University Hospital Frankfurt

Hirschsprung disease (HD) is a congenital disorder marked by aganglionosis of intestinal nerve plexuses, leading to bowel obstruction. Determining the extent of the aganglionic segment is essential for successful surgical correction. Consequently, the extent of aganglionosis must be examined histologically. Fluorescence confocal microscopy (FCM) may serve as an alternative diagnostic modality. Using a laser to scan the unfixed specimen, it enables rapid intraoperative imaging without causing tissue alteration. This study assessed the feasibility and diagnostic accuracy of FCM in evaluating Hirschsprung disease in pediatric patients. Patients undergoing rectal biopsy, ostomy closure, or transanal endorectal pull-through (TERPT) at our center from January to August 2024 were included. Tissue samples were imaged by FCM followed by frozen section and formalin-fixed paraffin-embedded (FFPE) histology. Image quality, assessability of submucosal and myenteric plexuses, and diagnostic agreement with FFPE sections were investigated. A total of 34 samples from 8 patients were analyzed by FCM and FFPE, the 24 TERPT-derived samples were additionally analyzed by frozen section. Diagnostic agreement with FFPE was 78.5 % for FCM and 86.9 % for frozen sections (p = 0.49). Among fully assessable samples, FCM achieved 80.9 % agreement with FFPE compared to 86.4 % for frozen sections. FCM is an easy-to-implement method for identifying normal and affected bowel in HD. It is used on unfixed specimens, preserving the tissue quality. Its diagnostic accuracy is comparable to that of frozen sections in assessable samples, and it is advantageous in cases where local access to expert pathologists is limited.

Hereditary angioedema is a bradykinin-mediated, rare condition characterised by recurrent and potentially life-threatening attacks of subcutaneous and submucosal swelling. Bradykinin B2 receptor antagonism is a proven mechanism for on-demand treatment of attacks, but no evidence exists on its effects when used prophylactically. Deucrictibant is an investigational orally bioavailable bradykinin B2 receptor antagonist. We aimed to evaluate the efficacy, safety, and tolerability of two dose regimens of oral deucrictibant administered as prophylaxis against hereditary angioedema attacks. CHAPTER-1 was a multicentre, double-blind, placebo-controlled, randomised, phase 2 trial conducted in two parts, a double-blind placebo controlled first part and an open-label second part, with only part 1 reported here. Part 1 recruited adults (aged 18-75 years) with hereditary angioedema type 1 or 2 from 37 sites (university hospitals and accredited angioedema centres) across North America, Europe, and Israel. Patients required a documented history of three or more attacks within the last 3 consecutive months before screening or two or more during the screening period (up to 8 weeks) to be eligible. An interactive response technology system randomised eligible patients 1:1:1 to receive oral deucrictibant 20 mg daily, 40 mg daily, or matching placebo for 12 weeks. Randomisation to treatment groups was stratified by the baseline attack rate. Patients, investigators, site personnel, and the sponsor were blinded to treatment assignment. Masking was achieved with identically appearing deucrictibant and placebo capsules. The primary endpoint was the time-normalised number of investigator-confirmed attacks per 4 weeks (monthly attack rate) from weeks 1 to 12 and was assessed using the intention-to-treat set. The endpoint was analysed by comparing each deucrictibant group with the placebo group using a Poisson generalised linear model with a log link function and Pearson's χ2 scaling of SEs to account for potential dispersion. The safety analysis set included all patients who were randomly assigned and who received one or more doses of study drug (deucrictibant or placebo). CHAPTER-1 is registered with ClinicalTrials.gov (NCT05047185) and is now complete. Between March 9, 2022, and June 19, 2023, 44 patients were screened. Of 34 patients who were randomly assigned, 11 patients received deucrictibant 20 mg, 12 patients received deucrictibant 40 mg, and 11 patients received the placebo, with a median follow-up of 85·0 days (IQR 84·0-86·0). 21 (62%) patients were female, 13 (38%) were male, and 34 (100%) patients were White. The least squares mean monthly attack rate (primary analysis) was 0·40 (95% CI 0·18-0·92) for deucrictibant 20 mg, 0·30 (0·11-0·81) for deucrictibant 40 mg, and 1·93 (1·30-2·88) for placebo; percent reduction in attack rate compared with placebo was 79·2% (95% CI 47·2-91·8) for deucrictibant 20 mg (p=0·0010) and 84·5% (95% CI 53·8-94·8) for deucrictibant 40 mg (p=0·0008). Treatment-related treatment-emergent adverse events were experienced by two (18%) patients receiving deucrictibant 20 mg, one (8%) patient receiving deucrictibant 40 mg, and one (9%) patient receiving the placebo; all were mild in severity (grade 1) and did not require dosing modification of the study drug. There were no serious adverse events or deaths in any treatment group. To the best of our knowledge, this trial provides the first clinical evidence and proof-of-concept for bradykinin B2 receptor antagonism as a therapeutic approach for the prevention of hereditary angioedema attacks and supports further investigation of oral deucrictibant for bradykinin-mediated angioedema. Pharvaris.

The skin is a diverse reservoir of immune cells with strong potential for immunotherapeutic delivery. Langerhans cells in the epidermis are antigen-presenting cells, accessible for vaccination using carbohydrate-conjugated therapeutics targeting their endocytic lectin receptor, Langerin. As carbohydrate-lectin binding is highly dependent on valency, scaffolds that enable control over ligand spacing and stoichiometry are instrumental in enhancing receptor binding and selectivity. Here we utilized a self-assembled nucleic acid-based Holliday Junction scaffold, fully modified for nuclease protection and with a well-defined carbohydrate arrangement to optimize drug delivery to Langerhans cells. In vitro screening with Langerin-expressing cells revealed that mannosylated Holliday Junctions showed the strongest binding. This was confirmed in human epidermal cell suspensions, demonstrating specificity and valency-driven interactions. Topical administration of mannosylated scaffolds on skin explants enabled effective targeting of epidermal Langerhans cells. Finally, in an antigen-presentation assay, in vitro differentiated Langerhans cells loaded with mannosylated and peptide-conjugated scaffolds significantly enhanced T cell activation. Overall, our study presents a promising nucleic acid-based platform for precise Langerhans cell targeting and drug delivery, with broad potential for skin-directed immunotherapies.

The study we describe focuses on evaluating the effectiveness of linear accelerator (LINAC) stereotactic radiosurgery (SRS) in the treatment of cerebral arteriovenous malformations (AVMs). This treatment option is gaining interest due to the uncertainties associated with combined radiosurgical and endovascular treatments and the significant technological advancements in SRS. The primary goals of the study are to assess rates of obliteration (successful closure of the AVM) and rebleeding (the recurrence of bleeding posttreatment), as well as to identify factors influencing obliteration rates and to document any adverse effects associated with the procedure. The study retrospectively analyzed data from 134 patients treated with LINAC-based SRS for cerebral AVMs. The patients were categorized based on their prior treatments: 50had undergone partial embolization, 8had received acombination of embolization and surgery and 1patient had asurgical intervention. Furthermore, 75patients had received no prior treatment. Kaplan-Meier survival analysis and log-rank tests were employed to calculate actuarial obliteration rates and annual cumulative bleeding rates following SRS treatment. The study found that obliteration rates after SRS treatment increased over time, with 5‑year obliteration rates of 85.2% for grades I-II, 76.4% for gradeIII, and 62.1% for grades IV-V. Annual cumulative bleeding rates post-SRS were 1.5% for the first year and 0.7% for the second year. Interestingly, prior embolization did not affect the obliteration rate. The median time to obliteration was 36months (range 7-162 months). Obliteration rates were significantly better in Spetzler-Martin (SM) gradeI-II (85% at 5years) compared to gradeIII-V (68% at 5years, p = 0.01). Age, sex, and pediatric status had no statistically significant influence on AVM response to SRS. No radiation necrosis was observed in our cohort. This study contributes to the body of evidence supporting the effectiveness of SRS in treating cerebral AVMs and provides valuable insights into factors affecting treatment outcomes. The findings suggest that patients can expect ahigh chance of successful obliteration of the AVM with minimal adverse effects, making SRS acompelling option for those affected by this condition.

Stereotactic radiotherapy (SRT) is astandard treatment for vestibular schwannoma (VS). Long-term comparative data on single-fraction radiosurgery (SRS) versus normofractionated SRT (NFSRT) are limited. We retrospectively analyzed 175 VS patients treated between 1998 and 2023 (SRS n = 69; NFSRT n = 106; median age61years; median follow-up 46months). Tumor control was evaluated with Kaplan-Meier and Cox regression. Group differences were tested with Mann-WhitneyU and chi-square/Fisher's exact tests. Symptom changes were assessed with McNemar tests, and binary logistic regression identified predictors of acute and late toxicity. Overall, in-field PFS was 94.3% and out-field was PFS was 99.4%. All 10recurrences (median 58.5months) occurred in NFSRT patients treated with 55.8-56 Gy. Higher total dose predicted recurrence (HR = 2.97; p = 0.003). At baseline, 97.1% reported symptoms (hearing loss 86.9%, vertigo 48.0%, tinnitus 42.3%). After therapy, symptoms remained stable, except the incidence of headache, which increased from 14.3% to 22.3% (p = 0.02). Early toxicities were more common after NFSRT, including headache (OR = 4.05; p = 0.01), fatigue (OR = 5.12; p = 0.01), and alopecia (OR = 19.9; p = 0.04), but had resolved by late follow-up. Age and prior surgery predicted vertigo patterns over time. Radionecrosis was rare (0.6%). Both SRS and NFSRT achieve excellent long-term tumor control with low severe toxicity. For NFSRT, normofractionated doses above 55 Gy did not improve efficacy and may increase the recurrence risk, supporting moderate-dose regimens (50-54 Gy). Symptom stabilization confirms the safety of SRT, though headache and vertigo remain relevant long-term issues. Careful dose selection, patient counseling, and rehabilitation are essential to optimize outcomes.

Standard of care for non-metastatic colon cancer is surgery followed by stage-guided adjuvant therapy and/or structured follow-up. Upfront surgery in resectable colon cancer is irrespective of local T/N stage, whereas adjuvant systemic therapy is recommended according to pathological staging. The role of neoadjuvant chemotherapy remains unclear. Whereas perioperative systemic therapy in colon cancer seems to be safe and may lead to pathologic downstaging, evidence on improved survival and quality of life remains scarce. The PROTECTOR / FIRE‑10 trial aims to generate evidence that perioperative systemic therapy improves survival without compromising quality of life in locally advanced, mismatch-repair proficient colon cancer patients. Open-label, randomized, controlled, multicenter, phase III study with two parallel arms. Patients with locally advanced colon or upper rectal cancer staged cT3-4 and/or cN+ are randomized in a 2:1 fashion (favoring preoperative therapy) to investigate the efficacy, patient reported quality of life, and safety of preoperative therapy followed by surgery (Arm A) versus direct surgery followed by non-study specific stage-guided adjuvant therapy (Arm B). Stratification during randomization will be performed according to the following parameters: Fit for mFOLFOXIRI vs. mFOLFOX/CAPOX vs. 80%-mFOLFOX/CAPOX, ECOG 0 vs. ECOG 1-2, and left-sided primary vs. right-sided primary tumor. Only patients with confirmed mismatch-repair proficient and/or microsatellite stable tumor can be included. Preoperative treatment in Arm A is performed for a maximum of 6 biweekly cycles of FOLFOX/FOLFOXIRI or for a maximum of 4 triweekly cycles CAPOX (i.e., appr. 12 weeks). Patients in both arms should undergo quality-controlled surgery of the primary tumor, performed as complete mesocolic excision. Patients will be followed up with regard to relapse, survival and if applicable subsequent anti-cancer treatments until death or for at least 5 years after randomization, whichever date is earlier. The PROTECTOR / FIRE‑10 trial compares preoperative systemic therapy to upfront surgery (with stage-guided adjuvant therapy) in patients with locally advanced colon cancer. This study is registered with clinicaltrials.gov (NCT06899477) and EudraCT (2023-508076-11-00).

IntroductionSézary syndrome (SS) is a rare neoplasm associated with poor prognosis. Due to the rarity of the disease, large-scale investigations are scarce, while identifying prognostic factors is essential for improving treatment outcomes. The primary objective was to investigate prognostic factors in a large population of SS patients on survival and infections. The secondary objective was to validate a previously suggested prognostic index model.MethodsWe conducted a propensity score-matched retrospective cohort study using real-world data from 1277 patients in the TriNetX database. Risk factors examined included age, elevated lactate dehydrogenase (LDH), anemia, elevated white blood cell count (WBC), sex, and race. Examined outcomes were: 5-year survival, Systemic Inflammatory Response Syndrome (SIRS), sepsis and pneumonia. The prognostic index model was simplified into factors assessable through TriNetX.ResultsAge > 60 years, elevated LDH, anemia and elevated WBC were risk factors for 5-year overall survival. Elevated LDH, anemia, elevated WBC and African American/black race were significantly associated with sepsis. Anemia and elevated WBC were also linked to higher risk of pneumonia. Additionally, we devised and validated a simplified prognostic index model (sPIM) accounting for patient focus on SS (stage IV) patients.ConclusionIn the largest group of SS patients assessed for risk stratification to-date, we confirm age and elevated LDH as risk factors in SS and their applicability in the prognostic index model. This study introduces anemia as a novel risk factor and demonstrates the prognostic value of leukocytosis in SS. While race requires further investigation, sex appears to have no significant impact on survival.

Optical calcium imaging is a powerful tool for recording neural activity across a wide range of spatial scales, from dendrites and spines to whole-brain imaging through two-photon and widefield microscopy. Traditional methods for analyzing functional calcium imaging data rely heavily on spatial features, such as the compact shapes of somas, to extract regions of interest and their associated temporal traces. This spatial dependency can introduce biases in time trace estimation and limit the applicability of these methods across different neuronal morphologies and imaging scales. To address these limitations, the Graph Filtered Temporal Dictionary Learning (GraFT) uses a graph-based approach to identify neural components based on shared temporal activity rather than spatial proximity, enhancing generalizability across diverse datasets. Here we present significant advancements to the GraFT algorithm, including the integration of a more efficient solver for the L1 least absolute shrinkage and selection operator (LASSO) problem and the application of compressive sensing techniques to reduce computational complexity. By employing random projections to reduce data dimensionality, we achieve substantial speedups while maintaining analytical accuracy. These advancements significantly accelerate the GraFT algorithm, making it more scalable for larger and more complex datasets. Moreover, to increase accessibility, we developed a graphical user interface to facilitate running and analyzing the outputs of GraFT. Finally, we demonstrate the utility of GraFT to imaging data beyond meso-scale imaging, including vascular and axonal imaging.