CRISPR genome engineering has become a powerful tool to functionally investigate the complex mechanisms of immune system regulation. While decades of work have aimed to genetically reprogram innate immunity, the utility of current approaches is restricted by poor knockout efficiencies or limited specificity for mature cell lineages in vivo. Here, we describe an optimized strategy for non-viral CRISPR-Cas9 ribonucleoprotein (cRNP) genomic editing of mature primary mouse innate lymphocyte cells (ILCs) and myeloid lineage cells that results in an almost complete loss of single or double target gene expression from a single electroporation. Furthermore, we describe in vivo adoptive transfer mouse models that can be utilized to screen for gene function during viral infection using cRNP-edited naive natural killer (NK) cells and bone-marrow-derived conventional dendritic cell precursors (cDCPs). This resource will enhance target gene discovery and offer a specific and simplified approach to gene editing in the mouse innate immune system.The influence of 3D microenvironments on apoptosis susceptibility remains poorly understood. Here, we studied the susceptibility of cancer cell spheroids, grown to the size of micrometastases, to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). Interestingly, pronounced, spatially coordinated response heterogeneities manifest within spheroidal microenvironments In spheroids grown from genetically identical cells, TRAIL-resistant subpopulations enclose, and protect TRAIL-hypersensitive cells, thereby increasing overall treatment resistance. TRAIL-resistant layers form at the interface of proliferating and quiescent cells and lack both TRAILR1 and TRAILR2 protein expression. In contrast, oxygen, and nutrient deprivation promote high amounts of TRAILR2 expression in TRAIL-hypersensitive cells in inner spheroid layers. COX-II inhibitor celecoxib further enhanced TRAILR2 expression in spheroids, likely resulting from increased ER stress, and thereby re-sensitized TRAIL-resistant cell layers to treatment. Our analyses explain how TRAIL response heterogeneities manifest within well-defined multicellular environments, and how spatial barriers of TRAIL resistance can be minimized and eliminated.A 77 year old gentleman, normally fit and well, was admitted with acute confusion. Calpeptin in vitro On admission GCS was 14/15, vital signs were within the normal limits and bilateral crepitation at the lung base. Head CT scan normal. CXR showed some air space opacification. Investigations revealed hyponatraemia, raised CRP and positive for COVID-19. Treated with antibiotics and intravenous saline, sodium returned to normal. Delirium remained unchanged four weeks post incidence. Neurological manifestations were documented in patients with COVID-19, however no report has shown delirium as a primary manifestation. This case illustrates acute confusion may be the only presenting symptom of COVID-19 without overt lung disease.Introduction The Neutrophil-to-Lymphocyte Ratio (NLR) has been investigated as an independent predictive marker for clinical outcomes in vascular diseases. This study aimed to investigate the peri-interventional behavior of NLR in patients with ruptured and unruptured intracranial aneurysms (IAs). Methods 117 patients with IAs, who were treated at our department and had available complete data, were retrospectively identified during a ten-year period. Routine laboratory parameters, including the neutrophil and lymphocytes counts, were evaluated before and after treatment. Results The baseline NLR showed significant differences between patients with ruptured and unruptured IAs (6.3 vs. 1.8; p less then 0.001). In patients with ruptured IAs, the baseline NLR decreased significantly during the follow-up visits, while in unruptured IAs, the NLR remained low. Furthermore, higher baseline NLR values could also be observed in patients with ruptured IAs and fatal outcome than in surviving patients (8.0 vs. 5.4; p=0.220). In patients with poor functional outcome, defined as modified Rankin Score ≥3, the NLR was significantly higher before treatment (p=0.047), at day 10 (p=0.025) and one month after treatment (p=0.001). Conclusions The peri-interventional NLR was significantly different between patients with ruptured and unruptured IAs. In patients with ruptured IAs, elevated baseline NLR levels were associated with poor postoperative functional outcomes and decreased postoperatively, implying the potential prognostic value of NLR in patients with IAs.Objective To adapt the Reaching Performance Scale for Stroke (RPSS) for the Wolf Motor Function Test (WMFT) “Lift Can” (Can) and “Hand to Box” (Box) items. Design Retrospective analysis of video-recorded WMFT assessment performed by three raters on two occasions; SETTING N/A; PARTICIPANTS 29 participants with mild to moderate upper extremity impairment, less then 3 months post stroke INTERVENTIONS N/A MAIN OUTCOME MEASURES Inter- and intra-rater agreement, concurrent validity of Wolf Motor Function Test-Reaching Performance Scale for Stroke (WMFT-RPSS) RESULTS Mean (SD) inter-rater Gwet’s agreement coefficient (AC2) was 0.61 (0.05) for Can WMFT-RPSS and 0.56 (0.03) for Box. Mean (SD) intra-rater AC2 for Can was 0.63 (0.05) and 0.70 (0.04) for Box. WMFT-RPSS Can and Box scores correlated with log mean WMFT time (C -0.73, B -0.48), Functional Ability Scale (C 0.87, B 0.62), Upper Extremity Fugl-Meyer Motor Score (C 0.69, B 0.51), and item movement rate (C 0.74, B 0.71), (all, p less then 0.05). Mean (SD) WMFT-RPSS score across the 29 participants was 12.7 (3.5) for Can (max score = 19) and 11.4 (3.0) for Box (max = 16). Conclusions WMFT-RPSS demonstrated moderate intra-rater and weak-to-moderate inter-rater agreement for individuals with mild-moderate impairment. For construct validity, Can and Box WMFT-RPSS were significantly correlated with four standardized measures. Average WMFT-RPSS scores revealed that some participants may have relied on compensatory movements to complete the task—a revelation not discernable from movement rate alone. The WMFT-RPSS is potentially useful as a valid and reliable tool to examine longitudinal changes in movement quality after stroke.