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To predict suicidal ideation one year after moderate to severe traumatic brain injury (TBI).Cross-sectional design with data collected through the prospective, longitudinal TBI Model Systems (TBIMS) network at hospitalization and one year after injury. Participants who completed the Patient Health Questionnaire-9 (PHQ-9) suicide item at year one follow-up (N = 4,328) were included.A gradient boosting machine (GBM) algorithm demonstrated the best performance in predicting suicidal ideation one year after TBI. Predictors were PHQ-9 items (except suicidality), Generalized Anxiety Disorder-7 (GAD-7) items, and a measure of heavy drinking. Results of the 10-fold cross-validation GBM analysis indicated excellent classification performance with an AUC of 0.882. Sensitivity was 0.85, and specificity was 0.77. Accuracy was 0.78 (95% CI: 0.77 - 0.79). Feature importance analyses revealed that depressed mood and guilt were the most important predictors of suicidal ideation, followed by anhedonia, concentration difficulties, and psychomotor disturbance.Overall, depression symptoms were most predictive of suicidal ideation. Despite the limited clinical impact of the present findings, machine learning has potential to improve prediction of suicidal behavior, leveraging electronic health record data, to identify individuals at greatest risk, thereby facilitating intervention and optimization of long-term outcomes following TBI.

Understanding species’ habitat and resource requirements is critical for effective conservation. To assess whether a species is imperiled, data are needed on its range and natural history. In this study, we examined the resource use, including diet and habitat associations, of the Slenderwrist Burrowing Crayfish, Fallicambarus petilicarpus, a narrowly endemic primary burrowing crayfish in southcentral Arkansas. We found that F. petilicarpus has an omnivorous diet, feeding on aquatic invertebrates and detritus. We compared habitat characteristics of sites in the Ouachita drainage that contained F. petilicarpus and sites with other primary burrowing crayfish. Fallicambarus petilicarpus inhabits roadside ditches with low elevation and reduced canopy cover, like other primary burrowing species in the region. Comparisons among burrowing crayfish suggest there may be overlap in environmental niches of species in this region and subsequent competition. Field sampling expanded the known range of F. petilicarpus from ~174 km2 to ~1150 km2. Fallicambarus petilicarpus remains a narrowly endemic species with potential to be threatened by habitat loss and food scarcity caused by urban expansion and climate change.

This study summarises information on the occurrence of Myrmecophilus acervorum in Poland in the years 1851–2022. We collected information about 143 localities: 40 from literature and 103 unpublished. Myrmecophilus acervorum is reported from seven new regions: the Baltic Coast, Białowieża Primeval Forest, Eastern Sudety Mts, Kraków-Wieluń Upland, Podlasie Lowland, Sandomierska Lowland and Trzebnickie Hills. While until 2010 observations were rare (35 localities), in the years 2011–2022 the species was found in 108 new localities. A total of 443 individuals of M. acervorum were observed in Poland, but the sex was identified only for 138, of which 135 were mature females or female nymphs. Males were recorded only historically in the Pomeranian Lakeland. At present, these records should be treated with caution, as they are uncertain.

The need to control the activity and fidelity of CRISPR-associated nucleases has resulted in a demand for inhibitory anti-CRISPR molecules. The small-molecule inhibitor discovery platforms available at present are not generalizable to multiple nuclease classes, only target the initial step in the catalytic activity and require high concentrations of nuclease, resulting in inhibitors with suboptimal attributes, including poor potency. Here we report a high-throughput discovery pipeline consisting of a fluorescence resonance energy transfer-based assay that is generalizable to contemporary and emerging nucleases, operates at low nuclease concentrations and targets all catalytic steps. We applied this pipeline to identify BRD7586, a cell-permeable small-molecule inhibitor of SpCas9 that is twofold more potent than other inhibitors identified to date. Furthermore, unlike the reported inhibitors, BRD7586 enhanced SpCas9 specificity and its activity was independent of the genomic loci, DNA-repair pathway or mode of nuclease delivery. Overall, these studies describe a general pipeline to identify inhibitors of contemporary and emerging CRISPR-associated nucleases.

CRISPR/Cas9-based genome-editing therapies are poised to change the clinical outcome for many diseases with validated therapeutic targets awaiting an appropriate delivery system. Recent advances in lipid nanoparticle (LNP) technology make them an attractive platform for the delivery of various forms of CRISPR/Cas9, including the efficient and transient Cas9/gRNA ribonucleoprotein (RNP) complexes. In this study, we initially tested our novel LNP platform by delivering pre-complexed RNPs and template DNA to cultured mouse cortical neurons, and obtained successful ex vivo genome editing. We then directly injected LNP-packaged RNPs and DNA template into the mouse cornea to evaluate in vivo delivery. For the first time, we demonstrated wide-spread genome editing in the cornea using our LNP-RNPs. The ability of our LNPs to transfect the cornea highlights the potential of our novel delivery platform to be used in CRISPR/Cas9-based genome editing therapies of corneal diseases.

Background: Most cancer drugs enter the US market first. US Food and Drug Administration (FDA) approvals of new cancer drugs may influence regulatory decisions in other settings. The study examined whether characteristics of available evidence at FDA approval influenced time-to-marketing authorisation (MA) in Brazil, and price differences between the two countries. Methods: All new FDA-approved cancer drugs from 2010 to 2019 were matched to drugs with MA and prices approved in Brazil by December 2020. Characteristics of main studies, availability of randomised controlled trials (RCTs), overall survival (OS) benefit, added therapeutic benefit, and prices were compared. Findings: Fifty-six FDA-approved cancer drugs with matching indications received a MA at the Brazilian Health Regulatory Agency (Anvisa) after a median of 522 days following US approval (IQR: 351–932). Earlier authorisation in Brazil was associated with availability of RCT (median: 506 vs 760 days, p = 0.031) and evidence of OS benefit (390 vs 543 days, p = 0.019) at FDA approval. At Brazilian marketing authorisation, a greater proportion of cancer drugs had main RCTs (75% vs 60.7%) and OS benefit (42.9% vs 21.4%) than that in the US. Twenty-eight (50%) drugs did not demonstrate added therapeutic benefit over drugs for the same indication in Brazil. Median approved prices of new cancer drugs were 12.9% lower in Brazil compared to the US (adjusted by Purchasing Power Parity). However, for drugs with added therapeutic benefit median prices were 5.9% higher in Brazil compared to the US, while 17.9% lower for those without added benefit. Interpretation: High-quality clinical evidence accelerated the availability of cancer medicines in Brazil. The combination of marketing and pricing authorisation in Brazil may favour the approval of cancer drugs with better supporting evidence, and more meaningful clinical benefit albeit with variable degree of success in achieving lower prices compared to the US. Funding: None.

Copyleft AI with Trusted Enforcement (CAITE) can support an adaptable soft law approach for ethics in AI

Among the polypeptides that comprise the T cell receptor (TCR), only CD3ζ is found in Natural Killer (NK) cells, where it transmits signals from activating receptors such as CD16 and NKp46. NK cells are potent immune cells that recognize target cells through germline-encoded activating and inhibitory receptors. Genetic engineering of NK cells enables tumor-specific antigen recognition and, thus, has a significant promise in adoptive cell therapy. Ectopic expression of engineered TCR components in T cells leads to mispairing with the endogenous components, making a knockout of the endogenous TCR necessary. To circumvent the mispairing of TCRs or the need for knockout technologies, TCR complex expression has been studied in NK cells. In the current study, we explored the cellular processing of the TCR complex in NK cells. We observed that in the absence of CD3 subunits, the TCR was not expressed on the surface of NK cells and vice versa. Moreover, a progressive increase in surface expression of TCR between day three and day seven was observed after transduction. Interestingly, the TCR complex expression in NK92 cells was enhanced with a proteasome inhibitor (bortezomib) but not a lysosomal inhibitor (chloroquine). Additionally, we observed that the TCR complex was functional in NK92 cells as measured by estimating CD107a as a degranulation marker, IFNγ cytokine production, and killing assays. NK92 cells strongly degranulated when CD3ε was engaged in the presence of TCR, but not when only CD3 was overexpressed. Therefore, our findings encourage further investigation to unravel the mechanisms that prevent the surface expression of the TCR complex.

Funding Acknowledgements Type of funding sources: None. Introduction Anxiety, depression, and post-traumatic stress disorder (PTSD) among out-of-hospital cardiac arrest (OHCA) survivors may impact their long-term recovery and quality of life. Gaining knowledge on sex differences in psychological consequences following cardiac arrest is important to explore strategies aiming to reduce these symptoms. Purpose To explore sex differences in anxiety, depression and PTSD levels after discharge among OHCA survivors hospitalized at a cardiovascular intensive care unit. Methods This was a prospective observational study in OHCA survivors that attended a follow-up after 4 months, (median, IQR 3-8). Presence of symptoms of anxiety and depression was measured using Hospital Anxiety and Depression Scale (HADS), range 0-21. Scores between 8 to 10 suggest the possible presence of a mood disorder whereas scores of 11 and above indicate the probable presence of a mood disorder. Symptoms of PTSD were measured with PTSD Checklist for DSM-5 (PCL-5), a 20-item self-report measure that assesses symptoms of PTSD, range 0-80. Results Between 2016 and 2021, 245 consecutive OHCA survivors admitted in coma (44 women, 18% and 201 men, 82%) completed the survey. Mean values measured on HADS were 2.7 ±3.2 for depression and 4.8±3.9 for anxiety. Depression and anxiety levels were significantly higher in women (3.3±3.4 and 6.1±3.8), respectively, compared to men (2.6±3.2 and 4.5± 3.9), respectively (p Conclusion Symptoms of anxiety and PTSD are frequent in OHCA survivors and female cardiac arrest survivors report significantly higher symptom levels of anxiety, depression, and PTSD compared to men. These results underline the presence of long-term symptoms in these patients and support further research in sex-specific approaches to alleviating these symptoms in patients surviving OHCA.

Natural killer (NK) cells recognize virally infected cells and tumors. NK cell function depends on balanced signaling from activating receptors, recognizing products from tumors or viruses, and inhibitory receptors (such as KIR/Ly49), which recognize major histocompatibility complex class I (MHC-I) molecules. KIR/Ly49 signaling preserves tolerance to self but also conveys reactivity toward MHC-I–low target cells in a process known as NK cell education. Here, we found that NK cell tolerance and education were determined by the subcellular localization of the tyrosine phosphatase SHP-1. In mice lacking MHC-I molecules, uneducated, self-tolerant Ly49A + NK cells showed accumulation of SHP-1 in the activating immune synapse, where it colocalized with F-actin and the signaling adaptor protein SLP-76. Education of Ly49A + NK cells by the MHC-I molecule H2D d led to reduced synaptic accumulation of SHP-1, accompanied by augmented signaling from activating receptors. Education was also linked to reduced transcription of Ptpn6 , which encodes SHP-1. Moreover, synaptic SHP-1 accumulation was reduced in NK cells carrying the H2D d -educated receptor Ly49G2 but not in those carrying the noneducating receptor Ly49I. Colocalization of Ly49A and SHP-1 outside of the synapse was more frequent in educated compared with uneducated NK cells, suggesting a role for Ly49A in preventing synaptic SHP-1 accumulation in NK cell education. Thus, distinct patterning of SHP-1 in the activating NK cell synapse may determine NK cell tolerance.

PDF file 601K, Supplementary Methods. Figure S1. The NUP98-PHF23 (NP23) transgene and peripheral blood (PB) characteristics of NP23 mice. Figure S2. NP23 AML and pre-T LBL. Figure S3. NP23 bi-clonal AML/pre-T LBL, B-ALL and erythroid leukemias. Figure S4.Impaired hematopoietic development in NP23 progenitor cell populations. Figure S5. Validation of differential gene expression in NP23 tissues. Figure S6. NP23 cells are sensitive to disulfiram, an inhibitor of PHD domain H3K4me3 binding

XLSX file 368K, Table S1. NP23 Diagnoses. Table S2. Notch1 mutations. Table S3. Leukemic fold change (gene expression). Table S4. Leukemic unions, upregulated genes. Table S5. Leukemic unions, downregulated genes. Table S6. NP23 fold change, gene expression. Table S7. T-cell line ChIP-seq. Table S8. 961C cell line ChIP-seq. Table S9. NP23+H3K4me3 loci versus upregulated genes. Table S10. PCR primers

Robust, generalizable approaches to identify compounds efficiently with undesirable mechanisms of action in complex cellular assays remain elusive. Such a process would be useful for hit triage during high-throughput screening and, ultimately, predictive toxicology during drug development. We generated cell painting and cellular health profiles for 218 prototypical cytotoxic and nuisance compounds in U-2 OS cells in a concentration-response format. A diversity of compounds causing cellular damage produced bioactive cell painting morphologies, including cytoskeletal poisons, genotoxins, nonspecific electrophiles, and redox-active compounds. Further, we show that lower quality lysine acetyltransferase inhibitors and nonspecific electrophiles can be distinguished from more selective counterparts. We propose that the purposeful inclusion of cytotoxic and nuisance reference compounds such as those profiled in this Resource will help with assay optimization and compound prioritization in complex cellular assays like cell painting.

This is the first record of the European yellow sac spider Cheiracanthium punctorium (Villers, 1789) from the Lublin Upland (SE Poland). A dozen or so silk nests containing these spiders were found among goldenrod stems (Solidago sp.) on a dry meadow near Chodel, a village in the valley of the River Chodelka on 8 October 2021.

This proceedings article presents actionable research targets on the basis of the presentations and discussions at the 2nd Curing Coma National Institutes of Health (NIH) symposium held from May 3 to May 5, 2021. Here, we summarize the background, research priorities, panel discussions, and deliverables discussed during the symposium across six major domains related to disorders of consciousness. The six domains include (1) Biology of Coma, (2) Coma Database, (3) Neuroprognostication, (4) Care of Comatose Patients, (5) Early Clinical Trials, and (6) Long-term Recovery. Following the 1st Curing Coma NIH virtual symposium held on September 9 to September 10, 2020, six workgroups, each consisting of field experts in respective domains, were formed and tasked with identifying gaps and developing key priorities and deliverables to advance the mission of the Curing Coma Campaign. The highly interactive and inspiring presentations and panel discussions during the 3-day virtual NIH symposium identified several action items for the Curing Coma Campaign mission, which we summarize in this article.

Type 2 diabetes is marked by progressive β-cell failure, leading to loss of β-cell mass. Increased levels of circulating glucose and free fatty acids associated with obesity lead to β-cell glucolipotoxicity. There are currently no therapeutic options to address this facet of β-cell loss in obese type 2 diabetes patients. To identify small molecules capable of protecting β-cells, we performed a high-throughput screen of 20,876 compounds in the rat insulinoma cell line INS-1E in the presence of elevated glucose and palmitate. We found 312 glucolipotoxicity-protective small molecules (1.49% hit rate) capable of restoring INS-1E viability, and we focused on 17 with known biological targets. 16 of the 17 compounds were kinase inhibitors with activity against specific families including but not limited to cyclin-dependent kinases (CDK), PI-3 kinase (PI3K), Janus kinase (JAK), and Rho-associated kinase 2 (ROCK2). 7 of the 16 kinase inhibitors were PI3K inhibitors. Validation studies in dissociated human islets identified 10 of the 17 compounds, namely, KD025, ETP-45658, BMS-536924, AT-9283, PF-03814735, torin-2, AZD5438, CP-640186, ETP-46464, and GSK2126458 that reduced glucolipotoxicity-induced β-cell death. These 10 compounds decreased markers of glucolipotoxicity including caspase activation, mitochondrial depolarization, and increased calcium flux. Together, these results provide a path forward toward identifying novel treatments to preserve β-cell viability in the face of glucolipotoxicity.