Unintended Activation Research Articles

DO NOT LOSE SLEEP OVER IT: IMPLANTED BRAIN-COMPUTER INTERFACE FUNCTIONALITY DURING NIGHTTIME IN LATE-STAGE AMYOTROPHIC LATERAL SCLEROSIS.

Brain-computer interfaces (BCIs) hold promise as augmentative and alternative communication technology for people with severe motor and speech impairment (locked-in syndrome) due to neural disease or injury. Although such BCIs should be available 24/7, to enable communication at all times, feasibility of nocturnal BCI use has not been investigated. Here, we addressed this question using data from an individual with amyotrophic lateral sclerosis (ALS) who was implanted with an electrocorticography-based BCI that enabled the generation of click-commands for spelling words and call-caregiver signals. We investigated nocturnal dynamics of neural signal features used for BCI control, namely low (LFB: 10-30Hz) and high frequency band power (HFB: 65-95Hz). Additionally, we assessed the nocturnal performance of a BCI decoder that was trained on daytime data by quantifying the number of unintentional BCI activations at night. Finally, we developed and implemented a nightmode decoder that allowed the participant to call a caregiver at night, and assessed its performance. Power and variance in HFB and LFB were significantly higher at night than during the day in the majority of the nights, with HFB variance being higher in 88% of nights. Daytime decoders caused 245 unintended selection-clicks and 13 unintended caregiver-calls per hour when applied to night data. The developed nightmode decoder functioned error-free in 79% of nights over a period of ±1.5 years, allowing the user to reliably call the caregiver, with unintended activations occurring only once every 12 nights. Reliable nighttime use of a BCI requires decoders that are adjusted to sleep-related signal changes. This demonstration of a reliable BCI nightmode and its long-term use by an individual with advanced ALS underscores the importance of 24/7 BCI reliability. This trial is registered in clinicaltrials.gov under number NCT02224469 (https://clinicaltrials.gov/study/NCT02224469?term=NCT02224469&rank=1). Date of submission to registry: August 21, 2014. Enrollment of first participant: September 7, 2015.

Intelligent Video Analytics for Abnormal Event Detection

Surveillance cameras have become a primary tool for monitoring human movement and preventing unwanted or unintended activities. In today's world, security management professionals increasingly rely on video surveillance to combat crime and mitigate incidents that could adversely affect society. The number of surveillance camera installations has dramatically increased in both the private and public sectors, serving to monitor public activities effectively. Video surveillance is considered one of the most efficient methods for ensuring security. Installing a surveillance camera allows the captured footage to be transmitted to security personnel; however, merely having video footage is insufficient for identifying abnormal activities. To effectively analyze this footage, it is essential to integrate an intelligent system.This paper aims to design and implement an Intelligent Video Analytics Model (IVAM), also known as the Human Object Detection (HOD) method, for analyzing and detecting video-based anomalies and abnormal human activities. IVAM can be deployed alongside surveillance cameras in various public locations such as Institutions, airports, hospitals, shopping malls, and railway stations, allowing for the automatic identification of unusual events. The IVAM was experimented with using MATLAB software, and the results were thoroughly verified. The performance of IVAM was assessed by comparing the obtained results with those from existing approaches, demonstrating that IVAM outperforms contemporary methods concerning accurate anomaly detection, lower error rates, and higher classification accuracy.

Rational Design of High-Efficiency Synthetic Small Regulatory RNAs and Their Application in Robust Genetic Circuit Performance Through Tight Control of Leaky Gene Expression.

Synthetic sRNAs show promise as tools for targeted and programmable gene expression manipulation. However, the design of high-efficiency synthetic sRNAs is a challenging task that necessitates careful consideration of multiple factors. Therefore, this study aims to investigate rational design strategies that significantly and robustly enhance the efficiency of synthetic sRNAs. This is achieved by optimizing the following parameters: the sRNA scaffold, mRNA binding affinity, Hfq protein expression level, and mRNA secondary structure. By utilizing optimized synthetic sRNAs within a positive feedback circuit, we effectively addressed the issue of gene expression leakage─an enduring challenge in synthetic biology that undermines the reliability of genetic circuits in bacteria. Our designed synthetic sRNAs successfully prevented gene expression leakage, thus averting unintended circuit activation caused by initial expression noise, even in the absence of signal molecules. This result shows that high-efficiency synthetic sRNAs not only enable precise gene knockdown for metabolic engineering but also ensure the robust performance of synthetic circuits. The strategies developed here hold significant promise for broad applications across diverse biotechnological fields, establishing synthetic sRNAs as pivotal tools in advancing synthetic biology and gene regulation.

Enhancer-Insulator Pairing Reveals Heterogeneous Dynamics in Long-Distance 3D Gene Regulation

Cells regulate fates and complex body plans using spatiotemporal signaling cascades that alter gene expression. Short DNA sequences, known as enhancers (50–1500 base pairs), help coordinate these cascades by attracting regulatory proteins that enhance the transcription by binding to distal gene promoters. In humans, there are hundreds of thousands of enhancers dispersed across the genome, which poses a challenging coordination task to prevent unintended gene activation. To mitigate this problem, the genome contains insulator elements that block enhancer-promoter interactions. However, there is an open problem with how the insulation works, especially as enhancer-insulator pairs may be separated by millions of base pairs. Based on recent empirical data from Hi-C experiments, this paper proposes a new mechanism that challenges the common paradigm that rests on specific insulator-insulator interactions. Instead, this paper introduces a stochastic looping model where insulators bind weakly to chromatin rather than other insulators. After calibrating the model to experimental data, we use simulations to study the broad distribution of hitting times between an enhancer and a promoter when insulators are present. We find parameter regimes with large differences between average and most probable hitting times. This makes it difficult to assign a typical timescale and hints at highly defocused regulation times. We also map our computational model onto a resetting problem that allows us to derive several analytical results. Besides offering new insights into enhancer-insulator interactions, our paper advances the understanding of gene regulatory networks and causal connections between genome folding and gene activation. Published by the American Physical Society 2024

EEG Analyses of visual cue effects on executed movements

BackgroundIn electroencephalographic (EEG) or electrocorticographic (ECoG) experiments, visual cues are commonly used for timing synchronization but may inadvertently induce neural activity and cognitive processing, posing challenges when decoding self-initiated tasks. New methodTo address this concern, we introduced four new visual cues (Fade, Rotation, Reference, and Star) and investigated their impact on brain signals. Our objective was to identify a cue that minimizes its influence on brain activity, facilitating cue-effect free classifier training for asynchronous applications, particularly aiding individuals with severe paralysis. Results22 able-bodied, right-handed participants aged 18–30 performed hand movements upon presentation of the visual cues. Analysis of time-variability between movement onset and cue-aligned data, grand average MRCP, and classification outcomes revealed significant differences among cues. Rotation and Reference cue exhibited favorable results in minimizing temporal variability, maintaining MRCP patterns, and achieving comparable accuracy to self-paced signals in classification. Comparison with existing methodsOur study contrasts with traditional cue-based paradigms by introducing novel visual cues designed to mitigate unintended neural activity. We demonstrate the effectiveness of Rotation and Reference cue in eliciting consistent and accurate MRCPs during motor tasks, surpassing previous methods in achieving precise timing and high discriminability for classifier training. ConclusionsPrecision in cue timing is crucial for training classifiers, where both Rotation and Reference cue demonstrate minimal variability and high discriminability, highlighting their potential for accurate classifications in online scenarios. These findings offer promising avenues for refining brain-computer interface systems, particularly for individuals with motor impairments, by enabling more reliable and intuitive control mechanisms.

The role of spinal cord neuroanatomy in the variances of epidural spinal recordings

BackgroundSpinal cord stimulation (SCS) has demonstrated multiple benefits in treating chronic pain and other clinical disorders related to sensorimotor dysfunctions. However, the underlying mechanisms are still not fully understood, including how electrode placement in relation to the spinal cord neuroanatomy influences epidural spinal recordings (ESRs). To characterize this relationship, this study utilized stimulation applied at various anatomical sections of the spinal column, including at levels of the intervertebral disc and regions correlating to the dorsal root entry zone.MethodTwo electrode arrays were surgically implanted into the dorsal epidural space of the swine. The stimulation leads were positioned such that the caudal-most electrode contact was at the level of a thoracic intervertebral segment. Intraoperative cone beam computed tomography (CBCT) images were utilized to precisely determine the location of the epidural leads relative to the spinal column. High-resolution microCT imaging and 3D-model reconstructions of the explanted spinal cord illustrated precise positioning and dimensions of the epidural leads in relation to the surrounding neuroanatomy, including the spinal rootlets of the dorsal and ventral columns of the spinal cord. In a separate swine cohort, implanted epidural leads were used for SCS and recording evoked ESRs.ResultsReconstructed 3D-models of the swine spinal cord with epidural lead implants demonstrated considerable distinctions in the dimensions of a single electrode contact on a standard industry epidural stimulation lead compared to dorsal rootlets at the dorsal root entry zone (DREZ). At the intervertebral segment, it was observed that a single electrode contact may cover 20-25% of the DREZ if positioned laterally. Electrode contacts were estimated to be ~0.75 mm from the margins of the DREZ when placed at the midline. Furthermore, ventral rootlets were observed to travel in proximity and parallel to dorsal rootlets at this level prior to separation into their respective sides of the spinal cord. Cathodic stimulation at the level of the intervertebral disc, compared to an ‘off-disc’ stimulation (7 mm rostral), demonstrated considerable variations in the features of recorded ESRs, such as amplitude and shape, and evoked unintended motor activation at lower stimulation thresholds. This substantial change may be due to the influence of nearby ventral roots. To further illustrate the influence of rootlet activation vs. dorsal column activation, the stimulation lead was displaced laterally at ~2.88 mm from the midline, resulting in variances in both evoked compound action potential (ECAP) components and electromyography (EMG) components in ESRs at lower stimulation thresholds.ConclusionThe results of this study suggest that the ECAP and EMG components of recorded ESRs can vary depending on small differences in the location of the stimulating electrodes within the spinal anatomy, such as at the level of the intervertebral segment. Furthermore, the effects of sub-centimeter lateral displacement of the stimulation lead from the midline, leading to significant changes in electrophysiological metrics. The results of this pilot study reveal the importance of the small displacement of the electrodes that can cause significant changes to evoked responses SCS. These results may provide further valuable insights into the underlying mechanisms and assist in optimizing future SCS-related applications.

Emerging techniques for the trace elemental analysis of plants and food-based extracts: A comprehensive review

This comprehensive review investigates the dynamic landscape of trace elemental analysis methodologies applied to plants and food-based extracts. The exploration spans from the inception of techniques to the latest procedures, contributing to heightened precision and sensitivity in elemental detection. According to the WHO, herbal plants and medicine from varied soil compositions serve as crucial therapeutic agents for 70–80 % of the world's population. Yet, their susceptibility to trace element toxicity poses a significant risk to human health. Rising population and increased food demands have led to environmental pollution, contaminating the food chain through unintended activities like industrialization, mining, and pesticide production. The elemental composition of plants and derived extracts is central to comprehending nutritional profiles, evaluating product quality, and ensuring food safety. Methodological advancements, progressing from manual procedures to sophisticated technologies such as X-ray fluorescence (XRF), neutron activation analysis (NAA), inductively coupled plasma optical emission spectroscopy (ICP OES), and inductively coupled plasma mass spectrometry (ICP-MS), are delineated. Recent strides in paper-based electrochemical sensors are highlighted for their distinctive capabilities and elucidation of associated advantages and limitations.Moreover, the review delves into innovative sample preparation methodologies, encompassing microwave-assisted digestion and solid-phase microextraction, to amplify the efficiency of elemental extraction and subsequent analysis. Integrating data analytics and machine learning in elucidating complex elementary datasets is explored, underscoring the potential for heightened accuracy and automation in trace elemental analysis. This review compiles literature data, summarizing sample preparation methods for various herbal parts (roots, soil, stems, bark, fruits, food). Standard protocols from WHO, United States Pharmacopeia-National Formulary (USP-FR), Ayurveda, Yoga and Naturopathy, Unani, Siddha, and Homeopathy (AYUSH) are considered. Lead, Cadmium, Mercury, and Arsenic are the primary toxic elements of concern in herbal medicines. This review furnishes valuable insights tailored for researchers, analysts, and policymakers actively involved in advancing the domain of trace elemental analysis in plants and food-based extract.

Large- and Small-Animal Studies of Safety, Pharmacokinetics, and Biodistribution of Inflammasome-Targeting Nanoligomer in the Brain and Other Target Organs.

Immune malfunction or misrecognition of healthy cells and tissue, termed autoimmune disease, is implicated in more than 80 disease conditions and multiple other secondary pathologies. While pan-immunosuppressive therapies like steroids can offer limited relief for systemic inflammation for some organs, many patients never achieve remission, and such drugs do not cross the blood-brain barrier, making them ineffective for tackling neuroinflammation. Especially in the brain, unintended activation of microglia and astrocytes is hypothesized to be directly or indirectly responsible for multiple sclerosis, amyotrophic lateral sclerosis, Parkinson's disease, and Alzheimer's disease. Recent studies have also shown that targeting inflammasomes and specific immune targets can be beneficial for these diseases. Furthermore, our previous studies have shown targeting NF-κB and NLRP3 through brain penetrant Nanoligomer cocktail SB_NI_112 (abbreviated as NI112) can be therapeutic for several neurodegenerative diseases. Here, we show safety-toxicity studies, followed by pharmacokinetics and biodistribution in small- (mice) and large-animal (dog) studies of this inflammasome-targeting Nanoligomer cocktail NI112. We conducted studies using four different routes of administration: intravenous, subcutaneous, intraperitoneal, and intranasal, and identified the drug concentration over time using inductively coupled plasma mass spectrometry in the blood serum, the brain (including different brain regions), and other target organs such as liver, kidney, and colon. Our results indicate that the Nanoligomer cocktail has a strong safety profile and shows high biodistribution (F ∼ 0.98) and delivery across multiple routes of administration. Further analysis showed high brain bioavailability with a ratio of NI112 in brain tissue to blood serum of ∼30%. Our model accurately shows dose scaling, translation between different routes of administration, and interspecies scaling. These results provide an excellent platform for human clinical translation and prediction of therapeutic dosage between different routes of administration.

Multiplex Prime Editing and PASSIGE TM for Non-Viral Generation of an Allogeneic CAR-T Cell Product

While CAR-T cell therapy represents a major advance in personalized immunotherapy, the autologous nature of commercially available CAR-T products have delayed its broad application beyond a subset of hematological malignancies. In particular, manufacturing autologous CAR-T therapies is costly and time-consuming, and success relies on the fitness of a patient's cells. An allogeneic off-the-shelf CAR-T product could overcome these cell quality and quantity issues and could be accessed on-demand. However, a successful allogeneic CAR-T product will require multiplex gene knockout in addition to stable CAR integration to prevent graft-versus-host disease (GvHD) and graft rejection by the patient's immune system. Strategies for the delivery and expression of CAR transgenes include semi-random integration using lentivirus or transposons, which risk unintended gene disruption or activation. Targeted integration can be achieved using nucleases in combination with a template for homology-directed repair (HDR), but efficiency of this approach is generally low and there are risks associated with the induction of double-strand breaks (DSBs), including p53 activation and chromothripsis. This is further complicated in a situation where multiplex editing is required, as multiple DSBs can lead to gross chromosomal rearrangements. Alternatively, base editors have been used for targeted gene disruption without the induction of DSBs via single nucleotide conversion to disrupt a splice site or introduce a single premature stop (pmSTOP) codon. However, this approach is limited to transition mutations and cannot be used to integrate large genetic cargo, which is required to generate CAR-T cells. Prime Editing (PE) can be used for precise and programmable gene disruption via multiple strategies, including introduction of multiple pmSTOP codons, splice site disruption, frameshift mutations, or deletion of large segments of regulatory or coding sequence. Further, Prime-Assisted Site-Specific Integrase Gene Editing (PASSIGE) can be used to precisely and flexibly integrate large genetic cargo at a specific locus. Together, PE-mediated gene knock out and PASSIGE can be tailored to generate a more broadly applicable, potentially safer, and more effective CAR-T cell product. To evaluate the efficacy of an all-PE non-viral approach to allogeneic CAR-T cell generation, PASSIGE and PE mediated gene knockout were used to generate CD19-CAR-T cells. Multiplex PE precisely disrupted expression of the endogenous T cell receptor alpha constant ( TRAC) and beta-2-microglobulin ( B2M) loci in over 90% of human T cells. Co-delivery of a non-viral DNA donor template with PASSIGE editing components resulted in targeted integration of a 3.5 kb CD19-CAR transgene expression cassette at the TRAC locus in over 60% of the T cells, with no observed impact on T cell viability, phenotype, or functionality. CD19 CAR-T cells generated using PASSIGE show potent anti-tumor activity and cytokine production in response to CD19 + tumor cell lines in vitro and in vivo. The PASSIGE-generated CD19 CAR-T cells reduce tumor burden and prolong survival of mice bearing CD19 + tumors. These results show that a PE platform can be used to generate multiplex edited CAR-T cells without the need for viral vectors and without causing DSBs. This modular, one-step, non-viral delivery Prime Editing platform expands the applicability of T cell therapies for the treatment of tumors and immune diseases.

Task-dependent alteration of beta-band intermuscular coherence is associated with ipsilateral corticospinal tract excitability.

Beta-band (15-30 Hz) synchronization between the EMG signals of active limb muscles can serve as a non-invasive assay of corticospinal tract integrity. Tasks engaging a single limb often primarily utilize one corticospinal pathway, although bilateral neural circuits can participate in goal-directed actions involving multi-muscle coordination and utilization of feedback. Suboptimal utilization of such circuits after CNS injury can result in unintended mirror movements and activation of pathological synergies. Accordingly, it is important to understand how the actions of one limb (e.g., a less-affected limb after strokes) influence the opposite corticospinal pathway for the rehabilitation target. Certain unimanual actions decrease the excitability of the "unengaged" corticospinal tract, presumably to prevent mirror movement, but there is no direct way to predict the extent to which this will occur. In this study, we tested the hypothesis that task-dependent changes in beta-band drives to muscles of one hand will inversely correlate with changes in the opposite corticospinal tract excitability. Ten participants completed spring pinching tasks known to induce differential 15-30 Hz drive to muscles. During compressions, transcranial magnetic stimulation single pulses to the ipsilateral M1 were delivered to generate motor-evoked potentials in the unengaged hand. The task-induced changes in ipsilateral corticospinal excitability were inversely correlated with associated changes in EMG-EMG coherence of the task hand. These results demonstrate a novel connection between intermuscular coherence and the excitability of the "unengaged" corticospinal tract and provide a springboard for further mechanistic studies of unimanual tasks of varying difficulty and their effects on neural pathways relevant to rehabilitation.

Ordeal by innocence in the big‐data era: Intended data breach disclosure, unintended real activities manipulation

AbstractWe demonstrate an unintended consequence of mandatory disclosure of data breaches: the distortion of firms' real business activities. Employing the staggered adoption of data breach disclosure laws across various US states, we show that mandatory disclosure exacerbates CEOs' real earnings manipulation through production and operation management, which is more pronounced for firms of which the outbreak of data breaches is more of a concern and under stronger short‐term market pressure. The law adoption is associated with higher stock price crash risk and fewer patenting activities. Our findings reveal side effects of certain customer‐protection regulations in view of dampened information quality.

Large-scale prediction of adverse drug reactions-related proteins with network embedding.

Adverse drug reactions (ADRs) are a major issue in drug development and clinical pharmacology. As most ADRs are caused by unintended activity at off-targets of drugs, the identification of drug targets responsible for ADRs becomes a key process for resolving ADRs. Recently, with the increase in the number of ADR-related data sources, several computational methodologies have been proposed to analyze ADR-protein relations. However, the identification of ADR-related proteins on a large scale with high reliability remains an important challenge. In this article, we suggest a computational approach, Large-scale ADR-related Proteins Identification with Network Embedding (LAPINE). LAPINE combines a novel concept called single-target compound with a network embedding technique to enable large-scale prediction of ADR-related proteins for any proteins in the protein-protein interaction network. Analysis of benchmark datasets confirms the need to expand the scope of potential ADR-related proteins to be analyzed, as well as LAPINE's capability for high recovery of known ADR-related proteins. Moreover, LAPINE provides more reliable predictions for ADR-related proteins (Value-added positive predictive value = 0.12), compared to a previously proposed method (P < 0.001). Furthermore, two case studies show that most predictive proteins related to ADRs in LAPINE are supported by literature evidence. Overall, LAPINE can provide reliable insights into the relationship between ADRs and proteomes to understand the mechanism of ADRs leading to their prevention. The source code is available at GitHub (https://github.com/rupinas/LAPINE) and Figshare (https://figshare.com/articles/software/LAPINE/21750245) to facilitate its use. Supplementary data are available at Bioinformatics online.

Abstract 2821: Incorporation of intrinsic checkpoint blockade enhances functionality of multigenic autologous UltraCAR-T® cells manufactured using non-viral gene delivery and rapid manufacturing process

Abstract Current chimeric antigen receptor (CAR)-T manufacturing utilizes viral vectors and extensive ex vivo expansion at large central facilities leading to an exhausted CAR-T phenotype, high costs and long vein-to-vein times. While allogeneic CAR-T can reduce delays in patient treatment, they require extensive manipulation of donor cells, severe lymphodepletion and demonstrate short persistence limiting their therapeutic window. The UltraCAR-T® platform is designed to overcome these limitations by utilizing non-viral multigene delivery and a rapid, decentralized manufacturing process without ex vivo activation or expansion of T cells. Patient’s own T cells are collected and manufactured at the medical center and re-infused one day after gene transfer. Here we describe the next generation UltraCAR-T platform that addresses the inhibitory tumor microenvironment by incorporating a novel mechanism for intrinsic downregulation of one or more checkpoint inhibitor (CPI) genes. Our design achieves intrinsic CPI blockade without gene editing and is aimed at avoiding systemic toxicity and the high cost of combining CPI antibodies. Next generation UltraCAR-T cells simultaneously express CAR, membrane bound IL-15 (mbIL15), kill switch, and incorporate intrinsic CPI blockade. To illustrate the ability of this platform, we designed exemplary non-viral transposons to generate UltraCAR-T cells against multiple tumor targets incorporating intrinsic blockade of either one (PD-1) or two (PD-1 and TIGIT) CPI genes. Healthy donor T cells were transfected using the UltraPorator™ electroporation system to manufacture UltraCAR-T cells without ex vivo activation or expansion. The co-expression of CAR, mbIL15 and kill switch was confirmed by flow cytometry and western blot. Activated UltraCAR-T showed significant reduction in CPI gene expression compared to control CAR-T cells lacking the CPI blockade and did not show unintended off-target activity. Downregulation of CPI gene(s) on UltraCAR-T enhanced cytotoxicity and inflammatory cytokine production, especially at low effector to target (E:T) cell ratios, when co-cultured with PD-L1+/CD155+ tumor cells. Single-cell cytokine proteomics showed significant increase in polyfunctionality of UltraCAR-T with intrinsic downregulation of CPI gene(s). In vivo, a single infusion of receptor tyrosine kinase-like orphan receptor 1 (ROR1)-specific UltraCAR-T with intrinsic PD-1 blockade resulted in rapid expansion, an increase in preferred T cell memory (TSCM/TCM) populations, and significantly improved overall survival of ROR1+ PD-L1+ tumor bearing mice. These preclinical data highlight the improved efficacy of incorporating intrinsic CPI blockade in UltraCAR-T cells using non-viral gene delivery and an established rapid, decentralized manufacturing process. Citation Format: Tim Chan, Cheryl Bolinger, Sean Scott, Mengyan Du, Carol Poortman, Byron Koenitzer, Taranjit Athwal, Lindsey Shepard, R. Daniel Slone, Shourik Dutta, Steven Zilko, James M. Dunleavey, Giorgio Zenere, Jacques Plummer, Bernward Klocke, Christian Zinser, Shamim Ahmad, Douglas E. Brough, Rutul R. Shah, Helen Sabzevari. Incorporation of intrinsic checkpoint blockade enhances functionality of multigenic autologous UltraCAR-T® cells manufactured using non-viral gene delivery and rapid manufacturing process [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 2821.

Human-in-the-loop optimization of visual prosthetic stimulation

Objective. Retinal prostheses are a promising strategy to restore sight to patients with retinal degenerative diseases. These devices compensate for the loss of photoreceptors by electrically stimulating neurons in the retina. Currently, the visual function that can be recovered with such devices is very limited. This is due, in part, to current spread, unintended axonal activation, and the limited resolution of existing devices. Here we show, using a recent model of prosthetic vision, that optimizing how visual stimuli are encoded by the device can help overcome some of these limitations, leading to dramatic improvements in visual perception. Approach. We propose a strategy to do this in practice, using patients’ feedback in a visual task. The main challenge of our approach comes from the fact that, typically, one only has access to a limited number of noisy responses from patients. We propose two ways to deal with this: first, we use a model of prosthetic vision to constrain and simplify the optimization. We show that, if one knew the parameters of this model for a given patient, it would be possible to greatly improve their perceptual performance. Second we propose a preferential Bayesian optimization to efficiently learn these model parameters for each patient, using minimal trials. Main results. To test our approach, we presented healthy subjects with visual stimuli generated by a recent model of prosthetic vision, to replicate the perceptual experience of patients fitted with an implant. Our optimization procedure led to significant and robust improvements in perceived image quality, that transferred to increased performance in other tasks. Significance. Importantly, our strategy is agnostic to the type of prosthesis and thus could readily be implemented in existing implants.

Identification of 2-Pyridinylindole-Based Dual Antagonists of Toll-like Receptors 7 and 8 (TLR7/8).

The toll-like receptors (TLRs) play key roles in activation of the innate immune system. Aberrant activation of TLR7 and TLR8 pathways can occur in the context of autoimmune disorders due to the elevated presence and recognition of self-RNA as activating ligands. Control of this unintended activation via inhibition of TLR7/8 signaling holds promise for the treatment of diseases such as psoriasis, arthritis, and lupus. Optimization of a 2-pyridinylindole series of compounds led to the identification of potent dual inhibitors of TLR7 and TLR8, which demonstrated good selectivity against TLR9 and other family members. The in vitro characterization and in vivo evaluation in rodent pharmacokinetic/pharmacodynamic and efficacy studies of BMS-905 is detailed, along with structural information obtained through X-ray cocrystallographic studies.

Spacer2PAM: A computational framework to guide experimental determination of functional CRISPR-Cas system PAM sequences.

RNA-guided nucleases from CRISPR-Cas systems expand opportunities for precise, targeted genome modification. Endogenous CRISPR-Cas systems in many prokaryotes are attractive to circumvent expression, functionality, and unintended activity hurdles posed by heterologous CRISPR-Cas effectors. However, each CRISPR-Cas system recognizes a unique set of protospacer adjacent motifs (PAMs), which requires identification by extensive screening of randomized DNA libraries. This challenge hinders development of endogenous CRISPR-Cas systems, especially those based on multi-protein effectors and in organisms that are slow-growing or have transformation idiosyncrasies. To address this challenge, we present Spacer2PAM, an easy-to-use, easy-to-interpret R package built to predict and guide experimental determination of functional PAM sequences for any CRISPR-Cas system given its corresponding CRISPR array as input. Spacer2PAM can be used in a ‘Quick’ method to generate a single PAM prediction or in a ‘Comprehensive’ method to inform targeted PAM libraries small enough to screen in difficult to transform organisms. We demonstrate Spacer2PAM by predicting PAM sequences for industrially relevant organisms and experimentally identifying seven PAM sequences that mediate interference from the Spacer2PAM-informed PAM library for the type I-B CRISPR-Cas system from Clostridium autoethanogenum. We anticipate that Spacer2PAM will facilitate the use of endogenous CRISPR-Cas systems for industrial biotechnology and synthetic biology.

ERK inactivation enhances stemness of NSCLC cells via promoting Slug-mediated epithelial-to-mesenchymal transition.

Rationale: The mitogen-activated protein kinase pathway (MAPK) is one of the major cancer-driving pathways found in non-small cell lung cancer (NSCLC) patients. ERK inhibitors (ERKi) have been shown to be effective in NSCLC patients with MAPK pathway mutations. However, like other MAPK inhibitors, ERKi rarely confers complete and durable responses. The mechanism of tumor relapse after ERKi treatment is yet defined.Methods: To best study the mechanism of tumor relapse after ERK inhibitor treatment in NSCLC patients, we treated various NSCLC cell lines and patient-derived xenograft (PDX) with ERK inhibitors and evaluated the enrichment of cancer stem cell (CSC) population. We then performed a Next-generation sequencing (NGS) to identify potential pathways that are responsible for the CSC enrichment. Further, the involvement of specific pathways was examined using molecular and cellular methods. Finally, we investigated the therapeutic benefits of ERKi treatment combined with JAK/STAT pathway inhibitor using cellular and xenograft NSCLC models.Results: We found that ERKi treatment expands the CSC population in NSCLC cells through enhanced epithelial-to-mesenchymal transition (EMT)-mediated cancer cell dedifferentiation. Mechanistically, ERK inactivation induces EMT via pSTAT3-mediated upregulation of Slug, in which, upregulation of miR-204 and downregulation of SPDEF, a transcription repressor of Slug, are involved. Finally, the JAK/STAT pathway inhibitor Ruxolitinib blocks the ERK inactivation-induced EMT and CSC expansion, as well as the tumor progression in xenograft models after ERKi treatment.Conclusions: This study revealed a potential tumor relapse mechanism of NSCLC after ERK inhibition through the unintended activation of the EMT program, ascertained the pSTAT-miR-204-SPDEF-Slug axis, and provided a promising combination inhibitor approach to prevent tumor relapse in patients.

The mineralocorticoid receptor in skin disease.

The mineralocorticoid receptor (MR or NR3C2) is expressed in all types of cells from the different skin compartments. The binding and activation by glucocorticoids has a higher affinity than that on the closely related glucocorticoid receptor (GR or NR3C1). As both corticosteroid receptors are co-express in the skin and considering the therapeutic relevance of glucocorticoids to combat skin inflammatory diseases, it was proposed that several of the major side effects of topical glucocorticoids, such as skin atrophy and delayed wound healing, were due to unintended activation of the MR. Indeed, cutaneous MR blockade using genetic and pharmacological approaches in mice and human reduced corticosteroid-associated skin atrophy in conditions of endogenous and pharmacological glucocorticoid excess. Although data support the safety of topical MR antagonists combined with glucocorticoid, it is crucial to address the efficacy of treatment in skin inflammatory conditions and its impact on the overall metabolism. LINKED ARTICLES: This article is part of a themed issue on Emerging Fields for Therapeutic Targeting of the Aldosterone-Mineralocorticoid Receptor Signaling Pathway. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v179.13/issuetoc.

Stress factor human activities

Abstract. Activities of future humans can influence the safety of a closed repository. The German safety requirements for a final repository for high-level radioactive waste (EndlSiAnfV, 2020) therefore stipulate that developments induced by future human activities which may become relevant for the safety of the repository system must be taken into account when optimising the repository system and verifying its robustness. The focus here is on inadvertent human intrusion into the repository. Developments that can be induced by present human activities serve as reference developments. Dealing with future human activities is challenging. The uncertainties about anthropogenic developments taking place in the 1 million-year assessment period are overwhelming and can hardly be reduced. Moreover, knowledge about human activities in the future cannot be empirically verified, so that it becomes difficult to differentiate between knowledge and mere opinion (Grunwald, 2007, p. 57). Developments in future human activities are the subject of futures research. In the interdisciplinary experiment “Stress factor human activities”, it was therefore investigated whether findings can be derived from futurology sources that might be useful for optimising the repository system and verifying its robustness. Based on potential impacts on the sealed repository, drivers and trends, future narratives, findings and ideas from technical literature and science fiction as well as experiences in the field of radioactive waste management, 25 scenarios of future human activities that may influence the safety of a repository were derived. The spectrum of scenarios ranges from “drift into failure” in uses of the geological subsurface to attacks targeting the repository. It includes biological, chemical and physical impacts on the repository. In addition to direct impacts, those that occur slowly and possibly unnoticed are also addressed. From today's point of view, climate change and the endeavour to open up new reserves of raw materials and energy are important drivers. The distinction between intended and unintended activities is often not clear-cut. The experiment confirms that scenario development is ridden with prerequisites and is necessarily interdisciplinary. Different methodological approaches have to be combined, prerequisites and assumptions have to be clearly identified. Due to the limited time horizon of futures research, it is necessary to continually adapt and update scenarios of future human activities in the sense of a “learning process” with new findings and developments. Although many details remain speculative, scenarios provide a differentiated picture of human activities that may influence the safety of the final repository from the current perspective. Patterns are emerging that indicate how human activities could be incorporated into the optimisation of a repository and the verification of its robustness. The results of the interdisciplinary study “Stress factor human activities” therefore lead to the conclusion that futures research can contribute to optimising the long-term safety of a final repository (Eckhardt, 2021).

Ipsilateral and contralateral responses following unimanual fatigue with and without illusionary mirror visual feedback.

Illusionary mirror visual feedback alters interhemispheric communication and influences cross-limb interactions. Combining forceful unimanual contractions with the mirror illusion is a convenient way to provoke robust alterations within ipsilateral motor networks. It is unknown, however, if the mirror illusion affects cross-limb fatigability. We examine this concept by comparing the ipsilateral and contralateral handgrip force and electromyographic (EMG) responses following unimanual fatigue with and without illusionary mirror visual feedback. Participants underwent three experimental sessions (mirror, no-mirror, and control), performing a unimanual fatigue protocol with and without illusionary mirror visual feedback. Maximal handgrip force and EMG activity were measured before and after each session for both hands during maximal unimanual and bimanual contractions. The associated EMG activity from the inactive forearm during unimanual contraction was also examined. The novel findings demonstrate greater relative fatigability during bimanual versus unimanual contraction following unimanual fatigue (-31.8% vs. -23.4%, P < 0.01) and the mirror illusion attenuates this difference (-30.3% vs. -26.3%, P = 0.169). The results show no evidence for a cross-over effect of fatigue with (+0.62%, -2.72%) or without (+0.26%, -2.49%) the mirror illusion during unimanual or bimanual contraction. The mirror illusion resulted in significantly lower levels of associated EMG activity in the contralateral forearm. There were no sex differences for any of the measures of fatigability. These results demonstrate that the mirror illusion influences contraction-dependent fatigue during maximal handgrip contractions. Alterations in facilitatory and inhibitory transcallosal drive likely explain these findings.NEW & NOTEWORTHY Illusionary mirror visual feedback is a promising clinical tool for motor rehabilitation, yet many features of its influence on motor output are unknown. We show that maximal bimanual force output is compromised to a greater extent than unimanual force output following unimanual fatigue, yet illusionary mirror visual feedback attenuates this difference. The mirror illusion also reduces the unintended EMG activity of the inactive, contralateral forearm during unimanual contraction.