyzing leukocyte recruitment and changes in hemodynamics and microcirculation of the liver. C57BL/6 mice undergoing 60% partial hepatectomy were treated with aprotinin (broad-spectrum SP inhibitor), tranexamic acid (plasmin inhibitor), or vehicle. Sham-operated animals served as controls. In vivo fluorescence microscopy was used to quantify leukocyte-endothelial interactions immediately after, as well as 120 min after partial hepatectomy in postsinusoidal venules, along with measurement of sinusoidal perfusion rate and postsinusoidal shear rate. Recruitment of leukocytes, neutrophils, T cells, and parameters of liver injury were assessed in tissue/blood samples. Leukocyte recruitment, sinusoidal perfusion failure rate, and shear rate were significantly increased in mice after 60% partial hepatectomy compared to sham-operated animals. The inhibition of SPs or plasmin significantly attenuated leukocyte recruitment and improved the perfusion rate in the remnant liver. ICAM-1 expression and neutrophil recruitment significantly increased after 60% partial hepatectomy and were strongly reduced by plasmin inhibition. Endothelial activation and leukocyte recruitment in the liver in response to the increment of sinusoidal shear rate were hallmarks in the acute phase after liver resection. SPs mediated leukocyte recruitment and contributed to the impairment of sinusoidal perfusion in an ICAM-1-dependent manner in the acute phase after liver resection.

Micrococcus luteus-derived extracellular vesicles attenuate neutrophilic asthma by regulating miRNAs in airway epithelial cells

Bacterial extracellular vesicles (EVs) have been shown to regulate various pulmonary diseases, but their functions in asthma remain uncertain. To demonstrate the clinical significance of Micrococcus luteus-derived EVs (MlEVs) in asthma, we enrolled 45 asthmatic patients (20 patients with neutrophilic asthma [NA], 25 patients with eosinophilic asthma [EA]) and 40 healthy controls (HCs). When the prevalence of IgG1 and IgG4 specific to MlEVs was evaluated in serum by ELISA, lower levels of MlEV-specific IgG4 (but not IgG1) were noted in asthmatic patients than in HCs. Among asthmatic patients, significantly lower levels of MIEV-specific IgG4 were noted in patients with NA than in those with EA. Moreover, there was a positive correlation between serum MlEV-specific IgG4 levels and FEV1 (%) values. In asthmatic C57BL/6 mice, MlEVs significantly attenuated neutrophilic airway inflammation by reducing the production of IL-1β and IL-17 in bronchoalveolar lavage fluid as well as the number of group 3 innate lymphoid cells (ILC3s) in lung tissues. To clarify the functional mechanism of MlEVs in NA, the effect of MlEVs on airway epithelial cells (AECs) and immune cells was investigated ex vivo. According to microarray analysis, MlEVs upregulated hsa-miR-4517 expression in AECs. Moreover, this miRNA could suppress IL-1β production by monocytes, resulting in the inhibition of ILC3 activation and neutrophil recruitment. These findings suggest that MlEVs could be a novel therapeutic agent for managing unresolved NA by regulating miRNA expression in AECs.

Abstract C099: Examining the role of a whole food, plant-based diet on the gut microbiome, intramuscular fat, adiposity and risk for breast cancer in obese, post-menopausal African American women

Abstract Background: Obese postmenopausal African American (AA) women are at increased risk of breast cancer. Obesity and aging have been associated with changes in the gut microbiota. Plant-based diets, such as soy isoflavone consumption may protect against breast cancer development. Methods: Participant inclusion and exclusion criteria informed active recruitment and sampling methods for study participants. A mixed-methods, lifestyle study was conducted to evaluate adoption of a culturally tailored, whole food, plant-based diet and changes in risk for breast cancer among obese, post-menopausal African American women. In the qualitative phase, we examined acceptability and feasibility of adopting a whole-food, plant-based diet. In the clinical research, we examined the effect of the whole food, plant-based diet on reductions in weight, adiposity, inflammation, circulating estrogen, and gut microbiome profile. Results: In five focus group discussions, 20 African American women described their knowledge of breast cancer risk factors, attitude toward a plant-based diet, experience with making lifestyle changes, and preferences for plant-based alternatives to traditional meat-based meals. Twelve African American women completed the 4-week whole food, plant-based diet feeding study, regular DEXA body composition scans, and provided blood and stool samples analyses. Women in the qualitative phase, reported being pleased with the whole-food, plant-based selections; document adherence to the meals with food tracking; and experience an average of a five pound weight loss. Biomarker examination is underway. Conclusion: This study addresses knowledge gaps in breast cancer risk and behavioral modifications affecting risk among an aging, obese, post-menopausal African American female population. Discussion will be provided describing how the outcomes of adiposity, inflammation, circulating estrogen, and gut microbiome profile are measured. As well, explanation of the relationship between these mechanistic outcomes will be explored. Lessons learned and insight regarding attrition and the number of women who dropped out or declined to participate will be shared. The long-term goal of this program of research will shed light on the mechanisms involved in the gut microbiome, tissue integrity and function with aging, and the contributions of changes to these systems on declining health and function in obese, postmenopausal African American women at risk for breast cancer. Citation Format: Desiree Rivers, Kay Umeakunne, Muhammad Mubasher, Brian Rivers, Tiffany Carson. Examining the role of a whole food, plant-based diet on the gut microbiome, intramuscular fat, adiposity and risk for breast cancer in obese, post-menopausal African American women [abstract]. In: Proceedings of the 15th AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2022 Sep 16-19; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2022;31(1 Suppl):Abstract nr C099.

Gauging indirect stakeholder sentiment towards orphanage tourism on Twitter

ABSTRACT This study examined 106,979 tweets posted by 34,370 unique users from 2010 to 2019 to understand the sentiment of those indirectly involved in orphanage tourism and its change over time. The result from this study indicated that despite recent critiques of orphanage tourism, the conversations on orphanage volunteering were still largely positive. The persistence of positive sentiment toward orphanage tourism was due to fleeting critiques, the low rate of new critics joining the discourse, and the limited engagement of the existing users. The findings implied that there is a need for further anti-orphanage tourism campaigns and active recruitment of orphanage tourism critiques. The current study contributed to the limited literature on orphanage tourism by utilizing social media to extract valuable information about the perceptions of not only the directly involved stakeholders but also those indirectly involved.

Cancer trials as opportunities to serve and learn from individuals with human immunodeficiency virus.

Despite strong recommendations for their inclusion in cancer clinical trials, people living with human immunodeficiency virus continue to be underrepresented or have inadequate data collected. This commentary highlights the deficit and emphasizes the need for active recruitment and data collection.

Blueberry Consumption and Changes in Obesity and Diabetes Mellitus Outcomes: A Systematic Review.

Low-grade inflammation and oxidative stress are key mechanisms involved in obesity and related disorders. Polyphenols from blueberry (BB) and bilberries (BiB) might protect against oxidative damage and inflammation. To summarize the effects of BiB or BB consumption in parameters related to obesity and its comorbidities, a search of the literature was performed in PubMed, Embase, and Cochrane Library repositories to identify all studies that evaluated associations of whole BB or BiB with obesity and associated disorders. Thirty-one studies were eligible for inclusion in this review: eight clinical trials and 23 animal studies. In humans, BB consumption only consistently decreased oxidative stress and improved endothelial function. In rodents, BB or BiB consumption caused positive effects on glucose tolerance, nuclear factor-kappa B (Nf-κb) activity, oxidative stress, and triglyceride (TG) content in the liver and hepatic steatosis. The high content of anthocyanins present in BB and BiB seems to attenuate oxidative stress. The decrease in oxidative stress may have a positive impact on glucose tolerance and endothelial function. Moreover, in rodents, these berries seem to protect against hepatic steatosis, through the decreased accumulation of hepatic TGs. BB and BiB might also attenuate inflammation by decreasing Nf-κb activity and immune cell recruitment into the adipose tissue.

Gender balance and geographical diversity in editorial boards of Geochimica et Cosmochimica Acta and Chemical Geology

Background: Members of editorial boards of academic journals are often considered gatekeepers of knowledge and role models for the academic community. Editorial boards should be sufficiently diverse in the background of their members to facilitate publishing manuscripts representing a wide range of research paradigms, methods, and cultural perspectives. Objectives: To critically evaluate changes in the representation of binary gender and geographic diversity over time on the editorial boards of Chemical Geology and Geochimica et Cosmochimica Acta, flagship geochemistry journals, respectively, from the European Association of Geochemistry and the Geochemical Society – Meteoritical Society partnership. Methods: The composition of editorial boards was ascertained as given in the first issue of each year, over 1965–2021 for Chemical Geology and 1950–2021 for Geochimica et Cosmochimica Acta, and members of the editorial boards were coded for their country of affiliation (the country of origin may have been different) and for their binary gender. Results: Gender parity, limited to men and women, and the number of countries of affiliation increased steadily between the late 1980s and 2021. However, the geographic distribution remained dominated by affiliations from North America and Western Europe. The editor-in-chief or board of editors had a significant impact on the diversity of the editorial boards, and both geographic and gender diversity may evolve with nearly every newly appointed editor. However, the persistently substantial under-representation on editorial boards of affiliations outside North America and Europe is of concern and needs to be the focus of active recruitment and ongoing monitoring. This approach will ensure that traditionally low geographic diversity is increased and maintained in the future. Conclusion: Improving diversity and inclusion of editorial boards of academic journals and strengthening journal and disciplinary reputations are mutually reinforcing. Instituting a rotating editorship with emphasis on embedding broader geographic networks and more equitable international recruitment could ensure sustained and wider geographic representation and gender balance of editorial boards and promote originality and quality of published research, representing our global communities.

Pharmacological Mechanism of the Non-hallucinogenic 5-HT2A Agonist Ariadne and Analogs.

Ariadne is a non-hallucinogenic analog in the phenylalkylamine chemical class of psychedelics that is closely related to an established synthetic hallucinogen, 2,5-dimethoxy-4-methyl-amphetamine (DOM), differing only by one methylene group in the α-position to the amine. Ariadne has been tested in humans including clinical trials at Bristol-Myers Company that indicate a lack of hallucinogenic effects and remarkable therapeutic effects, such as rapid remission of psychotic symptoms in schizophrenics, relaxation in catatonics, complete remission of symptoms in Parkinson's disease (PD), and improved cognition in geriatric subjects. Despite these provocative clinical results, the compound has been abandoned as a drug candidate and its molecular pharmacology remained unknown. Here, we report a detailed examination of the in vitro and in vivo pharmacology of Ariadne and its analogs, and propose a molecular hypothesis for the lack of hallucinogenic effects and the therapeutic potential of this compound class. We also provide a summary of previous clinical and preclinical results to contextualize the molecular signaling data. Our results show that Ariadne is a serotonin 5-HT2 receptor agonist, exhibits modest selectivity over 5-HT1 receptors, has no relevant activity at 5-HT4,5,7 and other aminergic receptors, and no substantial affinity at plasma membrane monoamine transporters. Compared to DOM, Ariadne shows lower signaling potency and efficacy in multiple signaling pathways examined (Gq, G11, and β-arrestin2) coupled to 5-HT2A receptors. We confirmed the shift in signaling for an α-propyl analog and provide a molecular docking rationale for the progressive decrease in signaling potency with the growing length of the α-substituent. Ariadne versus DOM exhibits no apparent change in the relative preference between Gq/11 activation and β-arrestin2 recruitment; instead, there is a small but consistent drop in efficacy in these signaling channels. Ariadne acts as a 5-HT2A agonist in vivo in mice and shows markedly attenuated head twitch response (HTR) in comparison to its hallucinogenic analogs, consistent with previous studies in rabbits, cats, and dogs. Hence, we propose the lower 5-HT2A receptor signaling efficacy of this compound class as an explanatory model for the lack of hallucinogenic effects of Ariadne in humans and the dramatically attenuated hallucinosis-like effects in animals (5-HT2A signaling efficacy hypothesis). In terms of reverse translation of the noted clinical therapeutic effects, we used an auxilin knockout model of Parkinson's disease where Ariadne rescued severe motor deficits in this mouse line, on par with the effects of l-DOPA, a notable finding considering Ariadne's lack of activity at dopamine receptors and transporters. Ariadne emerges as a prototype of a new drug class, non-hallucinogenic 5-HT2A agonists, with considerable therapeutic potential across psychiatric and neurological indications.

In situ vaccination with cowpea mosaic virus elicits systemic antitumor immunity and potentiates immune checkpoint blockade

BackgroundIn situ vaccination (ISV) is a cancer immunotherapy strategy in which immunostimulatory reagents are introduced directly into a tumor to stimulate antitumor immunity both against the treated tumor and systemically...

Macropinocytosis of APP is controlled by a cascade of regulatory proteins

AbstractBackgroundThe internalization of the Amyloid Precursor Protein (APP) is recognized as an important step in beta‐amyloid (Aβ) production. We have discovered that macropinocytosis of cell surface APP leads to rapid transport directly to lysosomes, bypassing early and late endosomes. This process leads to APP cleavage to Aβ and is regulated in part by Arf6.Here, we hypothesize that the binding/cross‐linking of APP at the cell surface recruits Fe65 and results in the recruitment and activation of Arf6. Arf6 then activates downstream small GTPases that are known to regulate macropinocytosis in non‐neuronal cells (including Rac1, Cdc42 and RhoA) leading to APP internalization by macropinocytosis.MethodNeuro2a neuronal cells are transfected with fluorescent‐tagged proteins including Fe65, Rac1, Cdc42 and RhoA. Membrane PI(4,5)P2, a marker for membrane ruffling and the initiation of macropinocytosis, is visualized using fluorescent‐tagged marker known as PLCdelta‐PH. Cells are cultured in 37°C confocal dishes on the stage of a Leica SP8 confocal microscope. Fluorescent‐labelled anti‐APP antibodies are added to bind/cross‐link APP at the plasma membrane. We then use confocal microscopy/live cell imaging to visualize the recruitment of the regulatory proteins in real time.ResultUpon anti‐APP antibody binding, we can see APP recruited to sites on the plasma membrane enriched with PI(4,5)P2 and the plasma membrane begins to ruffle. We see Fe65 rapidly recruited to the membrane, along with active Arf6 at these sites of membrane ruffling. We also observe Rac1, Cdc42 and RhoA recruited to these regions. APP at these sites is then observed to rapidly internalize to lysosomes, demonstrated by its colocalization with the lysosomal marker LAMP1.ConclusionThese results demonstrate the active recruitment of GTPase proteins known to regulate macropinocytosis to the membrane at sites of APP binding/cross‐linking. These regulators could be targeted to modulate APP trafficking and reduce the production of Aβ.

Insights into distinct signaling profiles of the µOR activated by diverse agonists.

Drugs targeting the μ-opioid receptor (μOR) are the most effective analgesics available but are also associated with fatal respiratory depression through a pathway that remains unclear. Here we investigated the mechanistic basis of action of lofentanil (LFT) and mitragynine pseudoindoxyl (MP), two μOR agonists with different safety profiles. LFT, one of the most lethal opioids, and MP, a kratom plant derivative with reduced respiratory depression in animal studies, exhibited markedly different efficacy profiles for G protein subtype activation and β-arrestin recruitment. Cryo-EM structures of μOR-Gi1 complex with MP (2.5 Å) and LFT (3.2 Å) revealed that the two ligands engage distinct subpockets, and molecular dynamics simulations showed additional differences in the binding site that promote distinct active-state conformations on the intracellular side of the receptor where G proteins and β-arrestins bind. These observations highlight how drugs engaging different parts of the μOR orthosteric pocket can lead to distinct signaling outcomes.

Distinct signatures of loss of consciousness in focal impaired awareness versus tonic-clonic seizures.

Loss of consciousness is a hallmark of many epileptic seizures and carries risks of serious injury and sudden death. While cortical sleep-like activities accompany loss of consciousness during focal impaired awareness seizures, the mechanisms of loss of consciousness during focal to bilateral tonic-clonic seizures remain unclear. Quantifying differences in markers of cortical activation and ictal recruitment between focal impaired awareness and focal to bilateral tonic-clonic seizures may also help us to understand their different consequences for clinical outcomes and to optimize neuromodulation therapies. We quantified clinical signs of loss of consciousness and intracranial EEG activity during 129 focal impaired awareness and 50 focal to bilateral tonic-clonic from 41 patients. We characterized intracranial EEG changes both in the seizure onset zone and in areas remote from the seizure onset zone with a total of 3386 electrodes distributed across brain areas. First, we compared the dynamics of intracranial EEG sleep-like activities: slow-wave activity (1-4 Hz) and beta/delta ratio (a validated marker of cortical activation) during focal impaired awareness versus focal to bilateral tonic-clonic. Second, we quantified differences between focal to bilateral tonic-clonic and focal impaired awareness for a marker validated to detect ictal cross-frequency coupling: phase-locked high gamma (high-gamma phased-locked to low frequencies) and a marker of ictal recruitment: the epileptogenicity index. Third, we assessed changes in intracranial EEG activity preceding and accompanying behavioural generalization onset and their correlation with electromyogram channels. In addition, we analysed human cortical multi-unit activity recorded with Utah arrays during three focal to bilateral tonic-clonic seizures. Compared to focal impaired awareness, focal to bilateral tonic-clonic seizures were characterized by deeper loss of consciousness, even before generalization occurred. Unlike during focal impaired awareness, early loss of consciousness before generalization was accompanied by paradoxical decreases in slow-wave activity and by increases in high-gamma activity in parieto-occipital and temporal cortex. After generalization, when all patients displayed loss of consciousness, stronger increases in slow-wave activity were observed in parieto-occipital cortex, while more widespread increases in cortical activation (beta/delta ratio), ictal cross-frequency coupling (phase-locked high gamma) and ictal recruitment (epileptogenicity index). Behavioural generalization coincided with a whole-brain increase in high-gamma activity, which was especially synchronous in deep sources and could not be explained by EMG. Similarly, multi-unit activity analysis of focal to bilateral tonic-clonic revealed sustained increases in cortical firing rates during and after generalization onset in areas remote from the seizure onset zone. Overall, these results indicate that unlike during focal impaired awareness, the neural signatures of loss of consciousness during focal to bilateral tonic-clonic consist of paradoxical increases in cortical activation and neuronal firing found most consistently in posterior brain regions. These findings suggest differences in the mechanisms of ictal loss of consciousness between focal impaired awareness and focal to bilateral tonic-clonic and may account for the more negative prognostic consequences of focal to bilateral tonic-clonic.

A Phase 3 Double-Blind, Randomized, Placebo Controlled Study to Evaluate the Efficacy and Safety of AB-205 Plus Standard of Care (SOC) Versus Placebo Plus Standard of Care in Adults with Lymphoma Undergoing High-Dose Therapy and Autologous Hematopoietic Cell Transplantation (E-CELERATE) (NCT05181540) - Trials in Progress

Myeloablative conditioning or high-dose therapy followed by autologous hematopoietic cell transplantation (HDT-AHCT) is a standard of care (SOC) and potentially curative consolidation therapy for eligible patients diagnosed with aggressive lymphoma. Although effective in chemosensitive patients, HDT-AHCT is associated with severe regimen-related toxicities (SRRTs) that affect quality of life and may increase mortality risk (Olivieri et al., BBMT 2018). SRRTs occur due to the off-target cytotoxicity of HDT that involves damage to the microvasculature and the vascular stem cell niches of multiple organs. SRRTs predominantly affect organs with high cell turnover causing severe complications such as diffuse gastrointestinal (GI) mucositis (oral/GI SRRT) and delayed hematopoietic recovery. Rates and severity of toxicities increase with age, relevant to the demographics of the aging lymphoma population. Importantly, patients identify oral/GI SRRT as one of the most severe adverse effects experienced with HDT-AHCT (Stiff et al., BMT 2001). Notwithstanding, there are limited treatment strategies aimed at preventing SRRTs. AB-205 is an experimental, engineered cell therapy that consists of allogeneic, human umbilical vein endothelial cells transduced with the pro-survival E4ORF1+ gene (E-CEL® cells). The proposed mechanism of action is paracrine in nature consisting of cell expression of reparative angiocrine factors which are thought to accelerate the repair injuries from off-target cytotoxicity of myeloablative conditioning. In a previously presented Phase 1/2 study [NCT03925935] in HCT-eligible patients with lymphoma, AB-205 was well tolerated and resulted in a 9% incidence of early oral/GI SRRT [defined as Grade ≥3 mucositis (M), nausea (N), vomiting (V) or diarrhea (D) in systemic lymphoma subjects conditioned with BEAM and BEAM-alternates (Phase 1/2 data is locked and analysis is in progress). This oral/GI SRRT incidence compared highly favorably to an incidence rate of 41% resulting from a contemporary, retrospective observational study conducted at two phase 1/2 sites (done concurrently during the conduct of the phase 1/2 study). Accordingly, a phase 3, randomized, double-blind, placebo-controlled multi-center study (E-CELERATE) has been initiated to further investigate the efficacy and safety of AB-205 in reducing (preventing) severe toxicities and complications related to off-target cytotoxicity of myeloablative HDT. Approximately 140 patients will be enrolled across 25 sites in the United States. Key inclusion criteria: Age ≥ 40; diagnosis of Hodgkin or non-Hodgkin lymphoma who are candidates for HDT-AHCT with one of the following myeloablative conditioning regimens: carmustine, etoposide, cytarabine, melphalan (BEAM); bendamustine, etoposide, cytarabine, melphalan (BeEAM); achieved complete response (CR) or partial response (PR) prior to planned HDT; ECOG ≤ 2; serum bilirubin ≤ 2 mg/dL; AST, ALT, and alkaline phosphatase < 3 × ULN. Key exclusion criteria: Prior HCT; primary CNS lymphoma; systemic lymphoma with secondary CNS involvement at time of relapse prior to planned HDT-AHCT; active malignancy other than the one for which the subject is undergoing HDT-AHCT; subjects with a known history of HIV. Endpoints: The primary endpoint is complete response defined as the absence of oral/GI SRRT (NCI CTCAE Grade ≥ 3 M, N, V, or D from the time of HDT through Day +21 post-HCT). Secondary endpoints include the duration of oral/GI SRRT; patient-reported symptom burden per MD Anderson Symptom Inventory; duration of febrile neutropenia; time to neutrophil engraftment; incidence of Grade ≥ 3 non-hematologic treatment emergent AEs; absence of SRRT; and survival endpoints: non-relapse mortality, progression free survival and overall survival. Eligible patients will be randomized 1:1 to receive SOC supportive care prophylaxis + AB-205 or placebo intravenously (IV) given on Day 0 within 4 hours of AHCT. 20×106 cells/kg AB-205 IV or an analogous volume of placebo IV will be administered. Randomization will be stratified by lymphoma type (HL/NHL), hematopoietic cell transplantation-comorbidity index, disease status prior to HDT (CR/PR). Daily assessments of oral/GI SRRT will occur from start of HDT to Day +21 post AHCT (Figure 1). Current status: Active recruitment and enrollment ongoing [NCT05181540]. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Call the SIM team – a tale of building a simulation faculty in a small district general hospital

For many years, simulation-based education (SBE) at Walsall Manor Hospital (WMH) was carried by a one-man simulation technician, with intermittent input from department facilitators. Inadvertently creating SBE dis-equality across departments. Studies have demonstrated that formalized SBE plans improve training [1] and clinical outcomes [2]. We aimed to create standardisation and equity in SBE across departments by formulating a SBE training and delivery plan and governance structure at WMH. In the Autumn 2021, WMH started standardising SBE across the Trust in order to improve both the undergraduate and postgraduate standard of education [3]. The team grew to incorporate five multidisciplinary members; SIM technician, SIM lead (consultant), SIM nurse, SIM project support, and SIM technician support. With further expansion to now include speciality simulation leads in emergency medicine (EM) and paediatrics. Currently there is active recruitment for speciality leads in other departments. The SIM staff were appointed already holding simulation education related qualifications and/or experience. In addition, staff attended the University of Stafford foundations in simulation and debriefing courses. The governance process has been developed and implemented around the appointment of simulation speciality staff, formation of simulation courses, and simulation delivery. Furthermore, collection of attendance, feedback forms, certificates of participation and attendance have been made mandatory element of simulation delivery. In addition, there has been internal and external investment in increasing simulation equipment, including paediatric manikins and immersive technology. In seven months, achievements have included: a range of simulation-based training events, the implementation of in-situ simulation in acute medicine, simulated sessions for final year medical students, the development of a simulation Foundation Year 1 and 2 curriculum, successful construction and running of mock royal college of physicians viva examination, multiple medical procedure courses, the re-introduction of Ill Medical Patients’ Acute Care Treatment (IMPACT) course, and simulation sessions for student nurses. SBE activities were delivered by and to a multidisciplinary team. Putting together a simulation team and formalizing the governance process around SBE delivery has increased the number of educational activities for both undergraduate medical/nursing students and postgraduate doctors and allied health care professionals. Feedback scores have been good to excellent and multidisciplinary work in EM has improved. The SIM team will continue to promote, implement, embed, and sustain SBE within the Trust to bigger and bolder activities. We would like to thank and acknowledge the Dinwoodie charitable company for their support. 1. Weaver SJ, Salas E, Lyons R, Lazzara EH, Rosen MA, DiazGranados D, Grim JG, Augenstein JS, Birnbach DJ, King H. Simulation-based team training at the sharp end: A qualitative study of simulation-based team training design, implementation, and evaluation in healthcare. Journal of Emergencies, Trauma and Shock. 2010;3(4):369–377. 2. Houzé-Cerfon CH, Boet S, Marhar F, Saint-Jean M, Geeraerts T. Simulation-based interprofessional education for critical care teams: Concept, implementation and assessment. Presse Medicale (Paris, France: 1983). 2019;48(7–8 Pt 1):780–787. 3. Irwin PM, Brown RA, Butler S. The undergraduate simulation framework: standardising design and delivery’. Higher Education, Skills and Work-Based Learning. 2020;11(2):576–586.

The basolateral amygdala to lateral septum circuit is critical for regulating social novelty in mice.

The lateral septum (LS) is a basal forebrain GABAergic region that is implicated in social novelty. However, the neural circuits and cell signaling pathways that converge on the LS to mediate social behaviors aren't well understood. Multiple lines of evidence suggest that signaling of brain-derived neurotrophic factor (BDNF) through its receptor TrkB plays important roles in social behavior. BDNF is not locally produced in LS, but we demonstrate that nearly all LS GABAergic neurons express TrkB. Local TrkB knock-down in LS neurons decreased social novelty recognition and reduced recruitment of neural activity in LS neurons in response to social novelty. Since BDNF is not synthesized in LS, we investigated which inputs to LS could serve as potential BDNF sources for controlling social novelty recognition. We demonstrate that selectively ablating inputs to LS from the basolateral amygdala (BLA), but not from ventral CA1 (vCA1), impairs social novelty recognition. Moreover, depleting BDNF selectively in BLA-LS projection neurons phenocopied the decrease in social novelty recognition caused by either local LS TrkB knockdown or ablation of BLA-LS inputs. These data support the hypothesis that BLA-LS projection neurons serve as a critical source of BDNF for activating TrkB signaling in LS neurons to control social novelty recognition.

Abstract 12263: Lack of Diversity in Aortic Stenosis Progression Cohorts: A Systematic Review

Introduction: Aortic stenosis (AS) is characterized by highly variable progression rates. Differential rates of AS progression have been proposed to explain apparent disparities in the treatment of severe AS. However, the evidence supporting this claim is uncertain. Methods: We conducted a systematic review of MEDLINE and EMBASE from 1989 to 2022 to identify prospective cohort studies of AS hemodynamic progression. We evaluated the pooled patient demographics, AS severity, baseline and mean annual changes in echocardiographic markers, and prognostic factors. Results: We identified 32 prospective cohort studies. Overall, 11,432 patients were followed for 27 +/- 15 months on average. The average age was 67 +/- 8 years. The average percent female was 36 +/- 14%. Race was reported in only 4 (13%) cohorts; the average proportion of white patients was 93.9 +/- 6.1%. The average baseline hemodynamic severity was consistent with aortic sclerosis, mild AS, moderate AS, and severe AS in 3 (9.4%), 8 (25.0%), 19 (59.3%), and 2 (6.3%) studies, respectively. Overall, average baseline and annual changes in echocardiographic values were: peak aortic jet velocity (Vmax) 3.21 +/- 0.61 m/s (0.20 +/- 0.09 m/s/yr); aortic valve area (AVA) 1.23 +/- 0.31 cm2 (-0.09 +/- 0.05 cm2/yr); and mean aortic valve gradient (MG) 26.2 +/- 7.6 mmHg (3.9 +/- 1.6 mmHg/yr). Age, sex, coronary artery disease, hypertension, diabetes, hyperlipidemia, and smoking were not associated with AS progression in &gt;75% cohorts. Baseline Vmax and MG were associated with greater AS progression in 6 cohorts (75%). Baseline aortic valve calcification (AVC) and AVA were consistently associated with greater AS progression in 9 cohorts (100%). One cohort found that women have a greater ΔMG for a given level of AVC. A univariate analysis of race in one cohort showed that African American ethnicity was associated with lower risk of progression from normal aortic valves to incident AS. Conclusion: There is limited representation of women and ethnic minorities in prospective AS progression cohorts. Active recruitment of diverse populations in these cohorts is needed. The available clinical evidence provides limited support for claims of differential progression as the cause of racial disparities in AS treatment.

Strategies for effective implementation and scale-up of a multi-level co-designed men's health initiative "Sheds for Life" in Irish Men's Sheds.

Sheds for Life is a gender-specific tailored men's health initiative engaging "hard-to-reach" men in the Men's Shed setting in Ireland. It is implemented by multiple stakeholders at individual, provider, organization and systems level and thus multiple contextual factors influence its scalability. This research used established implementation science frameworks to guide participatory research approaches that captured the process and identified facilitators of and barriers to implementation and scale-up. Active recruitment, co-design processes, leadership and stakeholder engagement emerged as key facilitators of implementation. Prominent barriers were institutional capacity and funding. Acceptability, adoption and appropriateness of the initiative were high among stakeholders with sustainability largely contingent on funding and staff resources. Findings make a valuable contribution to knowledge by capturing the process involved in the implementation of a complex multi-level men's health intervention. It provides a "how to" guide of strategies to engage hard-to-reach men with health promotion, the operationalization and application of implementation frameworks in community-based health promotion, and the implementation of health promotion in Men's Sheds. Documented barriers and facilitators that impact implementation of a community-based men's health program are rare and provide a valuable blueprint for practitioners, researchers and policy makers in the field.

A cryptic transactivation domain of EZH2 binds AR and AR's splice variant, promoting oncogene activation and tumorous transformation.

Enhancer of Zeste Homolog 2 (EZH2) and androgen receptor (AR) are crucial chromatin/gene regulators involved in the development and/or progression of prostate cancer, including advanced castration-resistant prostate cancer (CRPC). To sustain prostate tumorigenicity, EZH2 establishes non-canonical biochemical interaction with AR for mediating oncogene activation, in addition to its canonical role as a transcriptional repressor and enzymatic subunit of Polycomb Repressive Complex 2 (PRC2). However, the molecular basis underlying non-canonical activities of EZH2 in prostate cancer remains elusive, and a therapeutic strategy for targeting EZH2:AR-mediated oncogene activation is also lacking. Here, we report that a cryptic transactivation domain of EZH2 (EZH2TAD) binds both AR and AR spliced variant 7 (AR-V7), a constitutively active AR variant enriched in CRPC, mediating assembly and/or recruitment of transactivation-related machineries at genomic sites that lack PRC2 binding. Such non-canonical targets of EZH2:AR/AR-V7:(co-)activators are enriched for the clinically relevant oncogenes. We also show that EZH2TAD is required for the chromatin recruitment of EZH2 to oncogenes, for EZH2-mediated oncogene activation and for CRPC growth in vitro and in vivo. To completely block EZH2's multifaceted oncogenic activities in prostate cancer, we employed MS177, a recently developed proteolysis-targeting chimera (PROTAC) of EZH2. Strikingly, MS177 achieved on-target depletion of bothEZH2's canonical (EZH2:PRC2) and non-canonical (EZH2TAD:AR/AR-V7:co-activators) complexes in prostate cancer cells, eliciting far more potent antitumor effects than the catalytic inhibitors of EZH2. Overall, this study reports a previously unappreciated requirement for EZH2TAD for mediating EZH2's non-canonical (co-)activator recruitment and gene activation functions in prostate cancer and suggests EZH2-targeting PROTACs as a potentially attractive therapeutic for the treatment of aggressive prostate cancerthat rely on the circuits wired by EZH2 and AR.

Activation of γ-globin expression by hypoxia-inducible factor 1α.

Around birth, globin expression in human red blood cells (RBCs) shifts from γ-globin to β-globin, which results in fetal haemoglobin (HbF, α2γ2) being gradually replaced by adult haemoglobin (HbA, α2β2)1. This process has motivated the development of innovative approaches to treat sickle cell disease and β-thalassaemia by increasing HbF levels in postnatal RBCs2. Here we provide therapeutically relevant insights into globin gene switching obtained through a CRISPR-Cas9 screen for ubiquitin-proteasome components that regulate HbF expression. In RBC precursors, depletion of the von Hippel-Lindau (VHL) E3 ubiquitin ligase stabilized its ubiquitination target, hypoxia-inducible factor 1α (HIF1α)3,4, to induce γ-globin gene transcription. Mechanistically, HIF1α-HIF1β heterodimers bound cognate DNA elements in BGLT3, a long noncoding RNA gene located 2.7 kb downstream of the tandem γ-globin genes HBG1 and HBG2. This was followed by the recruitment of transcriptional activators, chromatin opening and increased long-range interactions between the γ-globin genes and their upstream enhancer. Similar induction of HbF occurred with hypoxia or with inhibition of prolyl hydroxylase domain enzymes that target HIF1α for ubiquitination by the VHL E3 ubiquitin ligase. Our findings link globin gene regulation with canonical hypoxia adaptation, provide a mechanism for HbF induction during stress erythropoiesis and suggest a new therapeutic approach for β-haemoglobinopathies.

Host generalists dominate fungal communities associated with alpine knotweed roots: a study of Sebacinales.

Bistorta vivipara is a widespread herbaceous perennial plant with a discontinuous pattern of distribution in arctic, alpine, subalpine and boreal habitats across the northern Hemisphere. Studies of the fungi associated with the roots of B. vivipara have mainly been conducted in arctic and alpine ecosystems. This study examined the fungal diversity and specificity from root tips of B. vivipara in two local mountain ecosystems as well as on a global scale. Sequences were generated by Sanger sequencing of the internal transcribed spacer (ITS) region followed by an analysis of accurately annotated nuclear segments including ITS1-5.8S-ITS2 sequences available from public databases. In total, 181 different UNITE species hypotheses (SHs) were detected to be fungi associated with B. vivipara, 73 of which occurred in the Bavarian Alps and nine in the Swabian Alps–with one SH shared among both mountains. In both sites as well as in additional public data, individuals of B. vivipara were found to contain phylogenetically diverse fungi, with the Basidiomycota, represented by the Thelephorales and Sebacinales, being the most dominant. A comparative analysis of the diversity of the Sebacinales associated with B. vivipara and other co-occurring plant genera showed that the highest number of sebacinoid SHs were associated with Quercus and Pinus, followed by Bistorta. A comparison of B. vivipara with plant families such as Ericaceae, Fagaceae, Orchidaceae, and Pinaceae showed a clear trend: Only a few species were specific to B. vivipara and a large number of SHs were shared with other co-occurring non-B. vivipara plant species. In Sebacinales, the majority of SHs associated with B. vivipara belonged to the ectomycorrhiza (ECM)-forming Sebacinaceae, with fewer SHs belonging to the Serendipitaceae encompassing diverse ericoid–orchid–ECM–endophytic associations. The large proportion of non-host-specific fungi able to form a symbiosis with other non-B. vivipara plants could suggest that the high fungal diversity in B. vivipara comes from an active recruitment of their associates from the co-occurring vegetation. The non-host-specificity suggests that this strategy may offer ecological advantages; specifically, linkages with generalist rather than specialist fungi. Proximity to co-occurring non-B. vivipara plants can maximise the fitness of B. vivipara, allowing more rapid and easy colonisation of the available habitats.