Vehicle Control Research Articles

Variational resolution probability maps for multi-UAV search tasks with changeable searching behaviors

Cooperative search is a fundamental task in the autonomous control of multiple unmanned aerial vehicles (UAV) and much effort has been spent on finding an effective way to improve the global search performances. Two behavioral aspects are essentially considered during the multi-UAV cooperative search. One is the coverage (exploration) and the other is the retrospect to hotspot regions (exploitation). Most existing studies focus on the solutions to the former issue, by using the cognitive maps with fixed grid layouts, while ignoring the coexistence of both sides, which potentially induces the failures of finding hidden and indetectable targets. This paper presents a novel search method based on the cognitive maps with alternative grid settings, namely the variational resolution probability map (VRPM), to support the global searches with changeable phases and behaviors. To realize the transformations between different resolution maps in a given budget, the incremental learning is used to update the VRPM under limited number of samples. Potential games with different objectives are used to model the multi-UAV searching behaviors during explorations and exploitations. Lyapunov vector field (LVF) is then used to steer the UAVs toward their target grids in a given time horizon, so that the control period can be properly coincided with the temporal sequences of decision makings. Simulation results demonstrate the phase transitions from the explorations to the exploitations by the proposed method. Search accuracy and the retrospects to hotspot areas are also improved by comparison with the existing search strategies.

SEPIC Based BLDC Motor Control for Electric Vehicles

Abstract: This paper presents a BLDC motor that can be utilized in electric vehicle applications where speed control is necessary. It is driven by a single-ended primary inductive converter (SEPIC). The inverter that feeds the motor uses PWM-based switching; however, a SEPIC converter in the DC link can be used to adjust the voltage level. Electronic commutation gating pulses are provided to the inverter input and the DC input. It is effective for electric vehicles since it uses a converter that switches to manage voltage. The SEPIC is lightweight and suitable for use with small electric cars. This paper describes using a SEPIC converter to drive a BLDC motor. The BLDC motor's closed-loop speed control is modelled using MATLAB /SIMULINK, and the simulation's outcomes are presented.

Abstract C016: Age dictates immune response within the tumor microenvironment of triple negative breast cancer

Abstract Age is the biggest risk factor for developing most forms of cancer. In breast cancer, over 50% of patients are over 60 years old; however, less than 25% of patients enrolled in clinical trials are in this age bracket. Furthermore, most mouse modeling is performed in young mice, meaning that from preclinical through clinical stages patients ages are not accurately reflected in trials designs. Triple negative breast cancer (TNBC) is an aggressive subtype of breast cancer with no targeted therapies, so many treatment strategies utilize chemotherapy in combination with immunotherapy – which relies on a functional immune response. It is well known that immune function declines as we age, and thus therapeutic responses may differ with age. It is yet unclear how the tumor immune microenvironment changes with age, and how this may impact therapeutic response. Here, we utilize a mouse model which mimics hallmarks of human immune aging to understand how age impacts TNBC immune responses to dictate disease progression and therapeutic response. Immunologically young (8 weeks old) and aged (∼12 months old) FVB mice were orthotopically injected with syngeneic Met1 TNBC cells. Once tumors became palpable, mice were treated with paclitaxel chemotherapy or vehicle control and/or anti-PDL1 immunotherapy to mimic first-line therapeutic strategies. Tumors grew significantly faster in young control mice compared to aged, but young mice had a more robust response to therapy. Notable, only young mice had a significant benefit of combo therapy over monotherapy alone. Despite aged mice having more circulating CD8+ and CD4+ T cells, tumors in aged mice had significantly more infiltration of CD8+ cytotoxic T cells and FOXP3+ inhibitory T-regulatory cells compared to young control mice, yet only tumors in young mice exhibited an increase in T cell infiltration with therapy. Additionally, the ratio of CD8+ T cells and FOXP3+ Tregs was unchanged with age or treatment. Depletion of either CD8+ or CD4+ T cells led to a complete loss of age-associated differences in tumor growth. RNA sequencing analysis suggests that infiltrating T cells in aged mice may be exhausted due to a chronic interferon (IFN) phenotype, indicated by enrichment of IFN-alpha, IFN-gamma, and IL2 pathways, as well as increased expression of immune exhaustion markers PD1, CTLA4, TIM3 and LAG3. Neutralization of IFN-gamma blocks therapeutic response to anti-PDL1 in young mice but has no impact on aged mice. Analysis of published single cell RNAseq data indicated the same IFN response pathways enriched with age in human TNBC. These results demonstrate that the aging tumor immune microenvironment significantly impacts tumor progression and therapeutic response and suggest that targeting IFN signaling may lead to improved therapeutic responses. Together these findings establish a significant impact of immune aging on TNBC disease progression and response to therapy, highlighting the importance of developing age- stratified treatment strategies to improve responses in patients of all ages. Citation Format: Milos Spasic, Adrienne Parsons, Abigail Recko, Elizabeth Mittendorf, Peter van Galen, Sandra McAllister. Age dictates immune response within the tumor microenvironment of triple negative breast cancer [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Tumor-body Interactions: The Roles of Micro- and Macroenvironment in Cancer; 2024 Nov 17-20; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(22_Suppl):Abstract nr C016.

Abstract B048: Neutrophil extracellular traps-targeting DNase I enhances the therapeutic efficacy of CAR-T cell adoptive immunotherapy in a syngeneic murine metastasis model

Abstract In recent years, chimeric antigen receptor (CAR) T cell therapy has emerged as a promising approach for treating various malignancies, most notably in hematologic cancers. However, its efficacy in solid tumors is often hindered by an immunosuppressive tumor microenvironment (TME), which, in particular, is frequently characterized by the presence of tumor-associated cell-free DNA (cfDNA), especially in the form of neutrophil extracellular traps (NETs). In adoptive immunotherapies NETs, composed primarily of neutrophil-derived DNA, histones and neutrophil granule enzymes (e.g., MPO, NE), act as physical barriers and induce the secretion of immunosuppressive factors that impair CAR T cell function. This study investigates the potential of co-administration of deoxyribonuclease I (DNase I) with CAR T cells in a syngeneic B16 melanoma murine model of lung metastasis. Bioluminescent imaging of melanoma metastatic processes has shown that a single injection of DNase I (10 mg/kg) suppresses B16-EGFR lung metastasis at early stages in comparison to the vehicle control group but does not improve survival. However, co-administration of DNase I with murine EGFR-CAR T cells significantly suppresses tumor burden, decreases the number of metastatic foci, and substantially prolongs survival compared to the CAR T cell monotherapy group. Degrading of NETs [RB1] by DNase I increases the amount of tumor-infiltrating T and CAR T cells and reduces the immunosuppressive effects of the TME. This is reflected in the CD8 population of tumor-infiltrating CAR T cells from the DNase I treated group, which have lower expression of PD-1 and TIM-3 exhaustion markers. This research highlights the critical role of the NETs in modulating CAR T cell efficacy and provides a compelling rationale for incorporating DNase I as an adjunctive treatment to improve therapeutic responses in patients undergoing CAR T cell therapy. However, further clinical studies are warranted to validate these findings and explore the translational potential of this combinatorial approach in enhancing cancer treatment. [RB1]Again - this takes the emphasis and the reader away from NETs and into the cfDNA universe. Citation Format: Alexey Stepanov, Wenjian Wang, Yingqin Hou, Ivan Chernikov Ivan Chernikov, Reid Bissonnette, George Tetz, Grigory Borisenko, Dmitry Genkin. Neutrophil extracellular traps-targeting DNase I enhances the therapeutic efficacy of CAR-T cell adoptive immunotherapy in a syngeneic murine metastasis model [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Tumor-body Interactions: The Roles of Micro- and Macroenvironment in Cancer; 2024 Nov 17-20; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(22_Suppl):Abstract nr B048.

Model-informed drug development for antimicrobials: translational pharmacokinetic-pharmacodynamic modelling of apramycin to facilitate prediction of efficacious dose in complicated urinary tract infections.

The use of mouse models of complicated urinary tract infection (cUTI) has usually been limited to a single timepoint assessment of bacterial burden. Based on longitudinal in vitro and in vivo data, we developed a pharmacokinetic-pharmacodynamic (PKPD) model to assess the efficacy of apramycin, a broad-spectrum aminoglycoside antibiotic, in mouse models of cUTI. Two Escherichia coli strains were studied (EN591 and ATCC 700336). Apramycin exposure-effect relationships were established with in vitro time-kill data at pH 6 and pH 7.4 and in mice with cUTI. Immunocompetent mice were treated with apramycin (1.5-30 mg/kg) starting 24 h post-infection. Kidney and bladder tissue were collected 6-96 h post-infection for cfu determination. A PKPD model integrating all data was developed and simulations were performed to predict bacterial burden in humans. Treatment with apramycin reduced the bacterial load in kidneys and bladder tissue up to 4.3-log compared with vehicle control. In vitro and in vivo tissue time-course efficacy data were integrated into the PKPD model, showing 76%-98% reduction of bacterial net growth and 3- to 145-fold increase in apramycin potency in vivo compared with in vitro. Simulations suggested that an 11 mg/kg daily dose would be sufficient to achieve bacterial stasis in kidneys and bladder in humans. PKPD modelling with in vitro and in vivo PK and PD data enabled simultaneous evaluation of the different components that influence drug effect, an approach that had not yet been evaluated for antibiotics in the cUTI model and that has potential to enhance model-informed drug development of antibiotics.

Ruxolitinib Cream Monotherapy Improved Symptoms and Quality of Life in Adults and Adolescents with Mild-to-Moderate Atopic Dermatitis: Patient-Reported Outcomes from Two Phase III Studies.

Atopic dermatitis (AD) is associated with itch, skin pain, sleep disturbances, and diminished quality of life (QoL). Ruxolitinib (Janus kinase [JAK] 1/JAK2 inhibitor) cream demonstrated efficacy and safety in adults and adolescents with mild-to-moderate AD in two phase III studies (TRuE-AD1/TRuE-AD2). In TRuE-AD1/TRuE-AD2, significant improvements in itch were observed as early as 12 h following application of ruxolitinib cream. The aim of this paper was to assess additional patient-reported outcomes (PROs) in the vehicle-controlled (VC) and long-term safety (LTS) periods of TRuE-AD1/TRuE-AD2. In the TRuE-AD studies, patients aged ≥12 years with AD were randomized 2:2:1 to apply twice-daily 1.5% ruxolitinib cream, 0.75% ruxolitinib cream, or vehicle cream continuously for 8weeks (VC period). During the LTS period, patients applied the same ruxolitinib cream strength, but on an as-needed basis; patients who initially applied vehicle were re-randomized to apply 0.75% or 1.5% ruxolitinib cream. Pooled data from both study periods were analyzed. PRO assessments included symptoms (itch [Patient-Oriented Eczema Measure, POEM], skin pain [numerical rating scale], and sleep [POEM and Patient-Reported Outcomes Measurement Information System]) and assessments of disease-specific QoL (Dermatology Life Quality Index [DLQI] and the children's version [CDLQI]). A total of 1208 and 1031 patients from the VC and LTS periods, respectively, were included in the analysis. Significant improvements in skin pain were observed within 12h among patients who applied ruxolitinib cream versus vehicle; improvements continued throughout the VC period. Improvements in patient-reported symptoms (including sleep) were observed within 2 weeks (first assessment) of ruxolitinib cream application. At Week 2, significant improvements in symptom burden and overall QoL were observed with ruxolitinib cream (0.75%/1.5%) versus vehicle in POEM (-8.9/-9.8 vs -2.2; both p<0.0001), DLQI (mean changes from baseline, -5.8/-6.1 vs -1.2; both p<0.0001), and CDLQI (-4.3/-5.3 vs -1.3; both p<0.0001). Further symptom burden and QoL improvements were reported during the VC period and were maintained through the end of the LTS period (Week 52). Consistent with the previously reported itch response data, ruxolitinib cream improved skin pain within 12 h of application. Ruxolitinib cream improved patient-reported AD symptom burden and overall QoL by Week 2. Improvements continued or were maintained for 52 weeks. (Graphical abstract and plain language summary available). ClinicalTrials.gov identifiers, NCT03745638 and NCT03745651 (both studies were registered on November 19, 2018).

Modelling, Simulation and Testing of Steer-by-Wire System with Variable Steering Ratio Control Strategy in 14-DOF Full Vehicle Model

This research explores the application of Proportional-Integral-Derivative (PID) controllers and Variable Steering Ratio (VSR) control strategies within a Steer-by-Wire (SbW) system, utilizing a comprehensive 14 Degrees of Freedom (DOF) full vehicle dynamic model. The study aims to advance vehicle dynamics and control through SbW technology. This research addresses the challenge of optimizing steering control by integrating an SbW system with PID and VSR strategies to overcome the limitations of traditional steering mechanisms and enhance vehicle responsiveness and stability. The SbW system, using a PID-based controller, was rigorously tested and validated using MATLAB Simulink and Hardware-in-the-Loop System (HiLS) method. The performance of the system was assessed through four distinct signal tests: step, sine, square, and sawtooth inputs. These tests confirmed the precision of the PID controller in position tracking. To improve the practical applicability of the SbW system, a 14-DOF vehicle dynamic model was developed and validated before integration with the SbW system. The integrated system accurately replicated conventional steering behavior, as validated by Step Steer Input (SSI) and Double Lane Change (DLC) tests, demonstrating less than 11% RMS percentage error - well within the acceptable threshold of less than 15%. Additionally, the study proposed and validated VSR control strategies through the DLC test. Results indicate that the PID controller effectively tracked desired signals, while the VSR strategy reduced driver workload and improved steering stability. This research offers valuable insights into enhancing SbW system performance and lays a foundation for future advancements in vehicle control systems.

Model Reference Adaptive Control of an Independent Steer-by-Wire System: A Simulation Using a 14-Degree-of-Freedom Vehicle Model

This research delves into how Model Reference Adaptive Control (MRAC) can be applied in an independent Steer-by-Wire (SBW) system, utilising a detailed 14 Degrees of Freedom (DOF) full-vehicle model. This study is all about pushing forward vehicle dynamics and control using SBW technology. This study also has come up with some cutting-edge control algorithms that allow each wheel to be steered independently, which seriously boosts how manoeuvrable and responsive the vehicle is. Through simulations, the study shows that MRAC is a big improvement over traditional control methods where quantitative analysis shows that MRAC reduces yaw rate errors by up to 66.67% compared to Proportional-Integral-Derivative (PID) and 50% compared to Multi-order PID (MOPID). Additionally, in lateral acceleration and sideslip angle controls, MRAC demonstrates a similar reduction in errors, significantly outperforming PID and MOPID with errors maintained well below 10%, proving its worth in predictive control and real-time adaptability to various road conditions and driver intentions. The key finding from this study is that MRAC greatly enhances manoeuvrability and responsiveness compared to standard methods which offer flexibility according to the different driving scenarios. There are notable advancements in vehicle steering systems, which contribute to safer and more efficient driving. In essence, this work marks a significant step forward in automotive steering technology, opening the door to safer and more efficient modern vehicles.

Efficacy of chitosan root canal medicament against cross-kingdom dual-species biofilm of Candida albicans and Enterococcus faecalis in an in vitro root-canal model.

Infection of the root canals with cross-kingdom biofilm of Candida albicans (C. albicans) and Enterococcus faecalis (E. faecalis) causes more inflammation and is resistant to conventional medication. This study aimed to evaluate the efficacy of chitosan paste (chitosan + propylene glycol (PG)) against this dual-species biofilm in comparison to calcium hydroxide (Ca(OH)2) in the root canals of human teeth. Fifty extracted single-rooted human teeth were prepared and inoculated with C. albicans and E. faecalis, and incubated for 3days for biofilm formation. Four experimental groups (N = 10 each) were applied with different root canal medications for 7days, including (1) No medication (negative control); (2) Ca(OH)2 + distilled water; (3) PG (vehicle control) and (4) chitosan + PG. The efficacy of root canal medicaments was assessed by determining the percentage of remaining colony-forming units (CFUs) of viable microorganisms in intracanal dentin. The dentin was collected, serially diluted, and spread on solid media for colony count. The biofilm structure in root canals at 3days and 10days (ie. before and after treatments) were observed with scanning electron microscopy (SEM) at 5000× and 10,000× magnifications. Data were analyzed using Welch's ANOVA and Games-Howell post-hoc test (α = 5%). The results show that Chitosan + PG group had a significantly lower percentage of remaining microorganisms when compared to PG, Ca(OH)2, and negative control groups (P-value = 0.015, 0.005, and < 0.001, respectively). Consistently, SEM showed the lowest amount of remaining biofilm in Chitosan + PG group. Thus, Chitosan + PG paste was the most efficacious root canal medicament against dual-species biofilm of C. albicans and E. faecalis in this model.

Assessing cytotoxicity: a comparative analysis of biodegradable and conventional 3D-printing materials post-steam sterilization for surgical guides

Introduction.Ecological concerns and the depletion of petroleum resources have driven the exploration of biodegradable 3D-printing materials derived from bio-renewable sources, such as polylactic acid (PLA) and polyhydroxyalkanoates (PHA). This study aimed to compare the potential cytotoxic effects of a biodegradable PLA/PHA blend filament, a conventional photopolymer (MED610), and a combination of MED610 with a support material (SUP705) before and after steam sterilization in vitro, with a focus on their application in the production of surgical guides.Materials and Methods.PLA/PHA, MED610, and SUP705 (both in their pure and steam-sterilized forms;n= 6 per group) were assessed for their cytotoxic effects on human fibroblasts using the neutral red uptake assay. Positive controls included zinc diethyldithiocarbamate and zinc dibutyldithiocarbamate, while high-density polyethylene served as a negative control. A stock solution of the extraction medium was used as the vehicle control (VC).Results.Significant differences in cell viability were observed between pure PLA/PHA (1.2 ± 0.24) and MED610 (0.94 ± 0.08) (p= 0.005). However, both materials exhibited non-cytotoxicity, with cell viability exceeding 70% compared to VCs. SUP705 (0.58 ± 0.42) demonstrated significantly reduced cell viability compared to PLA/PHA (p= 0.001) and MED610 (p= 0.007). After steam sterilization, no significant difference in cell viability was noted between MED610 (1.0 ± 0.08) and PLA/PHA (1.2 ± 0.25) (p= 0.111). While both materials remained non-cytotoxic after sterilization, SUP705 (0.60 ± 0.45) exhibited cytotoxic effects compared to MED610 (p= 0.006) and PLA/PHA (p< 0.001). Steam sterilization did not induce significant cytotoxic effects in the investigated materials (p= 0.123).Conclusion.Pure and steam-sterilized PLA/PHA and MED610 were not cytotoxic, supporting their potential use in the production of surgical guides. However, the observed cytotoxicity of SUP705 suggests caution in scenarios requiring sterile conditions, as the removal of support material from complex printed parts may be challenging. The consideration of PLA/PHA is recommended in such settings to ensure biocompatibility.

Novel Inhibitory Actions of Neuroactive Steroid [3α,5α]-3-Hydroxypregnan-20-One on Toll-like Receptor 4-Dependent Neuroimmune Signaling

The endogenous neurosteroid (3α,5α)-3-hydroxypregnan-20-one (3α,5α-THP) modulates inflammatory and neuroinflammatory signaling through toll-like receptors (TLRs) in human and mouse macrophages, human blood cells and alcohol-preferring (P) rat brains. Although it is recognized that 3α,5α-THP inhibits TLR4 activation by blocking interactions with MD2 and MyD88, the comprehensive molecular mechanisms remain to be elucidated. This study explores additional TLR4 activation sites, including TIRAP binding to MyD88, which is pivotal for MyD88 myddosome formation, as well as LPS interactions with the TLR4:MD2 complex. Both male and female P rats (n = 8/group) received intraperitoneal administration of 3α,5α-THP (15 mg/kg; 30 min) or a vehicle control, and their hippocampi were analyzed using immunoprecipitation and immunoblotting techniques. 3α,5α-THP significantly reduces the levels of inflammatory mediators IL-1β and HMGB1, confirming its anti-inflammatory actions. We found that MyD88 binds to TLR4, IRAK4, IRAK1, and TIRAP. Notably, 3α,5α-THP significantly reduces MyD88-TIRAP binding (Males: −31 ± 9%, t-test, p &lt; 0.005; Females: −53 ± 15%, t-test, p &lt; 0.005), without altering MyD88 interactions with IRAK4 or IRAK1, or the baseline expression of these proteins. Additionally, molecular docking and molecular dynamic analysis revealed 3α,5α-THP binding sites on the TLR4:MD2 complex, targeting a hydrophobic pocket of MD2 usually occupied by Lipid A of LPS. Surface plasmon resonance (SPR) assays validated that 3α,5α-THP disrupts MD2 binding of Lipid A (Kd = 4.36 ± 5.7 μM) with an inhibition constant (Ki) of 4.5 ± 1.65 nM. These findings indicate that 3α,5α-THP inhibition of inflammatory mediator production involves blocking critical protein-lipid and protein-protein interactions at key sites of TLR4 activation, shedding light on its mechanisms of action and underscoring its therapeutic potential against TLR4-driven inflammation.

Four-Week GLP Immunotoxicity Assessment of Lactoferrin Alpha Produced by Komagataella phaffii in Sprague-Dawley Rats.

Oral toxicity and toxicokinetic properties of human lactoferrin (LF) alpha produced in Komagataella phaffii (effera™) were investigated in adult Sprague-Dawley rats over a 28-day period under good laboratory practice conditions. Main study dosing used groups of 10 rats/sex/dose, and a secondary study evaluating toxicokinetic parameters used 6 rats/sex/dose. The vehicle control group received sodium citrate buffer, test groups received daily doses of 200, 600, and 2000mg of effera™ per kg body weight, and the comparative control group received 2000mg bovine LF (bLF)/kg body weight per day. T-cell-dependent antibody response against keyhole limpet hemocyanin and immunophenotyping of the spleen were performed as measures of immunotoxicity. Clinical observations, body weight, hematology, coagulation, clinical chemistry, urinalysis, immunotoxicity, gross necropsy, and histopathology were assessed. Toxicokinetic parameters were analyzed as an indication of LF bioavailability, and anti-LF antibody assays were conducted to detect antibodies produced against LF to measure immunogenicity. No treatment related toxicologically significant changes were observed. Based on the absence of toxicologically relevant changes, effera™ is well tolerated in rats at doses up to 2000mg rhLF/kg/day, an amount ∼400 times that of the estimated daily intake at the 90th percentile proposed for human adult use.

Abstract 4141446: Sodium Glucose Co-transporter 2 (SGLT2) Inhibitors Promote Resiliency to High Pressure Stress in the Human Microvasculature

Emerging evidence suggests that vascular stress from cardiovascular-related co-morbidities promotes microvascular dysfunction, a key component in the development of heart failure with preserved ejection fraction. The sodium glucose co-transporter 2 (SGLT2) inhibitor empagliflozin has been shown to reduce both morbidity and mortality associated with heart failure with preserved ejection fraction, however the full scope of influence of this therapy on human microvascular function remains unknown. We hypothesized that pre-treatment of isolated human microvessels with empagliflozin will prevent stress-induced endothelial dysfunction as evidenced by preserving both the magnitude of flow-induced dilation (FID) as well as the ability to dilate to nitric oxide. Human resistance arterioles (80-250µm) from healthy adults (defined as patients with ≤1 risk factor for cardiovascular disease) were dissected from discarded surgical adipose tissue and treated with empagliflozin (1µM), or vehicle control (ethanol) for 16-20 hours prior to the flow experiment. Vessels were cannulated for videomicroscopy and subjected to high intraluminal pressure (150mmHg, 30 min), an acute stress known to induce endothelial dysfunction. Vessels were pre-constricted with endothelin-1 prior to initiation of flow. A nonlinear logistic regression was used to determine differences between curves. Compared to vehicle control, vessels pre-treated with empagliflozin (1µM ) exhibited nitric oxide-dependent FID as dilation was impaired in the presence of the nitric oxide synthase inhibitor L-NAME (EC50 Control: 10.7 vs L-NAME 83.45, p=0.0107). This data suggests that empagliflozin, an SGLT2 inhibitor, promotes microvascular resilience to stress via preservation of nitric oxide-mediated FID. The ability to elicit stress resilience may explain in part some of the cardiovascular benefits associated with SGLT2 inhibitors and may offer unique opportunities for early intervention or prevention of microvascular dysfunction associated with comorbidities that contribute to heart failure with preserved ejection fraction.

Abstract 4146302: NETosis Attenuating Nanoprobe Improves Survival after Cardiac Arrest

NETosis, the process of neutrophil cell death due to the formation and release of neutrophil extracellular traps (NETs) has been implicated in the pathogenesis of heart disease. NETs are composed of immunogenic DNA fragments and citrullinated histones. The role of NETosis in the context of cardiac arrest and resuscitation (CA/R) is however unknown. We previously reported the development of a Dextran-Thiazole Orange (DTO) nanoprobe, with nanomolar affinity for nucleic acids, that exerts acute anti-inflammatory effects and reduces ischemia-reperfusion injury in mice. We aimed here to determine whether the DTO nanoprobe would also bind to the nucleic acid in NETS, whether DTO could attenuate the intensity of NETosis, and whether this would improve outcomes and survival after cardiac arrest. C57Bl6 mice were arrested for 8 minutes before resuscitation. DTO or unmodified dextran (vehicle control) was injected at the time of resuscitation, and again 4 hours later (n=38 mice). Survival and neurological function were scored daily. Time-domain resolved (lifetime) and conventional fluorescence imaging was performed at 4 hours after resuscitation in 6 additional mice injected with DTO. NETosis was evaluated by protein blot in the hearts of cardiac arrest mice injected with DTO or control. DTO significantly (p&lt;0.05) improved 10-day survival (Fig. A), mean survival (from 4 to 7 days, Fig. B), and neurological function (Fig. C) following CA/R. Lifetime imaging (Fig. D), but not conventional fluorescence imaging (Fig. E) revealed the presence of DTO in the injured heart, liver and kidneys. In the heart, DTO signal was significantly increased in CA/R compared to sham mice injected with DTO or CA/R mice injected with control dextran (Fig. F). Citrullinated H3 and HMGB-1 (High mobility group box 1, a chromatin protein) were both upregulated in CA/R, and were significantly attenuated by DTO injection (Fig. G-I). We show for the first time that NETosis plays a key role in cardiac arrest and resuscitation. DTO is capable of detecting NETosis as well as attenuating it, likely via a reduction in HMGB-1 activity. The modulation of NETosis may provide a new avenue to improve survival after cardiac arrest.

Adaptive Fuzzy Optimal Control of Multiple Autonomous Surface Vehicles with Uncertain Dynamics

Adaptive Fuzzy Optimal Control of Multiple Autonomous Surface Vehicles with Uncertain Dynamics

Abstract 4138674: Long-acting CRF2 receptor agonist, COR-1389, improves cardiopulmonary function in the rat model of Sugen plus hypoxia-induced pulmonary hypertension and right heart failure

Introduction: Urocortin-2 (UCN-2), a peptide which is part of the corticotropin-releasing factor (CRF) family, functions as an autocrine and paracrine factor, exerting its effects on cardiac and pulmonary function through agonism of CRF2 receptors. Although acute administration of UCN-2 has shown promise by improving heart and lung function in conditions like heart failure (HF) and pulmonary hypertension (PH), its limited stability impedes its chronic therapeutic use. Hypothesis: In this study, we explored the efficacy of COR-1389, a potent, selective and long-acting CRF2 agonist peptide, in the Sugen 5416 (VEGFR2 inhibitor, Su) combined with hypoxia (Hx) rat model of PH and right heart failure (RHF). Methods: To this aim, male adult Sprague Dawley rats were divided into three groups: Control rats (normoxia) were compared with rats injected subcutaneously with 20 mg/kg Sugen 5416 and exposed to chronic hypoxia for 3 weeks, followed by 2 weeks of normoxia. At 5 weeks, control rats and one SuHx group received subcutaneously vehicle (control and SuHx), while the third group received COR-1389 at a dose of 100 μg/kg every 4 days subcutaneously for 3 weeks (SuHx + COR-1389). At 8 weeks, cardiac and pulmonary hemodynamic functions of the three groups were evaluated using echocardiography and right heart catheterization, and heart and lung tissue were evaluated by histology and immunohistochemistry. Results: Compared to controls (n=10), SuHx exhibited increased mean pulmonary arterial pressure (mPAP), right ventricle (RV) hypertrophy (Fulton Index/body weight), muscularization of distal pulmonary arteries, RV fibrosis and cardiomyocyte hypertrophy, alongside reduced RV systolic function (Tricuspid Annular Plane Systolic Excursion, TAPSE) and cardiac output (CO). Conversely, compared to the SuHx + vehicle group (n=11), curative treatment with COR-1389 for 3 weeks in SuHx rats (n=13) led to enhanced RV systolic function (TAPSE) and CO, together with reductions in mPAP, RV hypertrophy, muscularization of pulmonary arteries, RV fibrosis and cardiomyocyte hypertrophy. Systolic blood pressure remained unchanged across all groups. Conclusion: These findings indicate that COR-1389 ameliorates the deteriorating cardiopulmonary parameters observed in the SuHx model and represents a promising approach for the treatment of PH and RHF.

Abstract 4125938: Novel Surgical Model of Thoracic Aortic Aneurysm Mediated by Vasa Vasorum Inhibition

Objective: The adventitial layer of the aorta contains the network of microvessels known as vasa vasorum. Previous work by our team revealed deficient vasa vasorum associated with medial hypoxia and upregulated adventitial thrombospondin-1 (TSP-1), an anti-angiogenic molecule, in human aortic aneurysm. We hypothesize that heightened TSP-1 in the ascending thoracic aorta induces aneurysmal degeneration through its anti-angiogenic effects on the vasa vasorum. We tested this hypothesis in a rabbit model via peri-adventitial delivery of ABT-510, a thrombospondin peptide mimetic. Methods: The ascending aorta was exposed in New Zealand White rabbits (n=16) and treated with ABT-510 proximally and a vehicle control distally. Echocardiography was undertaken immediately prior to the procedure and 14 days post-operatively. Adventitial vasa vasorum were quantified from H&amp;E-stained sections. Elastin and collagen organization were assessed with Verhoeff-Van Gieson (VVG) and pentachrome staining as well as multiphoton microscopy. Immunodetection of Hypoxyprobe™ and alpha smooth muscle actin (αSMA) were also undertaken to identify hypoxia and smooth muscle cell content, respectively. Results: Echocardiography revealed an increase in aortic diameter over 14 days with ABT-510 treatment compared with control (42.27% ± 5.54 vs 20.99% ± 4.10, p=0.004). Histological analysis showed decreased vasa vasorum count (44.7 ± 9.19 vs 85.8 ± 12.89, p=0.048) and decreased vasa vasorum indexed cumulative area (0.013 ± 0.0037 vs 0.021 ± 0.0050 µm 2 / µm 2 adventitial area, p=0.019). Immunodetection of Hypoxyprobe™ showed evidence of increased hypoxia. VVG, pentachrome, and immunodetection of αSMA showed decreased elastin content, increased deposition of ground substance, and areas of smooth muscle cell loss . Multiphoton microscopy demonstrated fragmented elastin fiber microarchitecture and decreased collagen content (Fig 1). Conclusions: Our study findings demonstrate a novel, clinically relevant model for thoracic aortic aneurysm through vasa vasorum inhibition that recapitulates features of cystic medial degeneration. This translational model could enable development of targeted, pro-regenerative therapies that restore perfusion to the aortic wall.

DDDR-25. IBUDILAST AND ANTI-PD-L1 COMBINATION TREATMENT SIGNIFICANTLY IMPROVES SURVIVAL IN A SEX-DEPENDENT MANNER IN A MURINE MODEL OF GLIOBLASTOMA

Abstract INTRODUCTION Glioblastoma (GBM) is the most common primary malignant brain tumor, and there remains a significant need for new treatments. Previous trials studying immune checkpoint inhibitors (ICI) as single agents have been negative. Ibudilast is a brain penetrant inhibitor of the macrophage migration inhibitory factor (MIF)-CD74 interaction that sensitizes GBM to temozolomide in vitro and alters myeloid-derived suppressor cells in vitro and in vivo. The purpose of this study was to investigate whether ibudilast increases ICI efficacy in a murine model of GBM. METHODS 6-week-old male and female C57BL/6 mice were implanted with SB28 cells intracranially. One week after implantation, each group was randomized to receive intraperitoneal treatment with either: ibudilast (50 mg/kg), anti-PD-L1 (1mg/kg), combination, or vehicle control. Mice were euthanized at neurological endpoint. Survival was modeled with Cox proportional hazards regression. RESULTS Among females, there was a significant difference in survival among treatment groups (p=0.0002). Specifically, combination treatment led to significantly longer survival compared to ibudilast (p=0.004) and control (p=0.03), but not anti-PD-L1 (p=0.12). Among males, there was no significant difference in survival among groups (p=0.40). Males had significantly shorter median survival compared to females (overall: 18 versus 22 days, combination treatment: 20 days versus 32 days, anti-PD-L1: 14 versus 22 days, ibudilast: 17 versus 17 days, control: 18 versus 22 days, p=0.02). CONCLUSIONS Ibudilast with anti-PD-L1 is a promising combination treatment against GBM. Future research is needed to identify the mechanisms underlying this treatment regimen and its sex-dependent effect. These results may inform the development of a combination new clinical trial, as ibudilast is currently being evaluated in the newly-diagnosed and recurrent GBM setting with temozolomide.

TMIC-53. USE OF CEREBRAL OPEN-FLOW MICROPERFUSION TO IDENTIFY POTENTIAL ONCOMETABOLITE, L-2-HYDROXYGLUTARATE, IN GLIOBLASTOMA CHEMOTHERAPY ADAPTATION

Abstract Glioblastoma (GBM) is among the most prevalent and lethal brain tumors, comprising nearly half of all central nervous system malignancies and imparting a median overall survival of only 18-20 months. One major challenge in curing this disease is the limited understanding of how GBM cells biologically respond to treatments like radiotherapy and chemotherapy, including temozolomide (TMZ), in real-time. A significant challenge is observing changes within the tumor interstitial fluid (ISF), which mediates adaptive intercellular signaling between tumor cells and their surrounding tumoral and stromal neighbors. Microfluidic sampling methods, such as cerebral microdialysis and microperfusion, have started to provide valuable insights into the ISF signaling dynamics. These methods enable real-time biomolecular monitoring of this often-neglected component of the tumor microenvironment (TME). We have performed in vivo cerebral open-flow microperfusion (cOFM) pilot studies to identify metabolomic and proteomic changes within the tumor ISF resulting from differential treatment with either TMZ or vehicle control. Among these metabolite and protein changes, we identified an increase in 2-hydroxyglutarate (2-HG). While D-2-HG is a hallmark metabolite of isocitrate dehydrogenase mutant (IDH-mut) tumors, our model utilized the IDH wild-type (IDH-WT) CT2A mouse-syngeneic glioma model. Subsequent analysis of enantiomeric confirmation characterized this to be predominantly L-2-HG. Follow-up in vitro studies have shown that elevated L-2-HG plays a functional role in IDH-WT patient-derived glioma cells via non-canonical HIF1α stabilization and subsequent expression of HRE genes. This suggested that L-2-HG may play a similar role in IDH-WT GBM as D-2-HG plays in IDH-mutant tumors. This study demonstrates the potential of L-2-HG to function as a chemo-responsive oncometabolite in the TME of highly aggressive IDH-WT glioblastoma. By targeting the metabolic and signaling properties of L-2-HG, it may be possible to prevent chemo-resistant adaptations mounted by GBM cells when treated with TMZ.

The French customs authority system is an example of effective combatance against customs offenses for Ukraine

The article analyzes the structure and functioning of French customs authorities, emphasizing their role in combating offenses in the customs sphere and the possibility of adapting their experience for Ukraine. The French customs service consists of several units responsible for various aspects of customs control, from intelligence to investigations and technical support. Special attention is paid to the use of innovative technologies, such as risk analysis systems and «smart borders», which significantly increase the efficiency of customs operations. These technologies allow accurate control of goods, vehicles and passengers, which ensures prevention of customs offenses. The results show that the structure of the French customs authorities, divided into specialized directorates (intelligence, operations, investigations and technical support), significantly contributes to the prevention of fraud, illegal trade and security threats. Active use of risk analysis systems and cooperation with international organizations helps France manage cross-border risks and ensure compliance with European standards. The French model offers a valuable plan for the modernization of the customs service of Ukraine and its integration into European and world standards. On the basis of the analysis of the experience of France, conclusions were drawn regarding the possible directions of development of the Ukrainian customs service, in particular through the introduction of modern technologies, automation of processes and strengthening of international cooperation. The integration of advanced risk management methods and automated control systems will allow Ukraine to improve the transparency, security and efficiency of customs operations, reducing the number of offenses and corruption.