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Background: Leadless cardiac resynchronization therapy (CRT) is an emerging heart failure treatment. An implanted electrode delivers lateral or septal endocardial left ventricular (LV) pacing (LVP) upon detection of a right ventricular (RV) pacing stimulus from a coimplanted device, thus generating biventricular pacing (BiVP). Electrical efficacy data regarding this therapy, particularly leadless LV septal pacing (LVSP) for potential conduction system capture, are limited. Objectives: The purpose of this study was to evaluate the acute performance of leadless CRT using electrocardiographic imaging (ECGi) and assess the optimal pacing modality (OPM) of LVSP on the basis of RV and LV activation. Methods: Ten WiSE-CRT recipients underwent an ECGi study testing: RV pacing, BiVP, LVP only, and LVP with an optimized atrioventricular delay (LV-OPT). BiV, LV, and RV activation times (shortest time taken to activate 90% of the ventricles [BIVAT-90], shortest time taken to activate 95% of the LV, and shortest time taken to activate 90% of the RV) plus LV and BiV dyssynchrony index (standard deviation of LV activation times and standard deviation of all activation times) were calculated from reconstructed epicardial electrograms. The individual OPM yielding the greatest improvement from baseline was determined. Results: BiVP generated a 23.7% improvement in BiVAT-90 (P = .002). An improvement of 43.3% was observed at the OPM (P = .0001), primarily through reductions in shortest time taken to activate 90% of the RV. At the OPM, BiVAT-90 improved in patients with lateral (43.3%; P = .0001; n = 5) and septal (42.4%; P = .009; n = 5) LV implants. The OPM varied by individual. LVP and LV-OPT were mostly superior in patients with LVSP, and in those with sinus rhythm and left bundle branch block (n = 4). Conclusion: Leadless CRT significantly improves acute ECGi-derived activation and dyssynchrony metrics. Using an individualized OPM improves efficacy in selected patients. Effective LVSP is feasible, with fusion pacing at LV-OPT mitigating the potential deleterious effects on RV activation.

Abstract Botulinum neurotoxins (BoNTs) are the most potent toxins known and are used to treat an increasing number of medical disorders. All BoNTs are naturally co-expressed with a protective partner protein (NTNH) with which they form a 300 kDa complex, to resist acidic and proteolytic attack from the digestive tract. We have previously identified a new botulinum neurotoxin serotype, BoNT/X, that has unique and therapeutically attractive properties. We present the cryo-EM structure of the BoNT/X-NTNH/X complex and the crystal structure of the isolated NTNH protein. Unexpectedly, the BoNT/X complex is stable and protease-resistant at both neutral and acidic pH and disassembles only in alkaline conditions. Using the stabilizing effect of NTNH, we isolated BoNT/X and showed that it has very low potency both in vitro and in vivo. Given the high catalytic activity and translocation efficacy of BoNT/X, low activity of the full toxin is likely due to the receptor-binding domain, which presents very weak ganglioside binding and exposed hydrophobic surfaces.

The thermo-osmotic energy conversion (TOEC) process harnesses low-grade waste heat for electricity generation. Key to TOEC is selecting membrane materials, with polyvinylidene fluoride (PVDF) and polytetrafluoroethylene (PTFE) being common choices. This study provides the first life cycle assessment (LCA) of PTFE and PVDF membranes, assessing both lab-scale and large-scale production. It identifies key chemical contributors to their environmental impact and cumulative energy demand (CED). PTFE has a lower CED in regions with renewable energy, while PVDF may be viable in areas reliant on non-renewable biomass. These insights can inform decision-makers in strategizing the implementation of TOEC processes for sustainable development.

Abstract Objective With an increasingly ageing population and osteoarthritis prevalence, the quantification of nociceptive signals responsible for painful movements and individual responses could lead to better treatment and monitoring solutions. Changes in electrodermal activity (EDA) can be detected via changes in skin conductance (SC) and measured using finger electrodes on a wearable sensor, providing objective information for increased physiological stress response. Results To provide EDA response preliminary data, this was recorded with healthy volunteers on an array of activities while receiving a noxious stimulus. This provides a defined scenario that can be utilised as protocol feasibility testing. Raw signal extraction, processing and statistical analysis was performed using mean SC values on all participant data. The application of the stimuli resulted in a significant average increase (p

Introduction Sleepiness while driving is potentially fatal, and it is recommended that a driver who starts to feel tired should stop and have a rest. However, some may use various countermeasures to try to stay alert. We devised a questionnaire that assessed various potential coping strategies that might be used against fatigue and compared them between obstructive sleep apnoea syndrome (OSAS) patients and controls and with sleepiness in general (Epworth Sleepiness Scale (ESS)), specifically while driving (Driving Sleepiness Scale (DSS)) and driving incidents. Methods 119 untreated OSAS patients (male 82%, body mass index (BMI) 37±8 kg·m−2, ESS 14±5, DSS 3±2, oxygen desaturation index (ODI) 39±15) and 105 controls (male 70%, BMI 28±6 kg·m−2, ESS 4±3, DSS 7±6) matched for age and driving experience were recruited. All completed a questionnaire relating to their experience over the last year, which included sleepiness in general, sleepiness specifically while driving, 10 questions about various coping strategies they might adopt in order to avoid sleepiness and their history of incidents while driving. Results As compared to controls, nearly a third of OSAS patients (29.4%) used more than three coping strategies “frequently”. OSAS patients who used more than three such strategies had worse ESS (17±4 versus 12±5, p

BackgroundThere is increasing evidence that co-design can lead to more engaging, acceptable, relevant, feasible, and even effective interventions. However, no guidance is provided on the specific designs and associated methods or methodologies involved in the process. We propose the development of the Preferred Components for Co-design in Research (PRECISE) guideline to enhance the consistency, transparency, and quality of reporting co-design studies used to develop complex health interventions. ObjectiveThe aim is to develop the first iteration of the PRECISE guideline. The purpose of the PRECISE guideline is to improve the consistency, transparency, and quality of reporting on studies that use co-design to develop complex health interventions. MethodsThe aim will be achieved by addressing the following objectives: to review and synthesize the literature on the models, theories, and frameworks used in the co-design of complex health interventions to identify their common elements (components, values or principles, associated methods and methodologies, and outcomes); and by using the results of the scoping review, prioritize the co-design components, values or principles, associated methods and methodologies, and outcomes to be included in the PRECISE guideline. ResultsThe project has been funded by the Canadian Institutes of Health Research. ConclusionsThe collective results of this project will lead to a ready-to-implement PRECISE guideline that outlines a minimum set of items to include when reporting the co-design of complex health interventions. The PRECISE guideline will improve the consistency, transparency, and quality of reports of studies. Additionally, it will include guidance on how to enact or enable the values or principles of co-design for meaningful and collaborative solutions (interventions). PRECISE might also be used by peer reviewers and editors to improve the review of manuscripts involving co-design. Ultimately, the PRECISE guideline will facilitate more efficient use of new results about complex health intervention development and bring better returns on research investments. International Registered Report Identifier (IRRID)PRR1-10.2196/50463

Cationic methylene blue (MB) and anionic orange G (OG) dyes were adsorbed using the first-ever synthesized nanocomposite of MXene-AgMOF. At 200 mg/L and 0.01 g, GO-AgMOF, MXene-AgMOF, and AgMOF were able to adsorb 99.9%, 99.0%, and 98.0% of cationic MB dye, respectively, from water. The nanocomposites were characterized both before and after adsorption using different characterization techniques. These nanocomposites show promise as cationic contaminant adsorbents, with an adsorption capacity of 399.9 mg/g for GO-AgMOF. Also, the enhanced adsorption capacity of AgMOF for anionic and cationic dyes suggests its potential use in environmental remediation when combined with MXene.

Objective Noninvasive ventilation (NIV) improves survival and quality of life in motor neuron disease (MND), but many patients fail to receive effective ventilation. This study aimed to map the respiratory clinical care for MND patients at a service and individual healthcare professional (HCP) level to understand where attention may be needed to ensure all patients receive optimal care. Methods Two online surveys of HCPs working with MND patients in the UK were conducted. Survey 1 targeted HCPs providing specialist MND care. Survey 2 targeted HCPs working in respiratory/ventilation services and community teams. Data were analysed using descriptive and inferential statistics. Results Responses from 55 HCPs providing specialist MND care who worked at 21 MND care centres and networks and 13 Scotland Health Boards were analysed from Survey 1. Responses from 85 HCPs from respiratory/ventilation services and 73 HCPs from community teams, representing 97 services, were analysed from Survey 2. Significant differences in practice were identified at each stage of the respiratory care pathway as well as evidence of the need for improvement. This included when patients were referred to respiratory services, the time taken waiting to commence NIV, the availability of sufficient NIV equipment and provision of services, particularly out of hours. Conclusion We have highlighted significant disparity in MND respiratory care practices. Increased awareness of the factors that influence NIV success and the performance of individuals and services is important for optimal practice.

Abstract Eurasian beaver (Castor fiber) were nearly hunted to extinction but have recovered to occupy much of their former range. Beaver were extirpated from Great Britain c. 400 years ago but have recently been reintroduced. The River Otter catchment, Devon was the site of the first licensed wild release of beavers in England. With further releases being considered, there is a need to better understand population dynamics of this native, keystone species to inform conservation and management. Field signs were surveyed from 2015 to 2021. A semi‐automated territory detection method was adopted to estimate territory counts. A spatially explicit model was developed to estimate the ecological territory capacity of the catchment. Future territory expansion was modelled using logistic growth curves; initial growth rate was estimated from observed territory counts and the estimated territory capacity range was used to define the limiting value of the growth curve. Beaver territory removal was simulated, across a range of management intensities and start times, to determine potential impacts of translocation or lethal control upon population dynamics. Territory numbers increased from four to 18, inclusive of four additionally released individuals, during study period. In the absence of population management, the territory capacity of the catchment was estimated to range between 120 and 183; this may be reached between 2028 and 2057. Simulated territory removal, where territories were removed at a fixed rate from the sum of the estimated total population and the population increase for that year, demonstrated large uncertainties in predicted population responses. Simulations with territory removals >3/year all predicted potential population collapse. This finding emphasizes the need for caution when considering population management strategies; translocation of animals out of the catchment or culling should be considered only when populations are established and all alternatives have been considered. These results provide critical information for the expected rate and magnitude of beaver population change in the River Otter catchment. The methods provide a reproducible and generalizable approach for understanding beaver population change, which can inform policy on the reintroduction of beavers and the potential timing and intensity of future beaver population management.

Background: No periprocedural metric has demonstrated improved cardiac resynchronization therapy (CRT) outcomes in a multicenter setting. Objective: We sought to determine if left ventricular (LV) lead placement targeted to the coronary sinus (CS) branch generating the best acute hemodynamic response (AHR) results in improved outcomes at 6 months. Methods: In this multicenter randomized controlled trial, patients were randomized to guided CRT or conventional CRT. Patients in the guided arm had LV dP/dtmax measured during biventricular (BIV) pacing. Target CS branches were identified and the final LV lead position was the branch with the best AHR and acceptable threshold values. The primary endpoint was the proportion of patients with a reduction in LV end-systolic volume (LVESV) of ≥15% at 6 months. Results: A total of 281 patients were recruited across 12 centers. Mean age was 70.8 ± 10.9 years and 54% had ischemic etiology. Seventy-three percent of patients in the guided arm demonstrated a reduction in LVESV of ≥15% at 6 months vs 60% in the conventional arm (P = .02). Patients with AHR ≥ 10% were more likely to demonstrate a reduction of ESV ≥ 15% (84% of patients with an AHR ≥10% vs 28% with an AHR

Background: Cardiac resynchronization therapy (CRT) produces acute changes in electric resynchronization that can be measured noninvasively with electrocardiographic body surface mapping (ECGi). The relation between baseline acute electrophysiology metrics and their manipulation with CRT and reverse remodeling is unclear. Objective: To test (ECGi) derived parameters of electrical activation as predictors of volumetric response to CRT. Methods: ECGi was performed in 21 patients directly following CRT implant. Activation parameters (left ventricular total activation time [LVtat], global biventricular total activation time [VVtat], global left/right ventricular electrical synchrony [VVsync], and global left ventricular dispersion of activation times [LVdisp]) were measured at baseline and following echocardiographically optimized CRT. Remodeling response (>15% reduction left ventricular end-systolic volume) was assessed 6 months post CRT. Results: Patients were aged 68.9 ± 12.1 years, 81% were male, and 57% were ischemic. Baseline measures of dyssynchrony were more pronounced in left bundle branch block (LBBB) vs non-LBBB. ECGi demonstrated a trend of greater interventricular dyssynchrony between responders and nonresponders that did not reach statistical significance (VVsync: -45.7 ± 22.4 ms vs -25.1 ± 29.3 ms, P = .227). Remaining activation parameters were similar between responders and nonresponders (VVtat 101 ± 22.0 ms vs 98.9 ± 23.4 ms, P = .838; LVtat 86.4 ± 17.1 ms vs 85.1 ± 27.7 ms, P = .904; LVdisp 28.2 ± 6.3 ms vs 27.0 ± 8.7 ms, P = .726). In volumetric responders activation parameters were significantly improved with CRT compared to nonresponders: VV sync (-45.67 ± 22.41 ms vs 2.33±18.87 ms, P = .001), VVtat (101 ± 22.04 ms vs 71 ± 14.01 ms, P = .002), LVtat (86.44 ± 17.15 ms vs 67.67 ± 11.31 ms, P = .006), and LVdisp (28.22 ± 6.3 ms vs 21.56 ± 4.45 ms, P = .008). Conclusion: Baseline ECGi activation times did not predict CRT volumetric response. Volumetric responders exhibited significant improvements in ECGi-derived metrics with CRT. ECGi does not select CRT candidates but may be a useful adjunct to guide left ventricle lead implants and to perform postimplant CRT optimization.

MXenes have significantly impacted materials science and nanotechnology since their discovery in 2011. Theoretical calculations have predicted more than 100 possible compositions of MXenes and lab-scale fabrication of more than 40 MXene structures has been reported to date. The unique characteristics of MXenes have made them an ideal fit for a wide variety of applications, including energy storage, environmental, electronics, communications, gas and liquid separations and adsorption, biomedical, and optoelectronics. MXene attracted many researchers, and as a result, publication trends on MXene have grown exponentially in recent years. By 2021, MXenes have already shown promise in several research areas, including energy storage devices, electromagnetic interference shielding, nanocomposites, and hybrid materials. In parallel, new applications are emerging where MXenes outperform other nanomaterials, such as in tribology. MXene compositions are also being expanded rapidly. Here, we briefly overview the history, properties, trends, and application of MXenes to better understand their potentials and familiarize new audiences with this 2D material family.

This case studied the treatment of cheese whey wastewater (CWW) from the Kalleh® dairy industry using a membrane bioreactor (MBR). The coagulation process and activated sludge sections eliminated 26% and 75% of organic matter from CWW with chemical oxygen demand (COD) of 10,000 mg/l, respectively. In the MBR, metal-organic framework (MOF)-modified PSf membranes enhanced the whole system's performance. The flux of water and CWW increased from 157 and 28 (L/m2.h) to 350 and 60 (L/m2.h), respectively, by only 2 wt% MOF concentration. The reduction of COD in the overall system was around 98.8%.

Background: Children and young people (CYP) with chronic rheumatic conditions; Juvenile Idiopathic Arthritis, Juvenile Systemic Lupus Erythematosus, Juvenile Dermatomyositis and Juvenile Vasculitis, treated with steroids, have low bone density, increased fracture risk and are likely to have suboptimal peak bone mass. There is currently no evidence base for the management of steroid-induced bone loss in children with rheumatic diseases. Methods: We undertook a multi-centre double dummy double-blind randomised placebo controlled trial to investigate whether the bisphosphonate risedronate was superior to alfacalcidol or calcium and vitamin D supplementation in the prevention and treatment of steroid-induced osteopaenia in these children. Patients were stratified and randomised in a 1:1 ratio, into: placebo; alfacalcidol; risedronate. The primary outcome was the change in lumbar spine bone mineral density z score (LSaBMDz) measured by dual energy x-ray absorptiometry at one year. Secondary outcome was fracture rate. Results: Two hundred and seventeen patients were recruited to the study. Seventy seven placebo, 71 alfacalcidol, and 69 risedronate. Highly statistically significant differences were observed in the change in LSaBMDz between the placebo and risedronate groups; 0.274, 95% CI (0.061, 0.487) (p

Produced water (PW) generation has been increasing recently due to the expansion of fossil fuel extraction and the aging of oil wells worldwide, especially in the United States. The adverse health risks, seismicity, and environmental impacts associated with PW have become a challenging concern. Therefore, there is increased demand for improved PW treatment and reuse management options. There are multiple methods for treating PW; this article focuses on treatment through membrane filtration. Moreover, this mini review aims to summarize statistics on PW abundance and trends in PW generation over time, to briefly call attention to health-related issues, highlight some treatment challenges, and mention the potential purposes for reuse with an emphasis on the United States, the largest generator of PW worldwide.

Objective: This UK-wide OATech Network + consensus study utilised a Delphi approach to discern levels of awareness across an expert panel regarding the role of existing and novel technologies in osteoarthritis research. To direct future cross-disciplinary research it aimed to identify which could be adopted to subcategorise patients with osteoarthritis (OA). Design: An online questionnaire was formulated based on technologies which might aid OA research and subcategorisation. During a two-day face-to-face meeting concordance of expert opinion was established with surveys (23 questions) before, during and at the end of the meeting (Rounds 1, 2 and 3, respectively). Experts spoke on current evidence for imaging, genomics, epigenomics, proteomics, metabolomics, biomarkers, activity monitoring, clinical engineering and machine learning relating to subcategorisation. For each round of voting, ≥80% votes led to consensus and ≤20% to exclusion of a statement. Results: Panel members were unanimous that a combination of novel technological advances have potential to improve OA diagnostics and treatment through subcategorisation, agreeing in Rounds 1 and 2 that epigenetics, genetics, MRI, proteomics, wet biomarkers and machine learning could aid subcategorisation. Expert presentations changed participants’ opinions on the value of metabolomics, activity monitoring and clinical engineering, all reaching consensus in Round 2. X-rays lost consensus between Rounds 1 and 2; clinical X-rays reached consensus in Round 3. Conclusion: Consensus identified that 9 of the 11 technologies should be targeted towards OA subcategorisation to address existing OA research technology and knowledge gaps. These novel, rapidly evolving technologies are recommended as a focus for emergent, cross-disciplinary osteoarthritis research programmes.

Disparate diagnostic categories from the Diagnostic and Statistical Manual of Mental Disorders (DSM), including generalized anxiety disorder, major depressive disorder and post-traumatic stress disorder, share common behavioral and phenomenological dysfunctions. While high levels of comorbidity and common features across these disorders suggest shared mechanisms, past research in psychopathology has largely proceeded based on the syndromal taxonomy established by the DSM rather than on a biologically-informed framework of neural, cognitive and behavioral dysfunctions. In line with the National Institute of Mental Health’s Research Domain Criteria (RDoC) framework, we present a Human Connectome Study Related to Human Disease that is intentionally designed to generate and test novel, biologically-motivated dimensions of psychopathology. The Dimensional Connectomics of Anxious Misery study is collecting neuroimaging, cognitive and behavioral data from a heterogeneous population of adults with varying degrees of depression, anxiety and trauma, as well as a set of healthy comparators (to date, n = 97 and n = 24, respectively). This sample constitutes a dataset uniquely situated to elucidate relationships between brain circuitry and dysfunctions of the Negative Valence construct of the RDoC framework. We present a comprehensive overview of the eligibility criteria, clinical procedures and neuroimaging methods of our project. After describing our protocol, we present group-level activation maps from task fMRI data and independent components maps from resting state data. Finally, using quantitative measures of neuroimaging data quality, we demonstrate excellent data quality relative to a subset of the Human Connectome Project of Young Adults (n = 97), as well as comparable profiles of cortical thickness from T1-weighted imaging and generalized fractional anisotropy from diffusion weighted imaging. This manuscript presents results from the first 121 participants of our full target 250 participant dataset, timed with the release of this data to the National Institute of Mental Health Data Archive in fall 2020, with the remaining half of the dataset to be released in 2021.

The effect of surface roughness on convective heat transfer and pressure drop is assessed for a simplified pre-cooler system for application in high-speed airbreathing engines. The pre-cooler system comprises of four circular cylinders in cross-flow inside a square duct. The cylindrical wires of 1 mm diameter were textured using an economical method of surface treatment, via sand-blasting with 100 µm (diameter) particles. Experiments were conducted at Reynolds numbers ranging from 600 to 8000. The Nusselt number was observed to decrease for the rough wires, indicating decreased convective heat transfer rates. Moreover, roughness was found to have no appreciable effect on pressure drop for these conditions. This is attributed to a fully developed turbulent duct flow upstream of the wires, which was confirmed by observing the Nusselt-Reynolds number correlation for treated and untreated cases. Surface roughness strategy in pre-coolers for an ascent trajectory of a high-speed airbreathing engine is also discussed. Keywords: Convective heat transfer, Surface roughness, Heat exchangers, High-speed airbreathing engines

Gemtuzumab ozogamicin (GO) is an anti-CD33 antibody-drug conjugate for the treatment of acute myeloid leukemia (AML). Although GO shows a narrow therapeutic window in early clinical studies, recent reports detailing a modified dosing regimen of GO can be safely combined with induction chemotherapy, and the combination provides significant survival benefits in AML patients. Here we tested whether the survival benefits seen with the combination arise from the enhanced reduction of chemoresidual disease and leukemic initiating cells (LICs). Herein, we use cell line and patient-derived xenograft (PDX) AML models to evaluate the combination of GO with daunorubicin and cytarabine (DA) induction chemotherapy on AML blast growth and animal survival. DA chemotherapy and GO as separate treatments reduced AML burden but left significant chemoresidual disease in multiple AML models. The combination of GO and DA chemotherapy eliminated nearly all AML burden and extended overall survival. In two small subsets of AML models, chemoresidual disease following DA chemotherapy displayed hallmark markers of leukemic LICs (CLL1 and CD34). In vivo, the two chemoresistant subpopulations (CLL1+/CD117− and CD34+/CD38+) showed higher ability to self-renewal than their counterpart subpopulations, respectively. CD33 was coexpressed in these functional LIC subpopulations. We demonstrate that the GO and DA induction chemotherapy combination more effectively eliminates LICs in AML PDX models than either single agent alone. These data suggest that the survival benefit seen by the combination of GO and induction chemotherapy, nonclinically and clinically, may be attributed to the enhanced reduction of LICs.

Botulinum neurotoxins (BoNTs) are notorious toxins and powerful agents and can be lethal, causing botulism, but they are also widely used as therapeutics, particularly to treat neuromuscular disorders. As of today, the commercial BoNT treatments available are from native A or B serotypes. Serotype F has shown efficacy in a clinical trial but has scarcely been used, most likely due to its medium duration of effect. Previously, the uniqueness of the light chain of the F7 subtype was identified and reported, showing an extended interaction with its substrates, VAMPs 1, 2 and 3, and a superior catalytic activity compared to other BoNT/F subtypes. In order to more extensively study the properties of this neurotoxin, we engineered a modified F7 chimera, mrBoNT/F7-1, in which all the regions of the neurotoxin were identical to BoNT/F7 except the activation loop, which was the activation loop from BoNT/F1. Use of the activation loop from BoNT/F1 allowed easier post-translational proteolytic activation of the recombinant protein without otherwise affecting its properties. mrBoNT/F7-1 was expressed, purified and then tested in a suite of in vitro and in vivo assays. mrBoNT/F7-1 was active and showed enhanced potency in comparison to both native and recombinant BoNT/F1. Additionally, the safety profile remained comparable to BoNT/F1 despite the increased potency. This new modified recombinant toxin F7 could be further exploited to develop unique therapeutics to address unmet medical needs.

Botulinum neurotoxin (BoNT) serotype A inhibits neurotransmitter release by cleaving SNAP-25 and represents an established pharmaceutical for treating medical conditions caused by hyperactivity of cholinergic nerves. Oversecretion from non-neuronal cells is often also the cause of diseases. Notably, excessive release of inflammatory messengers is thought to contribute to diseases such as chronic obstructive pulmonary disease, asthma, diabetes etc. The expansion of its application to these medical conditions is prevented because the major non-neuronal SNAP-25 isoform responsible for exocytosis, SNAP-23, is, in humans, virtually resistant to BoNT/A. Based on previous structural data and mutagenesis studies of SNAP-23 we optimized substrate binding pockets of the enzymatic domain for interaction with SNAP-23. Systematic mutagenesis and rational design yielded the mutations E148Y, K166F, S254A, and G305D, each of which individually increased the activity of LC/A against SNAP-23 between 3- to 23-fold. The assembled quadruple mutant showed approximately 2000-fold increased catalytic activity against human SNAP-23 in in vitro cleavage assays. A comparable increase in activity was recorded for the full-length BoNT/A quadruple mutant tested in cultivated primary neurons transduced with a fluorescently tagged-SNAP-23 encoding gene. Equipped with a suitable targeting domain this quadruple mutant promises to complete successfully tests in cells of the immune system.