Standardized Temporary Atrial Epicardial Wire Locations Lead to Enhanced Atrial Signal Identification.

Background: Arrhythmias, common after pediatric cardiac surgery, are associated with increased morbidity and mortality. Atrial epicardial wires (AEW) improve diagnostic accuracy but have variable pacing and sensing properties based on their location. Even so, there are no longitudinal prospective pediatric studies examining ideal placement of AEW. Methods: This multicenter study compared atrial amplitudes, pacing sensitivities and thresholds via AEW placed at Bachmann's Bundle (BB) and the interatrial groove near the right pulmonary veins (IGRPV) versus the surgeons' standard locations. An AtriAmp system was used to obtain an atrial ECG to calculate atrial and ventricular amplitude from atrial electrograms on the bedside monitor. Sensitivities and thresholds via a temporary pacemaker were documented. ANOVA tests with repeated measures and post-hoc pairwise comparisons were performed to compare variables within the first 24-h postoperative hours. Mixed effects linear regression models were employed to examine daily trends. Results: In the first 24-h following cardiac surgery, AEW at BB and IGRPV showed significantly larger atrial amplitudes than the surgeons' standard locations. In addition, there was a negative trend in atrial ECG amplitude in all AEW from postoperative days 0 to 1; however, subsequent days showed a positive mean change in atrial amplitude with largest increase seen at BB. Atrial sensing as measured by the temporary pacemaker had statistically greater atrial signal amplitude from the BB-IGRPV set in both polarities (ie, with the BB as the - or + electrode pair) as compared to the surgeons' standard locations. No difference in atrial thresholds (mA) were noted in the immediate postoperative period or over time, with a relatively low atrial threshold at all sites. Conclusion: Standardization of AEW at Bachmann's Bundle can yield largest atrial amplitudes by atrial ECG and highest atrial sensing parameters without compromising atrial thresholds.

Preoperative Depression Screening in Patients with Distal Radius Fractures: An Evaluation of Its Modifiability on Outcomes for Patients with Depressive Disorder.

Background It is unclear whether nonpharmacologic intervention for depressive disorder (DD) in the preoperative period can prevent postoperative complications in hand surgery patients. Questions/Purpose The aims were to evaluate whether psychotherapy visits/depression screenings within 90 days of open reduction and internal fixation (ORIF) for distal radius fractures (DRFs) were associated with lower rates of (1) medical complications and (2) health care utilization (emergency department [ED] visits and readmissions). Methods A retrospective analysis of an administrative claims database from 2010 to 2021 was performed. DD patients who underwent ORIF for DRF were 1:5 propensity score matched by comorbidities, including those who did ( n = 8,993) and did not ( n = 44,503) attend a psychotherapy visit/depression screening 90 days before surgery. Multivariate logistic regression models were constructed to compare the odds ratio (OR) of medical complications, ED visits, and readmissions within 90 days. The p -values less than 0.001 were significant. Results DD patients who did not attend a preoperative psychotherapy visit/depression screening experienced fivefold higher odds of total medical complications (25.66 vs. 5.27%; OR: 5.25, p < 0.0001), including surgical site infections (1.23 vs. 0.14%; OR: 8.71, p < 0.0001), deep wound infections (0.98 vs. 0.17%; OR: 6.00, p < 0.0001), and transfusions (1.64 vs. 0.22%; OR: 7.61, p < 0.0001). Those who did not attend a psychotherapy visit/depression screening experienced higher odds of ED utilizations (9.71 vs. 2.71%; OR: 3.87, p < 0.0001), however, no difference in readmissions (3.40 vs. 3.54%; OR: 0.96, p = 0.569). Conclusion Depression screening may be a helpful preoperative intervention to optimize patients with DD undergoing hand surgery to minimize postoperative complications and health care utilization. Level of Evidence Level III.

An allosteric cyclin E-CDK2 site mapped by paralog hopping with covalent probes.

More than half of the ~20,000 protein-encoding human genes have paralogs. Chemical proteomics has uncovered many electrophile-sensitive cysteines that are exclusive to subsets of paralogous proteins. Here we explore whether such covalent compound-cysteine interactions can be used to discover ligandable pockets in paralogs lacking the cysteine. Leveraging the covalent ligandability of C109 in the cyclin CCNE2, we substituted the corresponding residue in paralog CCNE1 to cysteine (N112C) and found through activity-based protein profiling that this mutant reacts stereoselectively and site-specifically with tryptoline acrylamides. We then converted the tryptoline acrylamide-CCNE1-N112C interaction into in vitro NanoBRET (bioluminescence resonance energy transfer) and in cellulo activity-based protein profiling assays capable of identifying compounds that reversibly inhibit both the N112C mutant and wild-type CCNE1:CDK2 (cyclin-dependent kinase 2) complexes. X-ray crystallography revealed a cryptic allosteric pocket at the CCNE1:CDK2 interface adjacent to N112 that binds the reversible inhibitors. Our findings, thus, show how electrophile-cysteine interactions mapped by chemical proteomics can extend the understanding of protein ligandability beyond covalent chemistry.

Coolants, organic acids, flavourings and other additives that facilitate inhalation of tobacco and nicotine products: implications for regulation.

To inform regulatory policy, this article summarises findings on inhalation facilitation from the ninth report of the WHO Study Group on Tobacco Product Regulation. Some additives counteract the harshness and bitterness of tobacco and nicotine product aerosols, making them easier to inhale. Additives that promote inhalability may perpetuate and increase the use of inhaled tobacco and nicotine products, especially by young people. Thus, as a class, additives that facilitate inhalation are an important regulatory target to prevent tobacco and nicotine product uptake. We defined inhalation facilitation as modifications to products during manufacturing that enhance the sensory experience and (potentially) behaviours associated with inhalation (eg, deeper puffs, faster inhalation, larger puff volume, shorter intervals in between puffs and use episodes). Evidence review showed that: (a) menthol and synthetic coolants decrease irritation caused by aerosol constituents by activating sensory perception receptors (eg, cooling receptors) and may promote dependence in inexperienced users; (b) acid additives and sugars, which lower the pH of aerosols and shift nicotine from free-base to protonated salt forms, reduce harshness and increase blood nicotine yield; (c) e-cigarette flavourings perceived as sweet or fruity reduce subjective bitterness, increase attractiveness and may escalate use, although their effects on perceived harshness are inconclusive; (d) sugars in tobacco impart sweet sensations, but limited industry-independent data preclude strong conclusions for sugars' roles in inhalation facilitation. Given these findings, WHO policy recommendations suggest that regulators might consider banning ingredients that facilitate inhalation in all commercial inhaled tobacco and nicotine products.

Identification of CDK4/6 Inhibitors as Small Molecule NLRP3 Inflammasome Activators that Facilitate IL-1ß Secretion and T Cell Adjuvanticity.

Several FDA-approved adjuvants signal through the NLRP3 inflammasome and IL-1ß release. Identifying small molecules that induce IL-1ß release could allow targeted delivery and structure-function optimization, thereby improving safety and efficacy of next-generation adjuvants. In this work, we leverage our existing high throughput data set to identify small molecules that induce IL-1ß release. We find that ribociclib induces IL-1ß release when coadministered with a TLR4 agonist in an NLRP3- and caspase-dependent fashion. Ribociclib was formulated with a TLR4 agonist into liposomes, which were used as an adjuvant in an ovalbumin prophylactic vaccine model. The liposomes induced antigen-specific immunity in an IL-1 receptor-dependent fashion. Furthermore, the liposomes were coadministered with a tumor antigen and used in a therapeutic cancer vaccine, where they facilitated rejection of E.G7-OVA tumors. While further chemical optimization of the ribociclib scaffold is needed, this study provides proof-of-concept for its use as an IL-1 producing adjuvant in various immunotherapeutic contexts.

Transoral robotic surgery for pediatric upper airway pathology: An institutional update.

OBJECTIVE: Provide an update on our institution's experience with utilizing transoral robotic surgery (TORS) in pediatric airway surgery and compare these results to surgery by traditional methods. METHODS: Pediatric patients who underwent TORS for treatment of upper airway pathology between 2010 and 2021 at our institution were retrospectively identified and compared to patients with the same or similar pathology who underwent a traditional (open or endoscopic) surgical approach over the same time period. Outcomes of interest included patient demographics, operative times, adverse events, hospital length of stay (LOS), and modified barium swallow (MBSS) results. RESULTS: Forty children (19M, 21F) underwent 46 TORS procedures. Mean age was 6.4 years (range: 6 days-17 years). Most commonly treated pathology included: laryngeal clefts (LC) (n = 18), lymphatic malformations (n = 9), and base of tongue masses (n = 7). Surgical time was decreased in traditional type I LC repairs (mean: 111 vs 149 min, P = 0.04) and lymphatic malformation excisions (59 vs 120 min, p = 0.005). Hospital LOS was increased in TORS type I LC repairs (2.6 vs 1.2 days, P = 0.04). Adverse event rate was similar between TORS and traditional cohorts (17 % vs 16 % cases, P = 0.9). Postoperative MBSS results were improved for TORS type I LC repairs at 6 months (70 % vs 33 %, P = 0.09) and 12 months (82 % vs 43 %, P = 0.05). CONCLUSIONS: Pediatric TORS is practical and safe and has comparable outcomes to traditional surgery. Robotic-assisted LC repair displayed improved postoperative swallow results versus traditional approaches and may be particularly useful in recurrent cases.

Recall of Flavor at First E-cigarette Use and Its Association with E-cigarette Progression: The Mediating Effects of First Sensory Experience.

OBJECTIVE: To test whether the association between flavor at first vape and continued use is mediated through subjective experience at first vape. STUDY DESIGN: In a 2020 cross-sectional survey, 955 young adult ever-vapers recalled their first flavor vaped, subjective experiences at first use, current vaping behavior, nicotine dependence, and quit attempts. A latent class model grouped first-use subjective experiences into classes. Two-part negative binomial hurdle models for each vaping behavior evaluated whether the association of first flavor used with vaping outcomes was mediated by positive experience. RESULTS: Four latent classes (positive, positive and negative, negative, and minimal experience) were further reduced to "any positive experience" (only positive, positive and negative) vs "no positive experience" (negative or minimal). Class membership mediated the association of first flavor used (mint/menthol/ice [ie, "cooling"] or sweet vs other) with each vaping outcome. For example, cooling flavor (vs. other) was associated with positive class membership (OR = 3.5; 95% CI: 1.5, 8.1), which was then associated with any past 30-day vaping (OR = 3.9; 95% CI: 2.7, 5.8) and greater number of vaping days among current vapers (RR = 1.9; 95% CI: 1.3, 2.7) in the two-part hurdle model. Similar results were observed for nicotine dependence and quit attempts, and for sweet (vs other) flavor for any dependence or quit attempts, but not number of dependence symptoms or quit attempts. CONCLUSIONS: Use of a cooling or sweet flavor at first use was associated with having a positive first vaping sensory experience, and then greater likelihood as a young adult of reporting past 30-day vaping, more vaping days, and greater risk for nicotine dependence, suggesting a key mediating role of first use experience.

Para-Hydroxycinnamic Acid Mitigates Senescence and Inflammaging in Human Skin Models.

Para-hydroxycinnamic acid (pHCA) is one of the most abundant naturally occurring hydroxycinnamic acids, a class of chemistries known for their antioxidant properties. In this study, we evaluated the impact of pHCA on different parameters of skin aging in in vitro skin models after H2O2 and UV exposure. These parameters include keratinocyte senescence and differentiation, inflammation, and energy metabolism, as well as the underlying molecular mechanisms. Here we demonstrate that pHCA prevents oxidative stress-induced premature senescence of human primary keratinocytes in both 2D and 3D skin models, while improving clonogenicity in 2D. As aging is linked to inflammation, referred to as inflammaging, we analyzed the release of IL-6, IL-8, and PGE2, known to be associated with senescence. All of them were downregulated by pHCA in both normal and oxidative stress conditions. Mechanistically, DNA damage induced by oxidative stress is prevented by pHCA, while pHCA also exerts a positive effect on the mitochondrial and glycolytic functions under stress. Altogether, these results highlight the protective effects of pHCA against inflammaging, and importantly, help to elucidate its potential mechanisms of action.

Effects of a fruit-ice combination flavor on appeal and sensory experience of vaping and moderation by preexisting e-cigarette flavor preference.

E-cigarettes with ice flavors, which are products with a cooling agent added to a characterizing flavor (e.g., grape-ice), are widely sold. Whether ice flavors appeal to only those who already use them or a to wider population is not widely understood. This secondary analysis of a clinical laboratory experiment tested effects of experimental exposure to e-cigarettes with fruit-ice versus other flavors on the appeal and sensory attributes of vaping and whether fruit-ice effects are moderated by preexisting e-cigarette flavor preference. In a controlled double-blind within-subject randomized online experiment, adults who currently use e-cigarettes, N = 85, M (SD) = 35.5 (13.2) years, 50% female, 54.3% White, self-administered e-liquids varying in flavors (fruit-ice [grape + menthol], tobacco-only, dessert-only [caramel], and fruit-only [strawberry]). Participants rated each product's appeal (liking, disliking, willingness to use again) and sensory attributes (sweetness, smoothness, harshness, bitterness, and coolness). In the overall sample, fruit-ice flavor produced (a) higher appeal, sweetness, smoothness, and coolness and lower bitterness and harshness compared to tobacco and dessert-only flavors and (b) higher coolness and lower sweetness than fruit-only flavors. The appeal-enhancing effect of fruit-ice (vs. tobacco and dessert-only flavors) was stronger among those with preexisting preferences for menthol/mint, fruit, and ice flavors, but not for those who typically used tobacco-only flavors. Our findings suggest that restrictions on fruit-ice-flavored e-cigarettes may reduce the appeal of vaping, particularly among vapers with preferences for products with cooling, fruit, and/or sweet sensory attributes. (PsycInfo Database Record (c) 2024 APA, all rights reserved).

B cell-based therapy produces antibodies that inhibit glioblastoma growth.

Glioblastoma (GBM) is a highly aggressive and malignant brain tumor with limited therapeutic options and a poor prognosis. Despite current treatments, the invasive nature of GBM often leads to recurrence. A promising alternative strategy is to harness the potential of the immune system against tumor cells. Our previous data showed that the Bvax (B-cell-based vaccine) can induce therapeutic responses in preclinical models of GBM. In this study, we aim to characterize the antigenic reactivity of BVax-derived antibodies and evaluate their therapeutic potential. We performed immunoproteomics and functional assays in murine models and human GBM patient samples. Our investigations revealed that BVax distributes throughout the GBM tumor microenvironment (TME) and then differentiates into antibody-producing plasmablasts. Proteomic analyses indicate that the antibodies produced by BVax display unique reactivity, predominantly targeting factors associated with cell motility and the extracellular matrix. Crucially, these antibodies inhibit critical processes such as GBM cell migration and invasion. These findings provide valuable insights into the therapeutic potential of BVax-derived antibodies for GBM patients, pointing towards a novel direction in GBM immunotherapy.

Ghost cells: Wilder Penfield and the characterization of glia and glial pathology, 1924-1932.

Wilder Penfield is known for his contributions to the structure-function relationship of the brain and for the surgical treatment of focal epilepsy. Less well known are his contributions to the study of glial cells and his investigation of their role in human neuropathology. Penfield learned the gold and silver methods for staining neurons, glial cells, and their projections from Charles Sherrington and Pío del Río-Hortega. He and his colleague William Cone established a laboratory for the study of glial cells and human neuropathology using metallic stains, initially at the Presbyterian Hospital in New York City in 1925, and then at the Montreal Neurological Institute in 1928. Penfield, Cone, and their research fellows, building on the findings of Río-Hortega, confirmed the existence of oligodendrocytes and their relationship with myelin, and investigated the putative mesodermal origin of microglia. They discovered the reaction of oligodendrocytes to pathological stressors, and the phagocytic activity of microglia in human gliomas. In this article, we argue that Penfield's studies of astrocytes, oligodendrocytes, and microglia, and their responses to craniocerebral trauma, epilepsy, malignant brain tumors, and other pathologies of the central nervous system inaugurated a new era in clinical neurocytology and neuropathology.

Extent of resection and progression-free survival in vestibular schwannoma: a volumetric analysis.

OBJECTIVE: To preserve facial nerve function in vestibular schwannoma (VS) microsurgery, some have advocated subtotal resection (STR) if the tumor is densely adherent to a thinned facial nerve. The objective of this study was to determine if residual volume is associated with progression and whether there is a threshold residual volume that should be pursued during STR to prevent recurrence. A secondary objective of this study was to determine whether facial nerve function at last follow-up was associated with extent of resection (EOR). METHODS: Clinical and radiographic data were retrospectively collected from the records of 164 patients with VS who underwent resection. Tumor volumes were measured using Visage, and standard statistical methods were used. The House-Brackmann scale was used to assess changes in facial nerve function before surgery and at last follow-up. RESULTS: Sixty-one patients (37%) received gross-total resection (GTR) and 103 (63%) received STR. The median clinical and radiographic follow-ups were 49 and 48 months, respectively. The median residual volume was 0.5 cm3 after STR. Kaplan-Meier actuarial survival analysis revealed a 96.3% 5-year progression-free survival (PFS) rate after GTR, which was greater than that after STR (84.5%, p = 0.03). Recursive partitioning analysis of patients receiving STR revealed a residual volume of 0.60 cm3 as the optimal threshold for recurrence. Patients with residual volume ≥ 0.60 cm3 had a 76.0% 5-year PFS, regardless of adjuvant SRS, which was lower than that for patients undergoing GTR (96.3%) or STR (95.6%) with residual volumes < 0.60 cm3 (p < 0.01). On Cox regression analysis, residual volume ≥ 0.60 cm3 (HR 14.4, p = 0.01) was independently associated with progression, even when accounting for patient age, adjuvant radiosurgery, and preoperative tumor size. In 112 patients with at least 24 months of follow-up after their last treatment, tumor control was achieved in 111 (99.1%) patients at a median last follow-up of 71 months. Worse facial nerve function at the last follow-up was independently associated with prior treatment for VS (adjusted OR 3.7, p = 0.04), but not residual volume cohort or preoperative tumor volume. CONCLUSIONS: Residual volume > 0.60 cm3 after VS resection was independently associated with tumor progression, even accounting for adjuvant SRS. These data support maximizing the EOR during VS surgery, even if GTR cannot be safely achieved.

A Consensus Statement on the Administration of Systemic Bevacizumab in Patients with Recurrent Respiratory Papillomatosis.

OBJECTIVE: To provide detailed guidance on the administration of systemic bevacizumab in patients with recurrent respiratory papillomatosis (RRP) based on a detailed review of the scientific literature and a consensus of experts with real-world clinical experience. METHODS: A bevacizumab consensus working group (N = 10) was composed of adult and pediatric otolaryngologists, adult and pediatric oncologists, and a representative from the RRP Foundation (RRPF), all with experience administering systemic bevacizumab in patients with RRP. After extensive review of the medical literature, a modified Delphi method-based survey series was utilized to establish consensus on the following key areas: clinical and patient characteristics ideal for treatment candidacy, patient perspective in treatment decisions, treatment access, initial dosing, monitoring, guidelines for tapering and discontinuation, and reintensifying therapy. RESULTS: Seventy-nine statements were identified across nine critical domains, and 45 reached consensus [clinical benefits of bevacizumab (3), patient and disease characteristics for treatment consideration (7), contraindications for treatment (3), shared decision-making (incorporating the patient perspective) (5), treatment access (3), initial dosing and administration (8), monitoring (7), tapering and discontinuation (6), and reintensification (3)]. CONCLUSION: This consensus statement provides the necessary guidance for clinicians to initiate systemic administration of bevacizumab and represents a potential paradigm shift toward nonsurgical treatment options for patients with RRP. LEVEL OF EVIDENCE: 5 Laryngoscope, 2024.

Breast Cancer Screening Interval: Effect on Rate of Late-Stage Disease at Diagnosis and Overall Survival.

PURPOSE: Controversy continues regarding the effect of screening mammography on breast cancer outcomes. We evaluated late-stage cancer rate and overall survival (OS) for different screening intervals using a real-world institutional research data mart. METHODS: Patients having both a cancer registry record of new breast cancer diagnosis and prediagnosis screening history between 2004 and 2019 were identified from our institutional research breast data mart. Time interval between the two screening mammograms immediately preceding diagnosis and the time to cancer diagnosis were determined. Screening interval was deemed annual if ≤15 months, biennial if >15 and ≤27 months, intermittent if >27 months, and baseline if only one prediagnosis screen was known. The primary end point was late-stage cancer (TNM stage IIB or worse), and the secondary end point was OS. The association of screening interval and late-stage cancer was analyzed using multivariable logistic regression adjusting for prediagnosis characteristics. Proportional hazards regression was used for survival analysis. Potential lead time was analyzed using survival from a uniform fixed time point. RESULTS: In total, 8,145 patients with breast cancer had prediagnosis screening mammography in the timeframe. The percentage of late-stage cancers diagnosed increased significantly with screening interval with 9%, 14%, and 19% late stages for annual, biennial, and intermittent groups (P < .001), respectively. The trend persisted regardless of age, race, and menopausal status. Biennial and intermittent groups had substantially worse OS than the annual screened group, with relative hazards of 1.42 (95% CI, 1.11 to 1.82) and 2.69 (95% CI, 2.11 to 3.43), respectively, and 1.39 (95% CI, 1.08 to 1.78) and 2.01 (95% CI, 1.58 to 2.55) after adjustment for potential lead time. CONCLUSION: Annual mammographic screening was associated with lower risk of late-stage cancer and better OS across clinical and demographic subgroups. Our study suggests benefit of annual screening for women 40 years and older.

Outpatient surgery for tibial plateau fractures.

PURPOSE: The purpose of this study was to determine the rates of compartment syndrome and other early complications following outpatient open reduction and internal fixation (ORIF) of tibial plateau fractures. METHODS: This was a retrospective cohort at a single US level I academic trauma centre of patients with tibial plateau fractures managed operatively. Inpatients received their definitive ORIF during their index hospital stay and were admitted post-operatively following ORIF. Outpatients were scheduled for ambulatory surgery during definitive ORIF. Exclusion criteria for outpatient surgery included compartment syndrome, polytrauma, open types IIIb/IIIc, and patients who received any internal fixation during index presentation. The primary outcome measure was post-operative compartment syndrome. Secondary outcomes were return to the 90-day return to the ED, 90-day readmission, surgical wound infection, thromboembolism, and 90-day mortality. An intention-to-treat (ITT) and as-treated (AT) analyses were performed. RESULTS: Totally, 71 inpatients and 47 outpatients were included. There were no cases of post-operative compartment syndrome. In the ITT analysis, there were no differences for inpatients vs outpatients for 90-day re-admission (22.5% vs 12.8%, p = 0.275), 90-day return to the ED (35.2% vs 17.0%, p = 0.052), infection (12.7% vs 2.1%, p = 0.094), DVT (7% vs 4.3%, p = 0.819), or PE 1.4% vs 0.0%, p = 1.000). The AT analysis showed a significantly higher 90-day re-admission (26.9% vs 2.5%, p = 0.003) and 90-day ED visit (38.5% vs 7.5%, p = 0.001) rate in the inpatient group. CONCLUSIONS: Appropriately selected patients with isolated tibial plateau fractures can have non-inferior rates of compartment syndrome and post-operative complications when compared to inpatients.

High-throughput genetics enables identification of nutrient utilization and accessory energy metabolism genes in a model methanogen.

Archaea are widespread in the environment and play fundamental roles in diverse ecosystems; however, characterization of their unique biology requires advanced tools. This is particularly challenging when characterizing gene function. Here, we generate randomly barcoded transposon libraries in the model methanogenic archaeon Methanococcus maripaludis and use high-throughput growth methods to conduct fitness assays (RB-TnSeq) across over 100 unique growth conditions. Using our approach, we identified new genes involved in nutrient utilization and response to oxidative stress. We identified novel genes for the usage of diverse nitrogen sources in M. maripaludis including a putative regulator of alanine deamination and molybdate transporters important for nitrogen fixation. Furthermore, leveraging the fitness data, we inferred that M. maripaludis can utilize additional nitrogen sources including ʟ-glutamine, ᴅ-glucuronamide, and adenosine. Under autotrophic growth conditions, we identified a gene encoding a domain of unknown function (DUF166) that is important for fitness and hypothesize that it has an accessory role in carbon dioxide assimilation. Finally, comparing fitness costs of oxygen versus sulfite stress, we identified a previously uncharacterized class of dissimilatory sulfite reductase-like proteins (Dsr-LP; group IIId) that is important during growth in the presence of sulfite. When overexpressed, Dsr-LP conferred sulfite resistance and enabled use of sulfite as the sole sulfur source. The high-throughput approach employed here allowed for generation of a large-scale data set that can be used as a resource to further understand gene function and metabolism in the archaeal domain.IMPORTANCEArchaea are widespread in the environment, yet basic aspects of their biology remain underexplored. To address this, we apply randomly barcoded transposon libraries (RB-TnSeq) to the model archaeon Methanococcus maripaludis. RB-TnSeq coupled with high-throughput growth assays across over 100 unique conditions identified roles for previously uncharacterized genes, including several encoding proteins with domains of unknown function (DUFs). We also expand on our understanding of carbon and nitrogen metabolism and characterize a group IIId dissimilatory sulfite reductase-like protein as a functional sulfite reductase. This data set encompasses a wide range of additional conditions including stress, nitrogen fixation, amino acid supplementation, and autotrophy, thus providing an extensive data set for the archaeal community to mine for characterizing additional genes of unknown function.

Pediatric glioma immune profiling identifies TIM3 as a therapeutic target in BRAF-Fusion pilocytic astrocytoma.

Despite being the leading cause of childhood mortality, pediatric gliomas have been relatively understudied, and the repurposing of immunotherapies has not been successful. Whole transcriptome sequencing, single-cell sequencing, and sequential multiplex immunofluorescence were used to identify an immunotherapy strategy evaluated in multiple preclinical glioma models. MAPK-driven pediatric gliomas have a higher interferon signature relative to other molecular subgroups. Single-cell sequencing identified an activated and cytotoxic microglia population designated MG-Act in BRAF-fused MAPK-activated pilocytic astrocytoma (PA), but not in high-grade gliomas or normal brain. TIM3 is expressed on MG-Act and on the myeloid cells lining the tumor vasculature but not normal brain. TIM3 expression becomes upregulated on immune cells in the PA microenvironment and anti-TIM3 reprograms ex vivo immune cells from human PAs to a pro-inflammatory cytotoxic phenotype. In a genetically engineered murine model of MAPK-driven low-grade gliomas, anti-TIM3 treatment increased median survival over IgG and anti-PD1 treated mice. ScRNA sequencing data during the therapeutic window of anti-TIM3 demonstrates enrichment of the MG-Act population. The therapeutic activity of anti-TIM3 is abrogated in the CX3CR1 microglia knockout background. These data support the use of anti-TIM3 in clinical trials of pediatric low-grade MAPK-driven gliomas.