OVA-PEG-R848 nanocapsules stimulate neonatal conventional and plasmacytoid dendritic cells

Childhood mortality represents a major issue with 5. 3 million worldwide deaths of children under 5 years of age in 2019. Approximately half of those deaths can be attributed to easily preventable, infectious diseases. Currently approved neonatal vaccines are typically effective only after multiple doses leaving infants especially vulnerable during the first 6 months of life. Survival rates could be improved significantly by developing new and more potent vaccines that are capable of overcoming inherently tolerogenic neonatal immune systems. TLR agonists have garnered a great deal of attention in recent years due to their extensive capacities to activate innate immunity. Herein, the superior capacity of the TLR7/8 agonist, resiquimod (R848), to activate adult and neonatal primary peripheral blood dendritic cells is demonstrated. Moreover, R848 can be conjugated to polyethylene glycol and encapsulated in ovalbumin nanocapsules to efficiently co-deliver antigen and adjuvant in vitro. This study is among the first to demonstrate the capacity of encapsulated R848 to activate neonatal dendritic cells. These findings support the potential incorporation of R848 as adjuvant in neonatal vaccines, making them more effective in eliciting a robust immune response

A-405 - The basal cell carcinoma-one-stop-shop-study [Abstract]

Background: The treatment of basal cell carcinomas (BCCs) poses significant challenges due to the invasive, time-intensive and costly nature of margin mapping techniques such as Mohs micrographic surgery.[1][2] To address these limitations, we developed the "BCC-One-Stop-Shop-Method", integrating in-vivo and ex-vivo margin mapping using Line-field Confocal Optical Coherence Tomography (LC-OCT). This method aims to streamline diagnosis, surgery and margin control process, allowing for completion within one patient visit. This study outlines the process, the results compared to histology and highlights its advantages in clinical practice. Methods: BCC lesions were marked using color-coded tattoo pens to delineate margins in four quadrants. Dermoscopic mosaic images were captured for AI-assisted colocalization, ensuring precise margin mapping. LC-OCT was used in-vivo to record videos along the marked margins with an integrated AI providing real-time BCC probability scores. Images of the center were taken by LC-OCT to assess depth and subtype. After excision, the tissue was placed in an ex-vivo setup, with the LC-OCT handheld adapted to a movable sample holder for imaging of the specimen. Videos and 3D stacks were recorded to confirm margin clearance. Then the tissue undergoes conventional histological processing for correlation with LC-OCT findings. Results: 50 BCCs of 43 patients were examined, 32 lesions also with ex-vivo LC-OCT. 195 tissue quarters were evaluated and compared. 38 lesions were located on the head (76 %), 7 on the trunk (14 %), 5 on the extremities (10 %). 6 BCCs were superficial, 26 nodular, 14 infiltrative, 4 mixed subtypes and 1 was undefined. For in-vivo LC-OCT vs. histology sensitivity was 81.8 % (86.4 %), specificity 94.8 % (96.5 %) for the lesion (quarter) level. For ex-vivo LC-OCT vs. histology sensitivity was 71.4 % (86.6 %), specificity 96.0 % (96.4 %) for the lesion (quarter) level. Accuracy of overall performances for in-vivo for the lesion (quarter) level was 92 % (95.4 %), for ex-vivo 90.6 % (81.1 %). Conclusions: Our study presents a new approach for integrating advanced imaging technologies into BCC management, offering a comprehensive solution for preoperative margin mapping and intraoperative evaluation. Initial findings suggest this method is efficient, easy to implement, and well-accepted by patients. Further studies will confirm its efficacy and potential in BCC treatment, potentially reducing the psychological and economic burden associated with traditional methods

Controlling protein interactions in blood for effective liver immunosuppressive therapy by silica nanocapsules

Immunosuppression with glucocorticoids is a common treatment for autoimmune liver diseases and after liver transplant, which is however associated with severe side-effects. Targeted delivery of glucocorticoids to inflammatory cells, e.g. liver macrophages and Kupffer cells, is a promising approach for minimizing side effects. Herein, we prepare core–shell silica nanocapsules (SiO2 NCs) via a sol–gel process confined in nanodroplets for targeted delivery of dexamethasone (DXM) for liver immunosuppressive therapy. DXM with concentrations up to 100 mg mL−1 in olive oil are encapsulated while encapsulation efficiency remains over 95% after 15 days. Internalization of NCs by non-parenchymal murine liver cells significantly reduces the release of inflammatory cytokines, indicating an effective suppression of inflammatory response of liver macrophages. Fluorescent and magnetic labeling of the NCs allows for monitoring their intracellular trafficking and biodegradation. Controlled interaction with blood proteins and good colloidal stability in blood plasma are achieved via PEGylation of the NCs. Specific proteins responsible for stealth effect, such as apolipoprotein A-I, apolipoprotein A-IV, and clusterin, are present in large amounts on the PEGylated NCs. In vivo biodistribution investigations prove an efficient accumulation of NCs in the liver, underlining the suitability of the SiO2 NCs as a dexamethasone carrier for treating inflammatory liver diseases.Fil: Jiang, Shuai. Max-Planck-Institut für Polymerforschung; AlemaniaFil: Prozeller, Domenik. Max-Planck-Institut für Polymerforschung; AlemaniaFil: Pereira, Jorge. Max-Planck-Institut für Polymerforschung; AlemaniaFil: Simon, Johanna. Max-Planck-Institut für Polymerforschung; Alemania. Johannes Gutenberg Universitat Mainz; AlemaniaFil: Han, Shen. Max-Planck-Institut für Polymerforschung; AlemaniaFil: Wirsching, Sebastian. Johannes Gutenberg Universitat Mainz; AlemaniaFil: Fichter, Michael. Johannes Gutenberg Universitat Mainz; AlemaniaFil: Mottola, Milagro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina. Max-Planck-Institut für Polymerforschung; AlemaniaFil: Lieberwirth, Ingo. Max-Planck-Institut für Polymerforschung; AlemaniaFil: Morsbach, Svenja. Max-Planck-Institut für Polymerforschung; AlemaniaFil: Mailänder, Volker. Max-Planck-Institut für Polymerforschung; Alemania. Johannes Gutenberg Universitat Mainz; AlemaniaFil: Gehring, Stephan. Johannes Gutenberg Universitat Mainz; AlemaniaFil: Crespy, Daniel. Max-Planck-Institut für Polymerforschung; Alemania. Vidyasirimedhi Institute of Science and Technology; TailandiaFil: Landfester, Katharina. Max-Planck-Institut für Polymerforschung; Alemani

Supramolecular Toxin Complexes for Targeted Pharmacological Modulation of Polymorphonuclear Leukocyte Functions

The targeted pharmacological modulation of polymorphonuclear leukocytes (PMNs) is of major medical interest. These innate immune cells play a central role in the defense against pathogenic microorganisms. However, their excessive chemotactic recruitment into tissues after traumatic injury is detrimental due to local and systemic inflammation. Rho-GTPases, being the master regulators of the actin cytoskeleton, regulate migration and chemotaxis of PMNs, are attractive pharmacological targets. Herein, supramolecular protein complexes are assembled in a “mix-and-match” approach containing the specific Rho-inhibiting clostridial C3 enzyme and three PMN-binding peptides using an avidin platform. Selective delivery of the C3 Rho-inhibitor with these complexes into the cytosol of human neutrophil-like NB-4 cells and primary human PMNs ex vivo is demonstrated, where they catalyze the adenosine diphosphate (ADP) ribosylation of Rho and induce a characteristic change in cell morphology. Notably, the complexes do not deliver C3 enzyme into human lung epithelial cells, A549 lung cancer cells, and immortalized human alveolar epithelial cells (hAELVi), demonstrating their cell type-selectivity. The supramolecular complexes represent attractive molecular tools to decipher the role of PMNs in infection and inflammation or for the development of novel therapeutic approaches for diseases that are associated with hyperactivity and reactivity of PMNs such as post-traumatic injury

Integrated Molecular-Morphologic Meningioma Classification: A Multicenter Retrospective Analysis, Retrospectively and Prospectively Validated

PURPOSE: Meningiomas are the most frequent primary intracranial tumors. Patient outcome varies widely from benign to highly aggressive, ultimately fatal courses. Reliable identification of risk of progression for individual patients is of pivotal importance. However, only biomarkers for highly aggressive tumors are established (CDKN2A/B and TERT), whereas no molecularly based stratification exists for the broad spectrum of patients with low- and intermediate-risk meningioma. METHODS: DNA methylation data and copy-number information were generated for 3,031 meningiomas (2,868 patients), and mutation data for 858 samples. DNA methylation subgroups, copy-number variations (CNVs), mutations, and WHO grading were analyzed. Prediction power for outcome was assessed in a retrospective cohort of 514 patients, validated on a retrospective cohort of 184, and on a prospective cohort of 287 multicenter cases. RESULTS: Both CNV- and methylation family-based subgrouping independently resulted in increased prediction accuracy of risk of recurrence compared with the WHO classification (c-indexes WHO 2016, CNV, and methylation family 0.699, 0.706, and 0.721, respectively). Merging all risk stratification approaches into an integrated molecular-morphologic score resulted in further substantial increase in accuracy (c-index 0.744). This integrated score consistently provided superior accuracy in all three cohorts, significantly outperforming WHO grading (c-index difference P = .005). Besides the overall stratification advantage, the integrated score separates more precisely for risk of progression at the diagnostically challenging interface of WHO grade 1 and grade 2 tumors (hazard ratio 4.34 [2.48-7.57] and 3.34 [1.28-8.72] retrospective and prospective validation cohorts, respectively). CONCLUSION: Merging these layers of histologic and molecular data into an integrated, three-tiered score significantly improves the precision in meningioma stratification. Implementation into diagnostic routine informs clinical decision making for patients with meningioma on the basis of robust outcome prediction

A Damage Mechanics Approach to Fatigue Assessment in Offshore Structures

This article is intended to describe the development of a fatigue damage model capable of assessing fatigue damage in offshore structures. This is achieved by for mulating a set of damage coupled constitutive and evolution equations which make the for mulation of a unified approach possible under both low and high cycle fatigue damage and consistent with the structural dynamic response of the changing/deteriorating material be haviors. The structural analysis for the whole designed period, say about 30 years, can be carried out with the aid of the proposed analytical procedure, in which the fundamental characteristics of sea wave statistics responsible for the structural dynamic response can be sufficiently considered. An offshore structure subject to complex ocean environment is described by a general stochastic system which embeds a group of stochastic subsystems, each characterizing a duty cycle. An effective analytical method is established by introduc ing the concept of duty strain range with a clear mathematical definition and its analytical solution which covers all possible spectral parameters. The history-dependent damage is also included in the damage model so that the overload effects can be analyzed. It should be pointed out that the whole procedure can be fully computerized such that the practical or engineering significance of varying design variables can be readily highlighted.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/67255/2/10.1177_105678959300200405.pd

A T-cell antigen atlas for meningioma: novel options for immunotherapy

Meningiomas are the most common primary intracranial tumors. Although most symptomatic cases can be managed by surgery and/or radiotherapy, a relevant number of patients experience an unfavorable clinical course and additional treatment options are needed. As meningiomas are often perfused by dural branches of the external carotid artery, which is located outside the blood-brain barrier, they might be an accessible target for immunotherapy. However, the landscape of naturally presented tumor antigens in meningioma is unknown. We here provide a T-cell antigen atlas for meningioma by in-depth profiling of the naturally presented immunopeptidome using LC-MS/MS. Candidate target antigens were selected based on a comparative approach using an extensive immunopeptidome data set of normal tissues. Meningioma-exclusive antigens for HLA class I and II are described here for the first time. Top-ranking targets were further functionally characterized by showing their immunogenicity through in vitro T-cell priming assays. Thus, we provide an atlas of meningioma T-cell antigens which will be publicly available for further research. In addition, we have identified novel actionable targets that warrant further investigation as an immunotherapy option for meningioma