Assessing Abnormal Proximal Junctional Angles in Adult Spinal Deformity: A Normative Data Approach to Define Proximal Junctional Kyphosis.

STUDY DESIGN: Multicentric retrospective study of prospectively collected data. OBJECTIVE: Based on normative data from a cohort of asymptomatic volunteers, this study sought to determine the rate of abnormal values of proximal junctional angles (PJA) in adult spinal deformity (ASD) surgery patients, and compare it with PJK rate. SUMMARY OF BACKGROUND DATA: Proximal junctional kyphosis (PJK) definition does not take the vertebral level into account. METHODS: This study included 721 healthy volunteers and 824 ASD surgery patients with 2-year postoperative follow-up. Normative values for each disc and vertebral body between T1 and T12 were analyzed, then normative values for PJA at each thoracic level were defined in the volunteer cohort as the mean±2 standard deviations. PJA abnormal values at the upper instrumented vertebra (UIV) were compared with Glattes' and Lovecchio's definitions for PJK in the ASD population at two years. RESULTS: Mean age was 37.7±16.3 in the volunteer cohort, with 50.5% of females. Mean thoracic kyphosis (TK) was -50.9±10.8°. Corridors of normality included PJA greater than 20° between T3 and T12. Mean age was 60.5±14.0 years in the ASD cohort, with 77.2% of females. Mean baseline TK was -37.4±19.9°, with a significant increase after surgery (-15.6±15.3°, P<0.001). There was 46.2% of PJK according to Glattes' versus 8.7% according to Lovecchio's and 22.9% of kyphotic PJA compared to normative values (P<0.001). CONCLUSION: This study provides normative values for segmental and regional alignment of thoracic spine, used to describe abnormal values of PJA for each level. Using level-adjusted PJA values allows a more precise assessment of abnormal proximal angles and question the definition for PJK. LEVEL OF EVIDENCE: II.

Mesenchymal Stromal Cell Implants for Chronic Motor Deficits After Traumatic Brain Injury: Post Hoc Analysis of a Randomized Trial.

BACKGROUND AND OBJECTIVES: Traumatic brain injury (TBI) is frequently characterized by chronic motor deficits. Therefore, this clinical trial assessed whether intracranial implantation of allogeneic modified mesenchymal stromal (SB623) cells can improve chronic motor deficits after TBI. METHODS: Post hoc analysis of the double-blind, randomized, prospective, surgical sham-controlled, phase 2, STEMTRA clinical trial (June 2016 and March 2019) with 48 weeks of follow-up was conducted. In this international, multicenter clinical trial, eligible participants had moderate-to-severe TBI, were ≥12 months postinjury, and had chronic motor deficits. Participants were randomized in a 1:1:1:1 ratio to stereotactic surgical intracranial implantation of SB623 cells (2.5 × 106, 5.0 × 106, 10 × 106) or surgical sham-controlled procedure. The prespecified primary efficacy end point was significantly greater change from baseline of the Fugl-Meyer Motor Scale (FMMS) score, a measure of motor status, for the SB623 pooled vs control arm at 24 weeks. RESULTS: A total of 211 participants were screened, 148 were excluded, and 63 underwent randomization, of which 61 (97%; mean age, 34 [SD, 12] years; 43 men [70.5%]) completed the trial. Single participants in the SB623 2.5 × 106 and 5.0 × 106 cell dose groups discontinued before surgery. Safety and efficacy (modified intent-to-treat) were assessed in participants who underwent surgery (N = 61; SB623 = 46, controls = 15). The primary efficacy end point (FMMS) was achieved (least squares mean [SE] SB623: +8.3 [1.4]; 95% CI 5.5-11.2 vs control: +2.3 [2.5]; 95% CI -2.7 to 7.3; p = 0.04), with faster improvement of the FMMS score in SB623-treated groups than in controls at 24 weeks and sustained improvement at 48 weeks. At 48 weeks, improvement of function and activities of daily living (ADL) was greater, but not significantly different in SB623-treated groups vs controls. The incidence of adverse events was equivalent in SB623-treated groups and controls. There were no deaths or withdrawals due to adverse events. DISCUSSION: Intraparenchymal implantation of SB623 cells was safe and significantly improved motor status at 24 weeks in participants with chronic motor deficits after TBI, with continued improvement of function and ADL at 48 weeks. Cell therapy can modify chronic neurologic deficits after TBI. TRIAL REGISTRATION INFORMATION: ClinicalTrials.gov Identifier: NCT02416492. Submitted to registry: April 15, 2015. First participant enrolled: July 6, 2016. Available at: classic.clinicaltrials.gov/ct2/show/NCT02416492. CLASSIFICATION OF EVIDENCE: This study provides Class I evidence that intracranial implantation of allogeneic stem (SB623) cells in adults with motor deficits from chronic TBI improves motor function at 24 weeks.

Genome-Wide Association Study of Accessory Atrioventricular Pathways.

Importance: Understanding of the genetics of accessory atrioventricular pathways (APs) and affiliated arrhythmias is limited. Objective: To investigate the genetics of APs and affiliated arrhythmias. Design, Setting, and Participants: This was a genome-wide association study (GWAS) of APs, defined by International Classification of Diseases (ICD) codes and/or confirmed by electrophysiology (EP) study. Genome-wide significant AP variants were tested for association with AP-affiliated arrhythmias: paroxysmal supraventricular tachycardia (PSVT), atrial fibrillation (AF), ventricular tachycardia, and cardiac arrest. AP variants were also tested in data on other heart diseases and measures of cardiac physiology. Individuals with APs and control individuals from Iceland (deCODE Genetics), Denmark (Copenhagen Hospital Biobank, Danish Blood Donor Study, and SupraGen/the Danish General Suburban Population Study [GESUS]), the US (Intermountain Healthcare), and the United Kingdom (UK Biobank) were included. Time of phenotype data collection ranged from January 1983 to December 2022. Data were analyzed from August 2022 to January 2024. Exposures: Sequence variants. Main Outcomes and Measures: Genome-wide significant association of sequence variants with APs. Results: The GWAS included 2310 individuals with APs (median [IQR] age, 43 [28-57] years; 1252 [54.2%] male and 1058 [45.8%] female) and 1 206 977 control individuals (median [IQR] year of birth, 1955 [1945-1970]; 632 888 [52.4%] female and 574 089 [47.6%] male). Of the individuals with APs, 909 had been confirmed in EP study. Three common missense variants were associated with APs, in the genes CCDC141 (p.Arg935Trp: adjusted odds ratio [aOR], 1.37; 95% CI, 1.24-1.52, and p.Ala141Val: aOR, 1.55; 95% CI 1.34-1.80) and SCN10A (p.Ala1073Val: OR, 1.22; 95% CI, 1.15-1.30). The 3 variants associated with PSVT and the SCN10A variant associated with AF, supporting an effect on AP-affiliated arrhythmias. All 3 AP risk alleles were associated with higher heart rate and shorter PR interval, and have reported associations with chronotropic response. Conclusions and Relevance: Associations were found between sequence variants and APs that were also associated with risk of PSVT, and thus likely atrioventricular reentrant tachycardia, but had allele-specific associations with AF and conduction disorders. Genetic variation in the modulation of heart rate, chronotropic response, and atrial or atrioventricular node conduction velocity may play a role in the risk of AP-affiliated arrhythmias. Further research into CCDC141 could provide insights for antiarrhythmic therapeutic targeting in the presence of an AP.

FACTORS ASSOCIATED WITH DELAYED DIAGNOSIS IN PATIENTS WITH PRIMARY VITREORETINAL LYMPHOMA.

PURPOSE: To identify demographic and clinical factors associated with delayed diagnosis in patients with primary vitreoretinal lymphoma (VRL). METHODS: Retrospective, tertiary referral center-based cohort study of all patients at Mayo Clinic in Rochester, Minnesota, with a biopsy-proven diagnosis of VRL from January 1, 2000, to October 31, 2022. RESULTS: There were 87 patients included during the 22-year study period with 73 patients (83.9%) diagnosed with VRL upon initial evaluation at the tertiary center, with the other 14 patients (16.1%) diagnosed later. The median referral time was 4.8 months (range: 0-113 months). Patients who received an initial diagnosis of inflammatory uveitis or another incorrect diagnosis elsewhere were referred slower than those initially diagnosed with VRL (P = 0.04). The most common incorrect initial diagnosis from an outside institution was inflammatory uveitis (n = 35, 40.2%). When patients were split into four groups based on referral time, prior use of corticosteroids was associated with a significant delay in referral (P = 0.03). CONCLUSION: Diagnosing VRL continues to be challenging, as months-long delays from initial evaluation to expert referral center evaluation are common. Prior use of corticosteroids was associated with delay in diagnosis and referral time, underscoring the need to increase awareness regarding differences between VRL and uveitis.

Preemptive Approach to Plerixafor Use Is Optimal in Patients With Relapsed/Refractory Germ Cell Tumors Undergoing Peripheral Blood Hematopoietic Stem Cell Collection: Effect on Collection Days, Yields, and Cost.

Evidence describing the use of plerixafor in the off-label population of relapsed/refractory germ cell tumors (GCT) is limited. We aim to describe the effect of rescue versus preemptive plerixafor use on apheresis collection days, collection yields, and cost. We retrospectively collected data on 77 consecutive patients (at least 15 years of age) with GCT who underwent peripheral blood stem cell (PBSC) collection for autologous stem cell transplant between January 1, 2020 and May 1, 2022. Depending on insurance approval, plerixafor was given either as "rescue" (after a first apheresis collection of < 5 × 106 CD34+ cells/kg) or as "preemptive" on Day 4 of granulocyte-colony stimulating factor (G-CSF) prior to the first apheresis collection, if the Day 4 peripheral blood CD34+ count was < 40 cells/µL. A total of 66% of patients who received preemptive plerixafor completed collection in 1 day, similar to good mobilizers who only needed G-CSF (71%, p = 0.366). In contrast, all poor mobilizers in the rescue group required at least 2 days of collection and had lower CD34+ cell yields than the preemptive group (7.15 vs. 9.81 × 106/kg, p = 0.0055). A cost analysis revealed that preemptive plerixafor may save approximately $7000 per patient compared with a rescue approach. Preemptive plerixafor in GCT patients undergoing PBSC collection allows relatively poor mobilizers to collect in fewer days and with lower overall cost. Fewer apheresis procedures result in less risk to the patient, increased patient satisfaction, and the ability to schedule more patients within the constraints of staffing.

Assessing the transportability of radiomic models for lung cancer diagnosis: commercial vs. open-source feature extractors.

Background: Radiomics has shown promise in improving malignancy risk stratification of indeterminate pulmonary nodules (IPNs) with many platforms available, but with no head-to-head comparisons. This study aimed to evaluate transportability of radiomic models across platforms by comparing performances of a commercial radiomic feature extractor (HealthMyne) with an open-source extractor (PyRadiomics) on diagnosis of lung cancer in IPNs. Methods: A commercial radiomic feature extractor was used to segment IPNs from computed tomography (CT) scans, and a previously validated radiomic model based on commercial features was used as baseline (ComRad). Using same segmentation masks, PyRadiomics, an open-source feature extractor was used to build three open-source radiomic models (OpenRad) using different methods: de novo open-source model derived using least absolute shrinkage and selection operator (LASSO) for feature selection, selecting open-source features matched to ComRad features based upon Imaging Biomarker Standardization Initiative (IBSI) nomenclature, and selecting open-source features most highly correlated to ComRad features. Radiomic models were trained on an internal cohort (n=161) and externally validated on 3 cohorts (n=278). We added Mayo clinical risk score to OpenRad and ComRad models, creating integrated clinical radiomic (ClinRad) models. All models were compared using area under the curve (AUC) and evaluated for clinical improvement using bias-corrected clinical net reclassification indices (cNRIs). Results: ComRad AUC was 0.76 [95% confidence interval (CI): 0.71-0.82], and OpenRad AUC was 0.75 (95% CI: 0.69-0.81) for LASSO model, 0.74 (95% CI: 0.68-0.79) for Spearman's correlation, and 0.71 (95% CI: 0.65-0.77) for IBSI. Mayo scores were added to OpenRad LASSO model, which performed best, forming open-source ClinRad model with AUC of 0.80 (95% CI: 0.74-0.86), identical to commercial ClinRad's AUC. Both ClinRad models showed clinical improvement compared to Mayo alone, with commercial ClinRad achieving cNRI of 0.09 (95% CI: 0.02-0.15) for benign and 0.07 (95% CI: 0.00-0.13) for malignant, and open-source ClinRad achieving cNRI of 0.09 (95% CI: 0.02-0.15) for benign and 0.06 (95% CI: 0.00-0.12) for malignant. Conclusions: Transportability of radiomic models across platforms directly does not conserve performance, but radiomic platforms can provide equivalent results when building de novo models allowing for flexibility in feature selection to maximize prediction accuracy.

Regular whole blood donation and gastrointestinal, breast, colorectal and haematological cancer risk among blood donors in Australia.

BACKGROUND AND OBJECTIVES: Several studies have suggested that blood donors have lower risk of gastrointestinal and breast cancers, whereas some have indicated an increased risk of haematological cancers. We examined these associations by appropriately adjusting the 'healthy donor effect' (HDE). MATERIALS AND METHODS: We examined the risk of gastrointestinal/colorectal, breast and haematological cancers in regular high-frequency whole blood (WB) donors using the Sax Institute's 45 and Up Study data linked with blood donation and other health-related data. We calculated 5-year cancer risks, risk differences and risk ratios. To mitigate HDE, we used 5-year qualification period to select the exposure groups, and applied statistical adjustments using inverse probability weighting, along with other advanced doubly robust g-methods. RESULTS: We identified 2867 (42.4%) as regular high-frequency and 3888 (57.6%) as low-frequency donors. The inverse probability weighted 5-year risk difference between high and low-frequency donors for gastrointestinal/colorectal cancer was 0.2% (95% CI, -0.1% to 0.5%) with a risk ratio of 1.25 (0.83-1.68). For breast cancer, the risk difference was -0.2% (-0.9% to 0.4%), with a risk ratio of 0.87 (0.48-1.26). Regarding haematological cancers, the risk difference was 0.0% (-0.3% to 0.5%) with a risk ratio of 0.97 (0.55-1.40). Our doubly robust estimators targeted minimum loss-based estimator (TMLE) and sequentially doubly robust (SDR) estimator, yielded similar results, but none of the findings were statistically significant. CONCLUSION: After applying methods to mitigate the HDE, we did not find any statistically significant differences in the risk of gastrointestinal/colorectal, breast and haematological cancers between regular high-frequency and low-frequency WB donors.

Institutional Conversion to Energy-Efficient Ultra-Low Freezers Decreases Carbon Footprint and Reduces Energy Costs.

Introduction: The storage of biospecimens is a substantial source of greenhouse gas emissions and institutional energy costs. Energy-intensive ultra-low temperature (ULT) freezers used for biospecimen storage are a significant source of carbon emissions. ENERGY STAR-certified ULT freezers have the potential to decrease the carbon footprint. Objective: Quantify the impact of an institutional-scale freezer conversion program on carbon emissions and energy costs. Methods: A ULT freezer energy use prediction model was developed to identify and replace the most inefficient freezers in the research building for this pilot, and eventually institution-wide. Multiple linear regression factors included the number of years of use, storage volume, and ENERGY STAR certification status. Electrical usage and carbon emissions were quantified before and after replacement with ENERGY STAR models. Logistical methods were developed to decrease the risks of exposure of frozen samples to ambient temperature during content transfers. Institution-wide energy costs were derived by converting electrical burden to electrical costs. Carbon footprint assessment from ULT freezer operation was computed using the U.S. EPA Greenhouse Gas Equivalencies Calculator. Results: The pilot project revealed an annual reduction of 310,493 kilowatt hours of electrical usage, equivalent to 134 metric tons of carbon emissions. Annual electrical costs were reduced by $55,889 resulting in an 8-year payback on the initial investment. Using the pilot results, we modeled the benefit of the freezer exchange across the entire institution. The modeling predicted that conversion of the institution's remaining 1119 conventional ULT freezers to ENERGY STAR models would lower annual electrical usage by 7,911,549 kilowatt hours (3423 metric tons of carbon emissions), resulting in savings of over $1.4 million annually. Conclusion: Our methods make a large-scale initiative to replace energy-inefficient ULT freezers logistically possible, reduce carbon footprint, and demonstrate an attractive return on investment while proactively protecting valuable research materials.

Predictive Blood Biomarkers of Targeted Intervention for Chronic Mental Health Symptoms following Traumatic Brain Injury.

The purpose of this study was to assess the performance of predictive blood biomarkers in predicting responsiveness to targeted treatments for chronic psychological issues years after traumatic brain injury (TBI). Targeted Evaluation Action and Monitoring of TBI (TEAM-TBI) was a prospective six-month interventional trial of participants with chronic TBI sequelae (n=95). Plasma biomarkers were analyzed pre-intervention: glial fibrillary acidic protein (GFAP), tau, hyperphosphorylated tau Thr231 (p-Tau), von Willebrand factor (vWF), brain lipid binding protein (BLBP), ubiquitin C-terminal hydrolase-L1 (UCH-L1), vascular endothelial growth factor-a (VEGFa) and claudin-5 (CLDN5). Clinical outcomes included the PTSD Checklist for DSM-5 (PCL-5) and Brief Symptom Inventory-18 (BSI-18). Regression models were built for change in PCL5/BSI-18. Biomarkers and covariates were included. Two models were built to identify responders (improved beyond the minimum clinically important difference). The model to predict change in PCL5 (R2=0.64; p<0.001) included vWF ( p=0.032), BLBP (p=0.001), tau (p=0.002), VEGFa (p=0.015), female sex (p=0.06) and military status (p=0.014).The model to predict change in BSI-18 (R2=0.42; p=0.003) included vWF (p=0.042), VEGFa (p=0.09), BLBP (p=0.01), CLDN5 (p<0.001), female sex (p=0.012), and military status (p=0.004) as predictors. The model to differentiate participants who improved for PCL5 (R2=0.68; p<0.001; AUC=0.93) included vWF (p=0.02), VEGFa (p=0.008), and BLBP (p=0.006). The model to differentiate participants who improved for BSI-18 (R2=0.25; p=0.04; AUC=0.75) included UCH-L1 (p=0.03), GFAP (p=0.06), and vWF (p=0.03). Combinations of pre-intervention blood biomarkers were able to differentiate responders from non-responders in both post-traumatic stress and overall psychological health domains.

Incidence and presenting clinical features of pediatric keratoconus in a US population.

PURPOSE: To report both the incidence of pediatric keratoconus (PKC) in a population-based cohort and the risk for undergoing corneal surgery over a 20-year period at a single institution. METHODS: The medical records of all patients <19 years of age diagnosed with keratoconus while residing in Olmsted County, Minnesota, from January 1, 1975, through December 31, 2019, were retrospectively reviewed. The records of patients <19 years with keratoconus examined at our institution from January 1, 2001, through December 31, 2020, were also reviewed. RESULTS: The incidence of PKC in this population over the 45-year study period was 2.48 cases per 100,000 people per year (95% CI, 1.67-3.29). The mean age at diagnosis was 15.25 years (range, 7-18) years, and 28 (77.8%) were male. During a mean follow-up of 2.8 years (range, 0-17.3 years), 33 of 71 patients managed at our institution (46%) underwent at least one corneal procedure. In this cohort, the Kaplan-Meier risk of requiring a procedure by 10 years following diagnosis was 60%. CONCLUSIONS: The incidence of PKC in Olmsted County, Minnesota, over a 45-year period was 2.48 cases per 100,000 people per year. Nearly half of the patients managed at our institution over the past 20 years required a procedure during follow-up.

Steepness-fragility insights from the temperature derivative analysis of dielectric data of a highly nonsymmetric liquid crystal dimer.

A comprehensive dynamic analysis of the dielectric relaxation-time data across a broad temperature range for both isotropic and nematic phases has been conducted on the CBO3O.Py liquid crystal dimer, the shorter chain-length compound within the highly nonsymmetric pyrene-based series of liquid crystal dimers (CBOnO.Py, with n ranging from 3 to 11). It was known from another previous study that in the nematic phase, three different relaxation processes contribute to the complex dielectric permittivity depending on the orientation of the alignment axis with respect to the probing electric field direction. The temperature-derivative analysis of the relaxation-time data using different analytic functions reveals that the critical-like description, through the dynamic scaling model, best portrays the relaxation-time data in the nematic phase as the system approaches the glass transition. A single glass transition temperature is obtained which is consistent with thermal stimulated depolarization currents experimental determinations published elsewhere. From temperature-dependent steepness index m(T), the activation-critical model is also considered as a more general analytic function from which the dynamic scaling model is a terminal approximation. Additionally, the critical-like parametrization provides insight into obtaining a universal description of the temperature-dependent steepness index m(T), for all liquid crystal compounds belonging to symmetry-selected glass formers, such as rodlike liquid crystal monomers.

Exploring synaptic pathways in traumatic brain injury: a cross-phenotype genomics approach.

Traumatic brain injury (TBI), a global leading cause of mortality and disability, lacks effective treatments to enhance recovery. Synaptic remodeling has been postulated as one mechanism that influences outcomes after TBI. We sought to investigate whether common mechanisms affecting synapse maintenance are shared between TBI and other neuropsychiatric conditions using pathway enrichment tools and genome-wide genotype data, with the goal of highlighting novel treatment targets. We leveraged an integrative approach, combining data from Genome-Wide Association Studies (GWAS) with pathway and gene-set enrichment analyses. Literature review-based and Reactome database-driven approaches were combined to identify synapse-related pathways of interest in TBI outcome, and to assess for shared associations with conditions in which synapse-related pathobiological mechanisms have been implicated, including Alzheimer's disease (AD), schizophrenia (SCZ), major depressive disorder (MDD), post-traumatic stress disorder (PTSD), attention deficit hyperactivity disorder (ADHD) and autism spectrum disorder (ASD). Gene and pathway-level enrichment analyses were conducted using MAGMA and its extensions, e- and H-MAGMA, followed by Mendelian Randomization (MR) to investigate potential causal associations. Of the 98 pathways tested, 32 were significantly enriched in the included conditions. In TBI outcome, we identified significant enrichment in five pathways: "Serotonin clearance from the synaptic cleft" (p-value = 0.0001), "Presynaptic nicotinic acetylcholine receptors" (p-value = 0.0003), "Postsynaptic nicotinic acetylcholine receptors" (p-value = 0.0003), "Highly sodium permeable postsynaptic acetylcholine nicotinic receptors" (p-value = 0.0001), and "Acetylcholine binding and downstream events" pathways (p-value = 0.0003). These associations highlight potential involvement of the cholinergic and serotonergic systems in post-TBI recovery. Three of those pathways were shared between TBI and schizophrenia, suggesting possible pathophysiologic commonalities. In this study we utilize comparative and integrative genomic approaches across brain conditions that share synaptic mechanisms to explore the pathophysiology of TBI outcome. Our results implicate associations between TBI outcome and synaptic pathways as well as pathobiologic overlap with other neuropsychiatric diseases.

Maternal Supplementation with Dietary Betaine during Late Gestation Increased Ewe Plasma Creatine and Lamb Thermoregulation under Field Conditions.

Twin lamb mortality is a significant economic problem impacting the Australian sheep industry. Maternal betaine supplementation improved lamb vigour and early post-natal survival when ewes and lambs were housed indoors, suggesting that betaine may be beneficial to feed under extensive pasture systems. This study investigated whether maternal betaine supplementation during late gestation would improve Merino twin lamb live weight, thermoregulation, vigour and survival to weaning under field conditions. Ewes received dietary betaine at either 0 g/day (CTL; n = 115) or 4 g/day from day 110 of gestation (dG 110) until ~49 days post-partum (pp) (BET; n = 115). Measures indicative of lamb viability and survival were collected within 4-24 h of birth and at ~49 days pp and ~93 days pp. BET ewes had higher creatine and creatinine concentrations at dG 130 than CTL ewes (p < 0.05). BET lambs had a higher rectal temperature within 4-24 h following birth than CTL lambs (p < 0.05). CTL lambs were heavier at ~49 days pp and grew faster from birth to ~49 days pp than BET lambs (both p < 0.05). The time taken after release from the researcher to first suckling was quicker in the CTL lambs than BET lambs (p < 0.05). This study demonstrated that supplementing betaine increased creatine concentration in twin-bearing ewes and thermoregulatory capacity in neonatal lambs under extensive grazing systems.

Radiation immunodynamics in patients with glioblastoma receiving chemoradiation.

Introduction: This is a prospective, rigorous inquiry into the systemic immune effects of standard adjuvant chemoradiotherapy, for WHO grade 4, glioblastoma. The purpose is to identify peripheral immunologic effects never yet reported in key immune populations, including myeloid-derived suppressor cells, which are critical to the immune suppressive environment of glioblastoma. We hypothesize that harmful immune-supportive white blood cells, myeloid derived suppressor cells, expand in response to conventionally fractionated radiotherapy with concurrent temozolomide, essentially promoting systemic immunity similar what is seen in chronic diseases like diabetes and heart disease. Methods: 16 patients were enrolled in a single-institution, observational, immune surveillance study where peripheral blood was collected and interrogated by flow cytometry and RNAseq. Tumor tissue from baseline assessment was analyzed with spatial proteomics to link peripheral blood findings to baseline tissue characteristics. Results: We identified an increase in myeloid-derived suppressor cells during the final week of a six-week treatment of chemoradiotherapy in peripheral blood of patients that were not alive at two years after diagnosis compared to those who were living. This was also associated with a decrease in CD8+ T lymphocytes that produced IFNγ, the potent anti-tumor cytokine. Discussion: These data suggest that, as in chronic inflammatory disease, systemic immunity is impaired following delivery of adjuvant chemoradiotherapy. Finally, baseline investigation of myeloid cells within tumor tissue did not differ between survival groups, indicating immune surveillance of peripheral blood during adjuvant therapy may be a critical missing link to educate our understanding of the immune effects of standard of care therapy for glioblastoma.

Unexpected Expression and Function of FcεRI in Immortalized Breast Cancer Cells: A Cautionary Null Study.

The high-affinity IgE receptor, FcεRI, is typically associated with type 2 effectors such as mast cells (MC). The relatively unique expression profile of FcεRI and accumulating evidence from pre-clinical and clinical settings, such as MC interactions with tumors, have led us to study MCs as a potential therapeutic target in breast cancer. Our work identified MCs interacting with tumor cells at primary sites using the 4T1 (BALB/c) adenocarcinoma model in vivo. However, this analysis was complicated by a surprising finding that the tumor cells intrinsically and strongly expressed FcεRI. We further studied the expression and function of FcεRI in breast cancer cells in vitro. The 4T1 cells expressed FcεRI to a level similar to mouse bone marrow-derived MC (BMMC). Additionally, two established breast cancer cultures derived from human T-47D cells, one estrogen-dependent (E3) and the other estrogen-withdrawn (EWD8), also expressed FcεRI with EWD8 cells showing the greatest abundance. Functional analyses indicated that IgE-mediated antigen stimulation did not elicit classic Ca2+ flux in breast cancer cells as seen in the respective species' MCs; however, FcεRI crosslinking could stimulate IL-6 production from the T-47D derivatives. Preliminary analysis of primary breast cancer biopsy datasets using R2: Genomics Analysis and Visualization Platform was discordant with our in vivo model and in vitro observations. Indeed, FcεRI mRNA abundance declined in metastatic breast cancers compared to non-cancerous breast tissue. Altogether, we report a previously unidentified and immunologically substantive difference between breast cancer models and human primary tumors. Investigators pursuing FcεRI-relevant therapeutics in this context should be aware of this translational barrier.

Single-cell transcriptomics identifies aberrant glomerular angiogenic signalling in the early stages of WT1 kidney disease.

WT1 encodes a podocyte transcription factor whose variants can cause an untreatable glomerular disease in early childhood. Although WT1 regulates many podocyte genes, it is poorly understood which of them are initiators in disease and how they subsequently influence other cell-types in the glomerulus. We hypothesised that this could be resolved using single-cell RNA sequencing (scRNA-seq) and ligand-receptor analysis to profile glomerular cell-cell communication during the early stages of disease in mice harbouring an orthologous human mutation in WT1 (Wt1R394W/+). Podocytes were the most dysregulated cell-type in the early stages of Wt1R394W/+ disease, with disrupted angiogenic signalling between podocytes and the endothelium, including the significant downregulation of transcripts for the vascular factors Vegfa and Nrp1. These signalling changes preceded glomerular endothelial cell loss in advancing disease, a feature also observed in biopsy samples from human WT1 glomerulopathies. Addition of conditioned medium from murine Wt1R394W/+ primary podocytes to wild-type glomerular endothelial cells resulted in impaired endothelial looping and reduced vascular complexity. Despite the loss of key angiogenic molecules in Wt1R394W/+ podocytes, the pro-vascular molecule adrenomedullin was upregulated in Wt1R394W/+ podocytes and plasma and its further administration was able to rescue the impaired looping observed when glomerular endothelium was exposed to Wt1R394W/+ podocyte medium. In comparative analyses, adrenomedullin upregulation was part of a common injury signature across multiple murine and human glomerular disease datasets, whilst other gene changes were unique to WT1 disease. Collectively, our study describes a novel role for altered angiogenic signalling in the initiation of WT1 glomerulopathy. We also identify adrenomedullin as a proangiogenic factor, which despite being upregulated in early injury, offers an insufficient protective response due to the wider milieu of dampened vascular signalling that results in endothelial cell loss in later disease. © 2024 The Author(s). The Journal of Pathology published by John Wiley & Sons Ltd on behalf of The Pathological Society of Great Britain and Ireland.

Population-Based Incidence and Clinical Characteristics of Ocular Adnexal Tumors in Olmsted County, Minnesota.

PURPOSE: To determine the incidence and clinical characteristics of ocular adnexaltumors in Olmsted County, Minnesota. METHODS: Retrospective population-based cohort study of all patients residing in Olmsted County, Minnesota diagnosed with any ocular tumor from January 1, 2006, to December 31, 2015. The medical records of all patients with an incident diagnosis of any ocular adnexal tumor were reviewed using the Rochester Epidemiology Project medical record linkage system for patient demographics, tumor type, and histopathologic confirmation. Incidence rates were calculated per 100,000 person-years. Poisson regression analysis was used to analyze changes in incidence over time. RESULTS: There were 717 patients diagnosed with ocular adnexal tumors during the 10-year study period, yielding an age- and sex-adjusted incidence rate of 59.7 per 100,000 (95% CI 55.4 to 64.0, p < 0.05) per year. In total, 764 tumors were diagnosed. Most tumors were eyelid lesions (N = 756, 99.0%), which were mostly benign (N = 512, 67.8%) with epidermal inclusion cysts (N = 275, 36.0%), hidrocystoma (N = 70, 9.2%), and eyelid sebaceous cysts (N = 46, 6.1%) accounting for the majority. Malignant eyelid lesions (N = 244, 31.9%) were relatively common with basal cell carcinoma (N = 184, 24.1%) and squamous cell carcinoma (N = 49, 6.4%) having the highest frequencies. Orbital tumors (N = 8, 1.0%) were infrequent. Of the orbital tumors, the most common was lacrimal gland adenoidcystic carcinoma (N = 2, 25.0%). CONCLUSIONS: In a population-based setting, most ocular adnexal tumors were benign eyelid lesions. Understanding the epidemiology of ocular adnexal tumors is important to aid providers in diagnosing and facilitating appropriate referrals of potentially vision- and life-threatening malignancies.