MEF2C regulates NK cell effector functions through control of lipid metabolism.

Natural killer (NK) cells are a critical first line of defense against viral infection. Rare mutations in a small subset of transcription factors can result in decreased NK cell numbers and function in humans, with an associated increased susceptibility to viral infection. However, our understanding of the specific transcription factors governing mature human NK cell function is limited. Here we use a non-viral CRISPR-Cas9 knockout screen targeting genes encoding 31 transcription factors differentially expressed during human NK cell development. We identify myocyte enhancer factor 2C (MEF2C) as a master regulator of human NK cell functionality ex vivo. MEF2C-haploinsufficient patients and mice displayed defects in NK cell development and effector function, with an increased susceptibility to viral infection. Mechanistically, MEF2C was required for an interleukin (IL)-2- and IL-15-mediated increase in lipid content through regulation of sterol regulatory element-binding protein (SREBP) pathways. Supplementation with oleic acid restored MEF2C-deficient and MEF2C-haploinsufficient patient NK cell cytotoxic function. Therefore, MEF2C is a critical orchestrator of NK cell antiviral immunity by regulating SREBP-mediated lipid metabolism.

Single-center outcomes of Onyx Frontier™ and Resolute Onyx™ drug-eluting balloon-mounted stents for rescue stenting for acute large vessel occlusion.

BACKGROUND: In cases where mechanical thrombectomy (MT) fails, rescue stenting may be necessary to achieve reperfusion; however, the lack of standardized techniques or devices poses a challenge. This series aims to present our early experience with the Onyx Frontier™ and Resolute Onyx™ balloon-mounted drug-eluting stents for rescue stenting. METHODS: A retrospective chart review was performed of all patients who underwent rescue stenting, in the setting of failed MT, using Onyx Frontier™ or Resolute Onyx™ stents at a single institution. Technical details, procedural complications, and patient outcomes were recorded for each case. RESULTS: Twenty-two Onyx Frontier™ and Resolute Onyx™ stents were deployed in 18 patients undergoing rescue stenting. Stent locations included the middle cerebral artery (36.4%), internal carotid artery (18.2%), vertebral artery (22.7%), and basilar artery (22.7%). The average National Institutes of Health Stroke Scale score before MT was 13.8 (range 0-31). The median initial modified Rankin Scale (mRS) score was zero, while the median mRS score at follow-up was three. Successful reperfusion, as assessed by TICI scores, was achieved in 43.8% of patients for TICI 3, 43.8% for TICI 2C, and 12.5% for TICI 2B. Post-revascularization, 16.7% of patients experienced hemorrhage, of which one patient (5.6%) had symptomatic hemorrhage. CONCLUSIONS: Onyx Frontier™ and Resolute Onyx™ stents are well suited for rescue stenting in cases of failed MT. These balloon-mounted drug-eluting stents exhibit excellent navigability, rendering them appropriate for rescue revascularization procedures. Our findings demonstrate that these stents confer a high degree of technical success.

An energy tunable continuous 23S1 positronium beam.

We describe the experimental production of a beam of 23S1 positronium (Ps) atoms obtained from charge-exchange collisions between a positron beam and Xe held in a gas cell. The angular divergence of the emitted Ps beam was recorded using two position sensitive detectors located at different distances from the gas cell. The fraction of the Ps beam produced in the 23S1 level was measured via the change in the Ps count rate after driving the 23S1 → 23P2 transition with microwave radiation; with optimal experimental parameters, we estimate that up to 10% of the Ps beam is formed in the 23S1 state. The measured properties of the beam were used to evaluate the feasibility of using the system for precision spectroscopy of the n = 2 Ps fine structure using Ramsey interferometry.

Human retinal ganglion cell neurons generated by synchronous BMP inhibition and transcription factor mediated reprogramming.

In optic neuropathies, including glaucoma, retinal ganglion cells (RGCs) die. Cell transplantation and endogenous regeneration offer strategies for retinal repair, however, developmental programs required for this to succeed are incompletely understood. To address this, we explored cellular reprogramming with transcription factor (TF) regulators of RGC development which were integrated into human pluripotent stem cells (PSCs) as inducible gene cassettes. When the pioneer factor NEUROG2 was combined with RGC-expressed TFs (ATOH7, ISL1, and POU4F2) some conversion was observed and when pre-patterned by BMP inhibition, RGC-like induced neurons (RGC-iNs) were generated with high efficiency in just under a week. These exhibited transcriptional profiles that were reminiscent of RGCs and exhibited electrophysiological properties, including AMPA-mediated synaptic transmission. Additionally, we demonstrated that small molecule inhibitors of DLK/LZK and GCK-IV can block neuronal death in two pharmacological axon injury models. Combining developmental patterning with RGC-specific TFs thus provided valuable insight into strategies for cell replacement and neuroprotection.

Precision Microwave Spectroscopy of the Positronium 2

We report the results of a new measurement of the positronium 2 ^{3}S_{1}→2 ^{3}P_{2} (ν_{2}) interval. Using a modified experimental arrangement we have significantly reduced the effects of microwave reflections, which in previous experiments resulted in shifts and asymmetric line shapes. With the improved apparatus we obtain an experimental value of ν_{2}=8627.94±0.95 MHz, which is within 1.3σ of the theoretical value 8626.71±0.08 MHz.

The role of a photographic atlas in reducing unanticipated healthcare utilization following circumcision.

INTRODUCTION: Circumcision is a common procedure that can evoke caregiver anxiety in the postoperative period due to unfamiliarity with the healing process. To mitigate unnecessary healthcare utilization such as phone calls and unanticipated clinic or emergency department (ED) visits, photographic atlases have been developed to better prepare caregivers for the recovery process. The objective of our study is to further investigate the efficacy of a photographic atlas in its ability to decrease postoperative healthcare utilization using an increased sample size and extended study period compared to previous studies. MATERIALS AND METHODS: In this study, we compared a prospective intervention cohort of patients undergoing circumcision at our institution who received a photographic atlas during postoperative teaching to a retrospective cohort of patients who had not received it. Our primary outcome was unanticipated healthcare utilization, defined as postoperative telephone calls and unanticipated presentations to the urology clinic or ED. RESULTS: The retrospective no-atlas cohort included 105 patients, and the prospective intervention atlas cohort included 80 patients. Both groups were similar with respect to age (p = 0.47) and other demographics. There was no statistically significant difference in healthcare utilization between the no-atlas and atlas cohort. Specifically, we identified no difference in the number of phone calls to clinic staff (12 [11.4%] vs. 11 [13.8%], p = 0.64) or unanticipated postoperative clinic or ED visits (2 [1.9%] vs. 4 [5.0%], p = 0.41). DISCUSSION: The use of a photographic atlas as part of caregiver support for circumcision patients did not demonstrate a statistically significant reduction in either postoperative phone calls or clinic/ED visits. The decrease in absolute number of caregiver phone calls was minimal (12-11), with a small increase in follow-up presentations (2-4). The lack of significant change may be due to the already infrequent occurrence of these events following circumcision, as demonstrated by the no-atlas cohort. Other potential advantages of the atlas, such as improved caregiver confidence and satisfaction, may have been present, but were not measured in this study. CONCLUSIONS: Adding to the mixed results of previous studies, these findings do not support that photographic atlases decrease unanticipated healthcare utilization in children undergoing a circumcision. However, utilization was found to be low. Additionally, further studies are needed to determine other significant benefits of this form of education, such as improved caregiver confidence and satisfaction.

Endoscopically-Assisted Percutaneous Trigeminal Rhizotomy for Trigeminal Neuralgia: A Cadaveric Feasibility Study.

Objective Trigeminal neuralgia (TN) is a debilitating syndrome characterized by paroxysmal facial pain in one or more divisions of the trigeminal nerve. The etiology and treatment paradigms are still controversial. The endoscopically-assisted procedure has not yet been described in percutaneous procedures for TN. The aim of this study was to assess the utility and feasibility of endoscopic-assisted percutaneous approaches for trigeminal rhizotomy in TN. Methods This study comprised eight cadaveric sides heads that underwent an endoscopically assisted percutaneous approach using Hakanson's anterior puncture method for targeting the foramen ovale. Results V3 exiting the foramen ovale was easily visualized with the endoscope on all sides. While approaching the foramen ovale, distal branches of V3 such as the lingual and inferior alveolar nerves were first identified as they traveled between the medial and lateral pterygoid muscles. These branches were then traced proximally to the V3 trunk deep to the lateral pterygoid. Large arteries and veins were easily visualized and avoided in the trajectory to the foramen ovale. No gross injury to any neurovascular structure along the course of the needle insertion was identified. Conclusion We found that endoscopic-assisted percutaneous approach to the foramen ovale is feasible and allows for accurate canalization and anatomical identification of the precise location for rhizotomy under direct visualization. Such a procedure, after it is confirmed in patients, could offer a new technique for reducing unsuccessful canalization and could improve outcomes.

Ventral Spinal Cord Herniation Causing Spinal Intradural Hematoma and Subarachnoid Hemorrhage: A Case Report.

Ventral spinal cord herniation is a rare pathology, caused by a dural defect, that leads to progressive myelopathy. The true prevalence of ventral spinal cord herniation is unknown largely because of underdiagnosis due to its nonspecific symptoms. Though there are theories that attempt to describe how these dural defects are formed, the true causes of these defects are unknown. In this case report, we present a case of a 29-year-old female who had an idiopathic ventral spinal cord herniation causing an intradural hematoma and subarachnoid hemorrhage. This is the first reported case of spinal cord herniation causing hemorrhage.

Transradial access for anterior circulation deployment of pipeline embolization device: A case report, literature review, and technical note.

Common femoral artery (CFA) transfemoral access (TFA) has been the traditional route for neuroendovascular intervention with flow diversion including the pipeline embolization device (PED) for the treatment of wide-necked aneurysms. Successful deployment requires significant catheter support, thus making alternative access challenging. A 56-year-old-female presented with subarachnoid hemorrhage secondary to a large ruptured posterior communicating artery (PCOM) aneurysm as well as found to have an unruptured left superior cerebellar artery (SCA) aneurysm. Endovascular embolization of PCOM aneurysm via TFA was complicated by a right CFA pseudoaneurysm. The SCA aneurysm was treated 8 weeks later via left TFA with consequent development of a left CFA pseudoaneurysm. Contrasted magnetic resonance angiography revealed recurrence at the neck of the PCOM aneurysm at 4-month follow-up, treated via transradial access (TRA) PED flow diversion to avoid additional groin complications. Anatomic, procedural, and clinical considerations for TRA anterior circulation flow diversion using the PED are reviewed.

Prevention through design: insights from computational fluid dynamics modeling to predict exposure to ultrafine particles from 3D printing.

Fused filament fabrication (FFF) 3D printers are increasingly used in industrial, academic, military, and residential sectors, yet their emissions and associated user exposure scenarios are not fully described. Characterization of potential user exposure and environmental releases requires robust investigation. During operation, common FFF 3D printers emit varying amounts of ultrafine particles (UFPs) depending upon feedstock material and operation procedures. Volatile organic compounds associated with these emissions exhibit distinct odors; however, the UFP portion is largely imperceptible by humans. This investigation presents straightforward computational modeling as well as experimental validation to provide actionable insights for the proactive design of lower exposure spaces where 3D printers may be used. Specifically, data suggest that forced clean airflows may create lower exposure spaces, and that computational modeling might be employed to predict these spaces with reasonable accuracy to assist with room design. The configuration and positioning of room air ventilation diffusers may be a key factor in identifying lower exposure spaces. A workflow of measuring emissions during a printing process in an ANSI/CAN/UL 2904 environmental chamber was used to provide data for computational fluid dynamics (CFD) modeling of a 6 m2 room. Measurements of the particle concentrations in a Class 1000 clean room of identical geometry were found to pass the Hanna test for agreement between model and experimental data, validating the findings.

Inhibition of GCK-IV kinases dissociates cell death and axon regeneration in CNS neurons.

Axon injury is a hallmark of many neurodegenerative diseases, often resulting in neuronal cell death and functional impairment. Dual leucine zipper kinase (DLK) has emerged as a key mediator of this process. However, while DLK inhibition is robustly protective in a wide range of neurodegenerative disease models, it also inhibits axonal regeneration. Indeed, there are no genetic perturbations that are known to both improve long-term survival and promote regeneration. To identify such a neuroprotective target, we conducted a set of complementary high-throughput screens using a protein kinase inhibitor library in human stem cell-derived retinal ganglion cells (hRGCs). Overlapping compounds that promoted both neuroprotection and neurite outgrowth were bioinformatically deconvoluted to identify specific kinases that regulated neuronal death and axon regeneration. This work identified the role of germinal cell kinase four (GCK-IV) kinases in cell death and additionally revealed their unexpected activity in suppressing axon regeneration. Using an adeno-associated virus (AAV) approach, coupled with genome editing, we validated that GCK-IV kinase knockout improves neuronal survival, comparable to that of DLK knockout, while simultaneously promoting axon regeneration. Finally, we also found that GCK-IV kinase inhibition also prevented the attrition of RGCs in developing retinal organoid cultures without compromising axon outgrowth, addressing a major issue in the field of stem cell-derived retinas. Together, these results demonstrate a role for the GCK-IV kinases in dissociating the cell death and axonal outgrowth in neurons and their druggability provides for therapeutic options for neurodegenerative diseases.

Precision Microwave Spectroscopy of the Positronium n=2 Fine Structure.

We report a new measurement of the positronium (Ps) 2^{3}S_{1}→2^{3}P_{0} interval. Slow Ps atoms, optically excited to the radiatively metastable 2^{3}S_{1} level, flew through a microwave radiation field tuned to drive the transition to the short-lived 2^{3}P_{0} level, which was detected via the time spectrum of subsequent ground state Ps annihilation radiation. After accounting for Zeeman shifts we obtain a transition frequency ν_{0}=18501.02±0.61 MHz, which is not in agreement with the theoretical value of ν_{0}=18498.25±0.08 MHz.

Serotonin, norepinephrine, and acetylcholine differentially affect astrocytic potassium clearance to modulate somatosensory signaling in male mice.

Changes in extracellular potassium ([K+ ]e ) modulate neuronal networks via changes in membrane potential, voltage-gated channel activity, and alteration to transmission at the synapse. Given the limited extracellular space in the central nervous system, potassium clearance is crucial. As activity-induced potassium transients are rapidly managed by astrocytic Kir4.1 and astrocyte-specific Na+ /K+ -ATPase, any neurotransmitter/neuromodulator that can regulate their function may have indirect influence on network activity. Neuromodulators differentially affect cortical/thalamic networks to align sensory processing with differing behavioral states. Given serotonin (5HT), norepinephrine (NE), and acetylcholine (ACh) differentially affect spike frequency adaptation and signal fidelity ("signal-to-noise") in somatosensory cortex, we hypothesize that [K+ ]e may be differentially regulated by the different neuromodulators to exert their individual effects on network function. This study aimed to compare effects of individually applied 5HT, NE, and ACh on regulating [K+ ]e in connection to effects on cortical-evoked response amplitude and adaptation in male mice. Using extracellular field and K+ ion-selective recordings of somatosensory stimulation, we found that differential effects of 5HT, NE, and ACh on [K+ ]e regulation mirrored differential effects on amplitude and adaptation. 5HT effects on transient K+ recovery, adaptation, and field post-synaptic potential amplitude were disrupted by barium (200 µM), whereas NE and ACh effects were disrupted by ouabain (1 µM) or iodoacetate (100 µM). Considering the impact [K+ ]e can have on many network functions; it seems highly efficient that neuromodulators regulate [K+ ]e to exert their many effects. This study provides functional significance for astrocyte-mediated buffering of [K+ ]e in neuromodulator-mediated shaping of cortical network activity.

Beyond Simple Differences: Moderators of Gender Differences in Heteronormativity.

Heteronormativity is a culturally embedded system defining acceptable identities and behaviors in romantic relationships. Recent literature has suggested that heteronormativity is strongly associated with political attitudes, personality characteristics such as tolerance of ambiguity, and key demographic variables such as gender and sexual orientation. However, interactions between gender and other factors have not been thoroughly explored as predictors of heteronormative attitude and beliefs. The current study (N = 306) expands the understanding of predictors of heteronormativity, particularly as they interact with gender. In the current study, higher education was associated with lower heteronormativity among women, but not among men, and expressiveness was associated with lower heteronormativity among men, but not among women. Finally, a 3-way interaction showed that the relationship between education and heteronormativity for men (but not women) depended on trait expressiveness. Future studies are recommended, including observation of changes in heteronormativity over time and exploration of mechanisms that reinforce or reduce heteronormativity.

Electrostatically Guided Rydberg Positronium.

We report experiments in which positronium (Ps) atoms were guided using inhomogeneous electric fields. Ps atoms in Rydberg-Stark states with principal quantum number n=10 and electric dipole moments up to 610 D were prepared via two-color two-photon optical excitation in the presence of a 670 V cm^{-1} electric field. The Ps atoms were created at the entrance of a 0.4 m long electrostatic quadrupole guide, and were detected at the end of the guide via annihilation gamma radiation. When the lasers were tuned to excite low-field-seeking Stark states, a fivefold increase in the number of atoms reaching the end of the guide was observed, whereas no signal was detected when high-field-seeking states were produced. The data are consistent with the calculated geometrical guide acceptance.

Controlling Positronium Annihilation with Electric Fields.

We show that the annihilation dynamics of excited positronium (Ps) atoms can be controlled using parallel electric and magnetic fields. To achieve this, Ps atoms were optically excited to n=2 sublevels in fields that were adjusted to control the amount of short-lived and long-lived character of the resulting mixed states. Inclusion of the former offers a practical approach to detection via annihilation radiation, whereas the increased lifetimes due to the latter can be exploited to optimize resonance-enhanced two-photon excitation processes (e.g., 1^{3}S→2^{3}P→nS/nD), either by minimizing losses through intermediate state decay, or by making it possible to separate the excitation laser pulses in time. In addition, photoexcitation of mixed states with a 2^{3}S_{1} component represents an efficient route to producing long-lived pure 2^{3}S_{1} atoms via single-photon excitation.

A trap-based pulsed positron beam optimised for positronium laser spectroscopy.

We describe a pulsed positron beam that is optimised for positronium (Ps) laser-spectroscopy experiments. The system is based on a two-stage Surko-type buffer gas trap that produces 4 ns wide pulses containing up to 5 × 10(5) positrons at a rate of 0.5-10 Hz. By implanting positrons from the trap into a suitable target material, a dilute positronium gas with an initial density of the order of 10(7) cm(-3) is created in vacuum. This is then probed with pulsed (ns) laser systems, where various Ps-laser interactions have been observed via changes in Ps annihilation rates using a fast gamma ray detector. We demonstrate the capabilities of the apparatus and detection methodology via the observation of Rydberg positronium atoms with principal quantum numbers ranging from 11 to 22 and the Stark broadening of the n = 2 → 11 transition in electric fields.

Selective production of Rydberg-stark states of positronium.

Rydberg positronium (Ps) atoms have been prepared in selected Stark states via two-step (1s→2p→nd/ns) optical excitation. Two methods have been used to achieve Stark-state selection: a field ionization filter that transmits the outermost states with positive Stark shifts, and state-selected photoexcitation in a strong electric field. The former is demonstrated for n=17 and 18 while the latter is performed for n=11 in a homogeneous electric field of 1.9 kV/cm. The observed spectral intensities and their dependence on the polarization of the laser radiation are in agreement with calculations that include the perturbations of the intermediate n=2 manifold. Our results pave the way for the generation of Rydberg Ps atoms with large electric dipole moments that are required for the realization of schemes to control their motion using inhomogeneous electric fields, an essential feature of some proposed Ps free-fall measurements requiring focused beams of long-lived atoms.