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Advances in RNA interference technology have established it as a powerful therapeutic tool with important future potential. The design and the chemical modifications of the siRNA nucleotide backbone have greatly enhanced stability, durability, and pharmacokinetics while minimizing tolerability risks. The optimal combination of these modifications depends on the target gene, tissue, and RNA sequence, necessitating an iterative, experimental approach that currently relies heavily on animal models. To reduce the reliance and number of (humanized) animals required, we developed a human long‐term liver 3D spheroid model designed for screening GalNAc‐conjugated siRNAs which captures the process of uptake, potency, and durability for early in vitro screening. These liver spheroids remain viable in culture for at least 5 weeks while maintaining expression of the asialoglycoprotein receptor to facilitate GalNAc mediated uptake. siRNA was efficiently internalized by the spheroids without the need for transfection reagents, and its durable silencing efficiency was assessed by monitoring AHSA1 target gene expression over time. Target gene silencing in the spheroid model persisted up to 5 weeks post‐treatment. Fluorescently labeled siRNA enabled visualization of uptake and distribution within the spheroid, revealing somewhat reduced siRNA accumulation in pericentral CYP3A4+ hepatocytes accompanied with somewhat reduced ASGR1 expression. No signs of hepatotoxicity were observed under the conditions used. By varying the number of phosphorothioate modifications in the siRNA backbone, distinct differences in silencing efficiency and durability were observed which were principally similar as obtained in vivo in mice. We propose that this long‐term human liver spheroid model provides a valuable preclinical platform for evaluating siRNA‐based therapeutics with respect to uptake, durability, and silencing efficiency, and could refine early in vitro screening and accelerate drug development. Study Highlights What is the current knowledge on the topic? ○Chemical modifications to the siRNA backbone are essential for enhancing molecular stability, reducing immunogenicity, increasing resistance to enzymatic degradation, and minimizing toxicity. ○Such modifications are pivotal for optimizing siRNA performance in therapeutic settings. ○However, despite growing interest in siRNA‐based therapeutics, no long‐term in vitro model exists to evaluate silencing efficiency and durability. ○As a result, preclinical development still relies heavily on humanized mouse models. What question did this study address? ○Can the human liver spheroids be used as an in vitro model to evaluate the effect of backbone modifications of the GalNAc‐conjugated siRNA on the silencing efficiency and durability? What does this study add to our knowledge? ○We present a new human spheroid model for assignment of the properties of structurally different GalNAc‐conjugated siRNA molecules. ○The spheroids were maintained in culture for up to 5 weeks while retaining expression of the asialoglycoprotein receptor. ○These spheroids can be directly treated with GalNAc‐conjugated siRNA without the need for transfection reagents, enabling sustained target gene silencing to be monitored over the entire five‐week period. ○Using fluorescently labeled siRNA, intracellular trafficking was visualized, revealing heterogeneity in hepatocyte uptake capacity. ○Moreover, the model enabled clear discrimination of silencing performance across five distinct siRNA backbones. How might this change clinical pharmacology or translational science? ○This study shows that human liver spheroids provide a robust, scalable platform for evaluating GalNAc‐conjugated siRNA therapeutics. ○The model offers a valuable tool for early‐stage compound selection, potentially accelerating screening workflows and reducing reliance on animal models in preclinical development.

Antibody drug conjugates (ADC) are a promising class of oncology therapeutics consisting of an antibody conjugated to a payload via a linker. DYP688 is a novel ADC comprising of a signaling protein inhibitor payload (FR900359) that undergoes unique on-antibody inactivation in plasma, resulting in complex pharmacology. To assess the impact of FR inactivation on DYP688 pharmacology and clinical developability, we performed translational modeling of preclinical PK and tumor growth inhibition (TGI) data, accompanied by mechanistic Krogh cylinder tumor modeling. Using a PK-TGI model, we identified a composite exposure-above-tumorostatic concentration (AUC TSC ) metric as the PK-driver of efficacy. To underpin the mechanisms behind AUC TSC as the driver of efficacy, we performed quantitative systems pharmacology (QSP) modeling of DYP688 intratumoral pharmacokinetics and pharmacodynamics. Through exploratory simulations, we show that by deviating from canonical ADC design dogma, DYP688 has optimal FR900359 activity despite its transient inactivation. Finally, we performed the successful preclinical to clinical translation of DYP688 PK, including the payload inactivation kinetics, evidenced by good agreement of the predicted PK to the observed interim clinical PK. Overall, this work highlights early quantitative pharmacokinetics as a missing link in the ADC design-developability chasm.

The performance of low-power, continuous-wave stimulated emission depletion microscopy is improved by combining pulsed excitation with time-gated detection. This combination also simplifies super-resolution fluorescence correlation spectroscopy. Applying pulsed excitation together with time-gated detection improves the fluorescence on-off contrast in continuous-wave stimulated emission depletion (CW-STED) microscopy, thus revealing finer details in fixed and living cells using moderate light intensities. This method also enables super-resolution fluorescence correlation spectroscopy with CW-STED beams, as demonstrated by quantifying the dynamics of labeled lipid molecules in the plasma membrane of living cells.

‘Pope calls on G7 leaders to ban use of autonomous weapons’ (The Guardian 2024) is the headline from statements which pope Franziskus made during the G7-summit on June 14th, 2024. In general it can be observed that the ethical debate concerning the use of autonomous weapon systems (AWS) is an extremely complex and contentious issue, raising both technical and ethical challenges. Through a comparative analysis of relevant literature the theological perspective is introduced into the debate and highlight potential implications for the use of AWS. The increasing autonomy, where machines can autonomously select and engage targets, raises questions regarding compliance with international humanitarian law, the preservation of human dignity and moral responsibility. The research question addressed in this article is as follows: ‘What are the theological-ethical arguments regarding the use of AWS?’ In conclusion, three key points for a theological-ethical examination consist of the question about the pessimistic human image as a premise of the pro-AWS argumentation and ethical questions based on the highest or preferable good as well as on moral responsibility. It is synthesised, that the pessimistic human image can be represented, that as highest good right to life should be preferred against human dignity and that moral responsibility always should stick on humans and not on AWS.Intradisciplinary and/or interdisciplinary implicationsThis article positions theological ethics within the emerging field of ethical dilemmas arising from autonomous functions in the realm of technical ethics.

Pulmonary infections are characterized by neutrophil recruitment into the lung driven by chemokine ligands of CXCR2, which is expressed on neutrophils, but also present in non-hematopoietic lung cells, in which its role remains unclear. We hypothesize that CXCR2 in epithelial and endothelial cells contributes to neutrophil recruitment into the lung by modifying the availability of its cognate chemokines in lung alveoli. Using conditional endothelial and epithelial CXCR2 knockout mice, we demonstrate that selective CXCR2 deletion in either compartment impairs neutrophil recruitment into the lung during bacterial pneumonia and reduces bacterial clearance. We show that CXCR2 ablation in epithelial and endothelial cells compromises respective trans-epithelial and trans-endothelial transcytosis of alveolar CXCL1. Mechanistically, CXCR2-mediated CXCL1 endothelial and epithelial cell transcytosis requires the function of Bruton’s tyrosine kinase in these cells. In conclusion, CXCR2 plays an important role in alveolar epithelial and endothelial cells, where it mediates cognate chemokine transcytosis, thus actively supporting their activities in neutrophil recruitment to the infected lungs. The role of CXCR2 in epithelial and endothelial cells on lung infection remains unclear. The authors here use conditional CXCR2 knockout mice to manifest that epithelial and vascular CXCR2 mediates transcytosis of CXCL1, leading to neutrophil infiltration and controlled lung inflammation.

CKAMP44, identified here by a proteomic approach, is a brain-specific type I transmembrane protein that associates with AMPA receptors in synaptic spines. CKAMP44 expressed in Xenopus oocytes reduced GluA1- and A2-mediated steady-state currents, but did not affect kainate- or N-methyl- D-aspartate (NMDA) receptor-mediated currents. Mouse hippocampal CA1 pyramidal neurons expressed CKAMP44 at low abundance, and overexpression of CKAMP44 led to stronger and faster AMPA receptor desensitization, slower recovery from desensitization, and a reduction in the paired-pulse ratio of AMPA currents. By contrast, dentate gyrus granule cells exhibited strong CKAMP44 expression, and CKAMP44 knockout increased the paired-pulse ratio of AMPA currents in lateral and medial perforant path-granule cell synapses. CKAMP44 thus modulates short-term plasticity at specific excitatory synapses.

In this contribution, the first amidinate and amidine derivatives of p-carborane are described. Double lithiation of p-carborane (1) with n-butyllithium followed by treatment with 1,3-diorganocarbodiimides, R–N=C=N–R (R = iPr, Cy (= cyclohexyl)), in DME or THF afforded the new p-carboranylamidinate salts p-C2H10B10[C(NiPr)2Li(DME)]2 (2) and p-C2H10B10[C(NCy)2Li(THF)2]2 (3). Subsequent treatment of 2 and 3 with 2 equiv. of chlorotrimethylsilane (Me3SiCl) provided the silylated neutral bis(amidine) derivatives p-C2H10B10[CiPrN(SiMe3)(=NiPr)]2 (4) and p-C2H10B10[CCyN(SiMe3)(=NCy)]2 (5). The new compounds 3 and 4 have been structurally characterized by single-crystal X-ray diffraction. The lithium carboranylamidinate 3 comprises a rare trigonal planar coordination geometry around the lithium ions.

Environmental stress like important soccer events can induce excitation, stress and anger. We aimed to investigate (i) whether the FIFA soccer world cup (WC) 2014 and (ii) whether the soccer games of the German national team had an impact on total numbers and in-hospital mortality of patients with myocardial infarction (MI) in Germany. We analyzed data of MI inpatients of the German nationwide inpatient sample (2013–2015). Patients admitted due to MI during FIFA WC 2014 (12th June–13th July2014) were compared to those during the same period 2013 and 2015 (12th June–13th July). Total number of MI patients was higher during WC 2014 than in the comparison-period 2013 (18,479 vs.18,089, P  < 0.001) and 2015 (18,479 vs.17,794, P  < 0.001). WC was independently associated with higher MI numbers (2014 vs. 2013: OR 1.04 [95% CI 1.01–1.07]; 2014 vs. 2015: OR 1.07 [95% CI 1.04–1.10], P  < 0.001). Patient characteristics and in-hospital mortality rate (8.3% vs. 8.3% vs. 8.4%) were similar during periods. In-hospital mortality rate was not affected by games of the German national team (8.9% vs. 8.1%, P  = 0.110). However, we observed an increase regarding in-hospital mortality from 7.9 to 9.3% before to 12.0% at final-match-day. Number of hospital admissions due to MI in Germany was 3.7% higher during WC 2014 than during the same 31-day period 2015. While in-hospital mortality was not affected by the WC, the in-hospital mortality was highest at WC final.

In this contribution, the first amidinate and amidine derivatives of p-carborane are described. Double lithiation of p-carborane (1) with n-butyllithium followed by treatment with 1,3-diorganocarbodiimides, R-N=C=N-R (R = iPr, Cy (= cyclohexyl)), in DME or THF afforded the new p-carboranylamidinate salts p-C[sub.2]H[sub.10]B[sub.10][C(NiPr)[sub.2]Li(DME)][sub.2] (2) and p-C[sub.2]H[sub.10]B[sub.10][C(NCy)[sub.2]Li(THF)[sub.2]][sub.2] (3). Subsequent treatment of 2 and 3 with 2 equiv. of chlorotrimethylsilane (Me[sub.3]SiCl) provided the silylated neutral bis(amidine) derivatives p-C[sub.2]H[sub.10]B[sub.10][CiPrN(SiMe[sub.3])(=NiPr)][sub.2] (4) and p-C[sub.2]H[sub.10]B[sub.10][CCyN(SiMe[sub.3])(=NCy)][sub.2] (5). The new compounds 3 and 4 have been structurally characterized by single-crystal X-ray diffraction. The lithium carboranylamidinate 3 comprises a rare trigonal planar coordination geometry around the lithium ions.

Background and aimsWith an external additional working channel (AWC) endoscopic mucosal resection (EMR) as well as endoscopic submucosal dissection (ESD) can be extended to techniques termed “EMR+” and “ESD+.” These novel techniques are systematically compared to EMR and ESD under the use of a double-channel endoscope (DC).MethodsOur trial was conducted prospectively in a pre-clinical porcine animal model (EASIE-R simulator) with standardized gastric lesions measuring 3 or 4 cm.ResultsEMR+ and EMR DC showed both good results for 3 cm lesions with no adverse events and an en bloc resection rate of 73.33% (EMR+) and 60.00% (EMR DC, p = 0.70). They came to their limits in 4 cm lesions with muscularis damages of 20.00% (EMR+), 13.33% (EMR DC, p ≥ 0.99) and decreasing en bloc resection rates of 60.00% (EMR+) and 46.67% (EMR DC, p = 0.72).ESD+ and ESD DC were both reliable concerning en bloc resection rates (100% in all groups) and adverse events (0.00% in 3 cm lesions, 12.50% muscularis damages in both ESD+ and ESD DC in 4 cm lesions).Resection time was slightly shorter in all groups with the AWC compared to DC although only reaching significance in 3 cm ESD lesions (p < 0.05*).ConclusionsWith the AWC, a standard endoscope can easily be transformed to double-channel functionality. We could show that EMR+ and ESD+ are non-inferior to EMR and ESD under the use of a double-channel endoscope. Consequently, the AWC presents an affordable alternative to a double-channel endoscope for both EMR and ESD.