Endpoint Measures Research Articles

Prospective Multicenter Evaluation of the MDS "Suggestive of PSP" Diagnostic Criteria.

The recent Movement Disorders Society (MDS)-progressive supranuclear palsy (PSP) diagnostic criteria conceptualized three clinical diagnostic certainty levels: "suggestive of PSP" for sensitive early diagnosis based on subtle clinical signs, "possible PSP" balancing sensitivity and specificity, and "probable PSP" highly specific for PSP pathology. The aim of this study was to prospectively validate the criteria against long-term clinical follow-up and characterize the diagnostic certainty increase over time. Patients with "possible PSP" or "suggestive of PSP" diagnosis and clinical follow-up were recruited in two German multicenter longitudinal observational studies (ProPSP and DescribePSP). The cumulative percentage of patients longitudinally increasing diagnostic certainty was assessed over up to 2.5 years of follow-up. The sample size per arm required to detect 30% attenuated rate in diagnostic certainty increase in trials was estimated over multiple time intervals. Of 254 patients with available longitudinal data, 61 patients had low diagnostic certainty at baseline (48 suggestive of PSP, 13 possible PSP) and multiple clinical visits (median: 3, range: 2-4). The cumulative percentage of patients increasing diagnostic certainty progressed with follow-up duration (30.4% at 6 months, 51.7% at 1 year, 80.4% at 2.5 years). The sample size required to detect 30% reduction in diagnostic certainty increase rate within 1 year was 163, slightly smaller than that required using the PSP rating scale. Most "suggestive of PSP" patients increased diagnostic certainty upon longitudinal follow-up, providing the first prospective multicenter validation of MDS-PSP diagnostic criteria. Our data support the design of trials tailored for these early-stage patients, suggesting the PSP rating scale and the diagnostic certainty increase rate as potential endpoint measures. © 2025 The Author(s). Movement Disorders published by Wiley Periodicals LLC on behalf of International Parkinson and Movement Disorder Society.

Longitudinal and Noninvasive Intracellular Recordings of Spontaneous Electrophysiological Activity in Rat Primary Neurons on Planar MEA Electrodes.

Presently, the in vitro recording of intracellular neuronal signals on microelectrode arrays (MEAs) requires complex 3D nanostructures or invasive and approaches such as electroporation. Here, it is shown that laser poration enables intracellular coupling on planar electrodes without damaging neurons or altering their spontaneous electrophysiological activity, allowing the process to be repeated multiple times on the same cells. This capability distinguishes laser-based neuron poration from more invasive methods like electroporation, which typically serve as endpoint measurement for cells. It is demonstrated that planar MEA electrodes, when combined with laser cell optoporation and live cell staining, can record spontaneous intracellular signaling from primary neurons in vitro. This approach allows for the detection of attenuated signals resembling positive monophasic intracellular action potentials. Recordings after laser optoporation also reveal subthreshold signals such as post-synaptic potentials that are essential for assessing neuronal network plasticity and connectivity. Moreover, the noninvasiveness of the process enables repeated intracellular recordings over multiple days from the same cells.

Resolving the Kinetics of Single-Walled Carbon Nanotube-Polyester Polyurethane Nanoparticle Conjugate Fluorescence Sensors toward Polymer Degrading Enzymes.

Single-walled carbon nanotubes (SWCNTs) are fluorescent materials that have been developed as sensors for measuring the activities of enzymes. However, most sensors to date rely on end-point measurement and empirical functions to correlate enzyme concentrations with fluorescence responses. Less emphasis is put on analyzing time-dependent fluorescence responses and their connections with enzymatic kinetics. Here, improved from our previous sensor design, we use trimethylchitosan-wrapped SWCNTs to measure the enzymatic degradation rate of Impranil nanoparticles, a polyester polyurethane model substrate. Through careful analysis of the characteristic time constant and saturation fluorescence, which are resolved from time-dependent brightening responses of the sensors, linear relations are found between fluorescence change rates and both enzyme concentrations and Impranil-to-SWCNT ratios, thus showing that the reaction is first-ordered toward both enzyme and substrate concentrations. The proposed sensor design and data analysis strategy can quantitively determine the relative enzymatic activity and provide insights into the kinetics of sensors and enzymatic reactions.

A Systematic Review of Multimodal Analgesic Effectiveness on Acute Postoperative Pain After Adult Cardiac Surgery.

To synthesise the best available empirical evidence about the effectiveness of multimodal analgesics on pain after adult cardiac surgery. A systematic review with meta-analysis. Indexed full-text papers or abstracts, in any language, of randomised controlled trials of adult patients undergoing cardiac surgery investigating multimodal postoperative analgesic regimen effect on mean level of patient-reported pain intensity at rest. Eight databases, via two platforms and three trial registries were searched from 1 January 1995 to 1 June 2024 returning 3823 citations. Of the 123 full-text papers assessed, 29 were eligible for inclusion. Data were independently extracted by a minimum of two reviewers in Covidence. There were 2195 participants, aged 60.4 ± 6.6 (range 40-79) years, who were primarily male (n = 1522, 76.1%), randomised in the included studies. Risk of bias was high and reporting quality was poor. Patient-reported pain was measured at rest in 28 (96.6%) trials. Data were suitable for pooled analysis from 10 (34.5%) of these trials with an average rest pain intensity of 3.3 (SD 1.5) in the control and 2.7 (SD 1.9) in the intervention groups, respectively. No trials compared combinations of nonopioid, opioid-agonist-antagonist, partial opioid agonists or full opioid agonists. Most trials (n = 11, 37.9%) compared two different full opioid options for less than 72 h (n = 24, 82.7%). Robust trials are needed to determine which multimodal analgesic combination will optimise patient recovery after adult cardiac surgery. There is an urgent need to test and refine high-quality end-point measures. Adequate assessment precedes ideal pain treatment. The findings from this review reveal neither are sufficient, and the impact of suboptimal pain management on postoperative recovery is grossly underinvestigated. The optimal combination of multimodal analgesics is unknown despite being recommended in best practice guidelines for enhanced recovery after cardiac surgery. Almost 30% of adults continue to experience ongoing pain up to a year after cardiac surgery, and findings from this review reveal a dearth of robust empirical evidence for optimal pain management, and heterogeneity in the way pain is assessed, measured and managed. This review provides a premise for robust trials focused on acute postoperative recovery in cardiac surgery and beyond. This review was conducted in accordance with the PRISMA-P statement. There was no patient or public contribution. PROSPERO: CRD42022355834.

Multi-stakeholder sessions on major innovation topics in rare disease clinical trials

BackgroundThe European Joint Programme on Rare Diseases aims to enhance the rare diseases research ecosystem by bringing together stakeholders such as research funders, institutions and patient organizations. Work Package 20 focuses on the validation, use and development of innovative methodologies for rare disease clinical trials. This paper reports on the outcomes of a retreat held in April 2023, where areas for innovation and educational needs in rare disease clinical trials were discussed in multi-stakeholder sessions.MethodsMulti-stakeholder sessions covered the topics: Future Educational System, Randomization in Rare Disease Clinical Trials, Endpoints in Rare Disease Clinical Trials and Using History Course Data. The sessions began with expert presentations to set the scene, followed by guided discussions facilitated by questions on a collaborative digital whiteboard. Participants wrote responses, which were then discussed live with the experts.ResultsTraining is needed for diverse stakeholders in rare disease clinical trials to enhance understanding and drive innovation. Challenges include a lack of standardized terminology for multiple endpoints, inadequate understanding of randomization in small sample studies and various obstacles in effectively using natural history data.ConclusionCreating a comprehensive and sustainable educational program for rare diseases clinical trial methodology requires strategic collaboration and adherence to FAIR principles. The workshop highlighted the need for innovations for topics in areas such as handling missing data, optimizing the extraction of information from small samples, remote endpoint measurement and new randomized inference techniques. Additionally, integrating innovations into tailored training programs is crucial for advancing the field.

Edridge Green Lecture 2022-demystifying clinical trials and regulatory approvals in drug development.

This article provides a comprehensive overview of clinical trial design and regulatory pathways essential for drug development, specifically in the context of retinal diseases. Key concepts include trial structure, efficacy and safety endpoints, and regulatory expectations from agencies like the FDA. It delves into recent regulatory advancements, such as the inclusion of low-luminance vision as a secondary endpoint and analyses case studies from age-related macular degeneration (AMD) trials. Approvals for key retinal drugs, such as ranibizumab and aflibercept, treatments for AMD and diabetic macular oedema, are discussed highlighting criteria like the 15-letter gain/loss in visual acuity as approvable/clinical meaningful efficacy endpoints. Insights into geographic atrophy (GA) and diabetic retinopathy trials showcase the evolving landscape, where anatomical endpoints and new drugs bring fresh challenges and opportunities. It also emphasizes the importance of academic-industry collaboration, citing instances of gene therapy development and innovative endpoint measures like the Multi-Luminance Mobility Test for retinal dystrophies. The overarching aim of this lecture was to demystify the process that spans the design of clinical trials to regulatory approval of drugs so that clinicians understand these complexities. In particular, it is important to understand the reasons behind selection of trial design, inclusion and exclusion criteria, primary and secondary efficacy endpoints and safety endpoints. Since this lecture, there have been important changes in this field including new guidance from the Food and Drug Administration (FDA) as well as lessons learnt from recent drug approvals that are included in this manuscript.

Creation of pH-Sensitive Bioreceptors for Affinity-Type Electrochemical Biosensor Applications

Introduction: Many disease states are characterized by the presence, absence, or change of molecular biological markers, or biomarkers. Common affinity-type recognition elements used in the detection of biomarkers are antibodies, either Immunoglobulins (IgGs) or recombinant fragments. While highly specific, the dissociation kinetics of antibodies highly favors the antibody-antigen complex, resulting in single, endpoint measurements [1]. Endpoint measurements are useful for ex vivo assays, but further antibody engineering is required for in vivo continuous monitoring. A key burden to overcome for using antibodies in continuous monitoring applications is the lack of binding site regeneration [2].Thus, we aim to create pH-modulable single chain fragment variables (scFvs), in which the KD of the scFv can be controlled to favor dissociation. We posit that through mutagenesis of residues within the complementarity-determining regions (CDRs) to histidine, we are able to modulate antibody binding kinetics through a change in pH [3,4]. There is a basis in the literature that supports this antibody design approach [5]. Those working with therapeutic antibodies currently employ similar techniques to effectively “recycle,” or regenerate the binding site of the antibody in vivo [6]. We seek to expand these techniques into the biosensing space to develop continuous, antibody-based biosensors. Methods: ClusPro was utilized to predict the antigen-binding residues in the CDRs of an anti-insulin scFv. From the predicted residues, we selected three amino acid residues to mutate to histidine with the intent of generating pH-modulable mutants. Three single mutants and a triple mutant containing all mutations were designed. The wild-type (WT) scFv, as well as the four mutants were all expressed as a soluble protein using Escherichia coli (E. coli) as a host microorganism. The binding ability of the WT and four mutants was assessed via two assays in differing pH with Bio-Layer Interferometry (BLI). The BLI assay workflow is as follows: biotinylated insulin was immobilized to Octet Streptavidin tips in buffers of either pH 7.4 or 6.0. Following immobilization, either the WT or mutants were introduced to bind to insulin. The resulting association and dissociation sensorgrams were recorded, and nonlinear fitting of the data was performed in GraphPad Prism 10.0. Results and Discussion: Compared to the WT, the triple histidine mutations in the CDR did not eliminate the scFv’s ability to bind to insulin. For the WT, the apparent KD decreased slightly between pH 7.4 and pH 6.0. This could be attributed to the change in pH affecting the streptavidin-biotin solution, or could be due to an artifact of the assay. Further studies would be needed to discern between the two hypotheses. However, there was a pronounced increase in the apparent KD of the triple mutant in a solution of pH 6.0 compared to pH 7.4. These results indicate the combination of mutations led to an observed pH-dependent binding. Further experimentation is needed with the single mutants to elucidate which residue is responsible for pH dependence, or if it is a combination of the three. Conclusions: We employed a rational mutation strategy to generate pH-dependent binding of an anti-insulin scFv. Upon a change in solution pH from 7.4 to 6.0, the triple mutant’s binding equilibrium is shifted, leading to an increase in KD. Further studies are needed to understand the reversibility of this interaction as well as to elucidate what specific residue interactions are contributing to this change.

Advancements in Cancer Research: 3D Models, Single‐Cell, and Live‐Cell Techniques for Better Insights

AbstractThe present review provides a comprehensive overview of the current state of in vitro cancer studies, focusing on recent advancements and ongoing in cell culture models and analyses techniques. Cancer cells grow in a complex and dynamic environment, interacting with various cellular components, such as stromal cells, cancer‐associated fibroblasts, immune cells, and the extracellular matrix (ECM). The ECM provides structural support and unique characteristics essential for tumorigenesis. Accurately modeling this intricate tumor microenvironment and precisely analyzing cell–cell and cell–ECM interactions are crucial for understanding cancer progression and therapeutic responses. Consequently, oncology research is advancing toward a) three‐dimensional tumor models, b) single‐cell level analyses, and c) live‐cell analyses. This review aims to elucidate current knowledge in this field, emphasizing the benefits these innovative approaches offer over traditional two‐dimensional models, bulk analyses, and endpoint measurements.

A Three-Month Clinical Trial on the Efficacy of Hyaluronic Acid Adjunctive Non-Surgical Therapy for Periodontitis in Patients with Type 2 Diabetes Mellitus.

Background/Objectives: Conventional periodontal treatment for patients with diabetes has shown promising results, primarily focusing on glycated hemoglobin (HbA1c) levels as an endpoint measure. The properties of hyaluronic acid (HA) have been harnessed in various periodontal therapies, and it is a promising agent also in a non-surgical approach. The aim of this clinical trial was to assess the efficacy of hyaluronic acid in a local adjunctive non-surgical treatment for periodontitis in patients with type 2 diabetes. Methods: Eighty adult participants with well-controlled type 2 diabetes (HbA1c 7% (53 mmol/mol) or less) took part in the trial. The clinical parameters of periodontitis as well as the glycated hemoglobin (HbA1c) levels were evaluated, and an analysis of the potential differences between the control (placebo) and intervention (HA) groups was performed. Results/Conclusions: A decrease in all the clinical values of periodontitis after treatment was observed in the vast majority of patients in both groups. Differences in the clinical parameters were observed 12 weeks after the intervention between the patients in the placebo and HA therapy groups. Bleeding on probing (BoP) was reduced in the control group to 15-25% and was approximately 5.5% more in the intervention group (9.5-18.25%). The clinical attachment level (CAL) decreased 1 mm more in the HA therapy group (1-2 mm) than in the no adjunctive treatment group (2-3 mm). The probing depth (PD) was reduced similarly in both groups (3-3.75 mm). Due to the bilateral relationship between diabetes and periodontitis, healthcare professionals seek advancements in managing periodontal inflammation. The results of this study indicate that non-surgical periodontal treatment with HA as an adjunctive agent is worth considering in the therapy for patients with diabetes.

A novel method for real-time inhalation toxicity assessment in mice using respirometric system: A promising tool for respiratory toxicology

Inhalation toxicity assessment is a crucial tool for the identification and classification of hazardous materials like volatile organic carbons, aerosols, and particulate matter. Unlike traditional acute inhalation toxicity studies that use mortality as an endpoint, the Fixed Concentration Procedure (FCP) emphasizes "evident toxicity" by monitoring behavior, weight, and food intake. This reduces reliance on mortality but doesn't directly address respiratory system impact. The present study introduced a respirometer-based inhalation toxicity and respiratory status assessment method. The toxicity evaluation system integrated a respirometric system with an animal exposure chamber, enabling real-time monitoring of oxygen consumption. The ICR mice were exposed to various concentrations of benzene (10, 20, 40, and 80 mg/L of air), toluene (7.5, 15, 30, and 60 mg/L of air), and xylene (7.5, 15, 30, and 60 mg/L of air). The respiration rate decreased by 70 % and 69 % for benzene (80 mg/L of air) and toluene (60 mg/L of air), respectively, with EC50 values of 32.5 mg/l and 21.2 mg/L based on oxygen consumption. Xylene did not exhibit EC50 values at the tested concentrations. However, the oxygen consumption rate significantly decreased (46 %) at high concentrations (60 mg/L of air), indicating sub-lethal toxicological effects. Furthermore, the present study was also validated in the bleomycin-induced idiopathic pulmonary fibrosis (IPF) model, demonstrating its reliability as a respiratory impairment marker. The results exhibited a strong correlation between weight loss and less oxygen consumption in the BLM group (bleomycin-induced) as compared to the SHAM group (control), which was confirmed by histological examination and protein marker analysis. The results suggest the potential use of oxygen consumption as an endpoint measurement in inhalation toxicity assessment tests without animal sacrifice, and the present study could be useful for providing valuable insights into disease progression and pharmacological interventions.

Consensus definition of a radiologically healed fistula on magnetic resonance imaging in perianal Crohn’s disease: an international Delphi study

IntroductionPerianal fistulising Crohn’s disease (pfCD) is a distinct and debilitating phenotype seen in around one-third of patients with CD. Clinical trials in pfCD are increasingly using magnetic resonance imaging (MRI)...

The effect of vitamin D3 supplementation on vitamin D status and associated health outcomes in children

Sufficient vitamin D status is required in childhood for normal skeletal health and the development of muscle and motor function(1). Vitamin D primarily acts through the promotion of calcium absorption from the gut and mobilisation from bone tissue(2). Childrens’ vitamin D dietary intakes are currently below the recommended guidelines within the UK and Ireland suggesting a need for vitamin D3 supplementation to prevent the risk of developing vitamin D deficiency(3). The aim of this study was to examine the effect of 12 weeks’ supplementation with 10µg/d vitamin D3 vs placebo control on vitamin D status and to determine if improved vitamin D status impacted muscle function and cognition in children aged 4-11 years.In the D-VinCHI randomised, double-blind, placebo-controlled trial, healthy children (n = 118; mean age 8.1 ± 1.8 y; 51% girls) were randomly assigned to either placebo or 10µg/day of vitamin D3 for 12 weeks (year-round). Baseline and endpoint measures included anthropometric measures, hand grip strength, balance, and cognitive assessment. Blood samples were analysed for plasma 25hydroxyvitamin D [25(OH)D], and parathyroid hormone. Vitamin D consumption from food sources was assessed via a 13-item food frequency questionnaire.Following the 12-week intervention vitamin D status [25(OH)D concentration] increased in the treatment group from 66.31 ± 17.25 nmol/L to 69.04 ± 16.92 nmol/L. Change in status was significantly different compared to the placebo group within which a decrease in 25(OH)D was observed from 63.67 ± 19.48 to 56.29 ± 18.58 nmol/L, p<0.001. Supplementation with vitamin D3 prevented deficiency of plasma 25(OH)D during the winter months. Vitamin D3supplementation had a positive effect on cognitive function, improving simple movement time from 707 ± 380 to 599 ± 207 milliseconds, compared to the placebo group within which time increased from 680 ± 284 to 728 ± 372 milliseconds, p = 0.017. Vitamin D mean dietary intake from food sources alone was 2.68µg/day. There was no effect of supplementation on muscle function.Vitamin D3 supplementation maintained year-round sufficiency in children and importantly, prevented deficiency during the extended winter months. Vitamin D supplementation may enhance cognitive function via improvements in attention and psychomotor speed. The reported intakes of vitamin D food sources were low and were well below the current dietary reference value for this age group. Further health promotion policies and (bio)fortification strategies should be considered within this age group to maximise vitamin D status for optimal growth and development.

Development of an Improved Thiosulfate-Utilizing Denitrifying Bacteria-Based Ecotoxicity Test with High Detection Sensitivity and Reproducibility.

Microorganism-based ecotoxicity assessment has been widely used as a reliable tool showing direct biochemical impacts of contaminants on ecosystems and the environment. The present study aimed at developing a thiosulfate-utilizing denitrifying bacteria (TUDB)-based ecotoxicity test with high detection sensitivity and favorable reproducibility. To achieve this goal, existing TUDB toxicity tests were improved by employing a pure culture of Thiobacillus thioparus ATCC 8158 and optimizing test conditions, particularly in terms of inoculated microbial biomass, incubating temperature, and operational pH. From control tests, it was found that 4 h is a sufficient processing time for TUDB test kits. As a result of optimization, 20 mg VSS/L of initial bacterial biomass, 25 °C of incubating temperature, and 6 of operational pH were determined as the most favorable test conditions, providing enhanced detection sensitivity and reproducibility. Under these optimal test conditions, I conducted toxicity tests for diverse toxic metals and obtained 0.65 ± 0.03, 1.09 ± 0.04, 1.21 ± 0.07, 0.13 ± 0.01, 0.56 ± 0.04, 1.42 ± 0.03, 0.98 ± 0.02, and 2.12 ± 0.05 mg/L of 4 h EC50 values for Ag+, As3+, Cd2+, Cr6+, Cu2+, Hg2+, Ni2+, and Pb2+, respectively. These EC50 values are substantially lower than those from earlier TUDB tests, demonstrating the high detection sensitivity of the current TUDB tests. Moreover, the present TUDB tests attained very low coefficient of variation (CV) values (1.6-6.3%) for the EC50, showing favorable reproducibility of the test methodology. In addition, the current TUDB toxicity tests offer numerous advantages for ecotoxicity assessment, including versatility for diverse test samples, no requirement for advanced equipment, and no distortion of end-point measurement. These refinements render the TUDB tests a favorable ecotoxicity assessment with enhanced sensitivity and reproducibility.

ToxProfiler: A novel human-based reporter assay for in vitro chemical safety assessment

In vitro chemical safety assessment often relies on simple and general cytotoxicity endpoint measurements and fails to adequately predict human toxicity. To improve the in vitro chemical safety assessment, it is important to understand the underlying mechanisms of toxicity. Here we introduce ToxProfiler, a novel human-based reporter assay that quantifies the chemical-induced stress responses at a single-cell level and reveals the toxicological mode-of-action (MoA) of novel drugs and chemicals. The assay accurately measures the activation of seven major cellular stress response pathways (oxidative stress, cell cycle stress, endoplasmic reticulum stress, ion stress, protein stress, autophagy and inflammation) that play a role in the adaptive responses prior to cellular toxicity. To assess the applicability of the assay in predicting the toxicity MoA of chemicals, we tested a set of 100 chemicals with well-known in vitro and in vivo toxicological profiles. Concentration response modeling and point-of-departure estimation for each reporter protein allowed for chemical potency ranking and revealed the primary toxicological MoA of chemicals. Furthermore, the assay could effectively group chemicals based on their shared toxicity signatures and link them to specific toxicological targets, e.g. mitochondrial toxicity and genotoxicity, and different human pathologies, including liver toxicity and cardiotoxicity. Overall, ToxProfiler is a quantitative in vitro reporter assay that can accurately provide insight into the toxicological MoA of compounds, thereby assisting in the future mechanism-based safety assessment of chemicals.

Safety and efficacy of canine gonadal tissue-derived mesenchymal stem cells for early myxomatous mitral valve disease.

This study explored the potential efficacy and safety of therapy with mesenchymal stem cells (MSC) derived from gonadal tissue to address the early stage of myxomatous mitral valve disease (MMVD), the predominant cardiac condition in dogs. Sixteen dogs diagnosed with MMVD B1 were enrolled in this trial and assigned to either a control group (control group, n = 10) or a group that received MSC derived from gonadal tissue (treatment group, n = 6). In the treatment group, allogeneic MSC derived from gonadal tissue (1 × 106 cells/kg) were intravenously administered at monthly intervals for five or more sessions. Data were compared at baseline and at the endpoint 1-year intervals. The efficacy was assessed using echocardiography, thoracic radiography, NT-proBNP, and the duration from B1 diagnosis to B2 transition to evaluate its effect on MMVD stage progression. Safety was evaluated through physical examinations, blood tests, imaging studies, and monitoring of adverse events. After 1 year of observation, the control group exhibited deteriorating echocardiographic parameters, whereas the treatment group displayed no substantial differences between baseline and endpoint measurements. Notably, a statistically significant disparity was noted in the left atrial diameter (p < 0.05) and E-wave velocity (p < 0.05) between the two groups, indicating a favorable impact of MSC derived from the gonadal tissue on left atrial pressure. Additionally, in contrast to the control group, the treatment group demonstrated delayed progression to MMVD stage B2, enabling them to prolong their disease duration without requiring cardiac medication (p = 0.038). In quality of life (QoL) metrics following MSC treatment, appetite showed a statistically significant improvement, increasing from 4 to 4.83 (p < 0.05). Treatment with gonadal tissue-derived MSCs significantly delayed MMVD stage progression, highlighting the broad potential of MSC derived from gonadal tissue for treating complex veterinary conditions.

A-085 Comparing CO2 stability in patient samples and proficiency testing material

Abstract Background Total carbon dioxide (tCO2) measurements are important in the assessment of acid-base disorders. The collection and storage conditions of specimens are important preanalytical factors that can influence the accuracy of tCO2 measurements. Proficiency testing (PT) materials are used to assess the accuracy and reliability of a laboratory’s test performance. Unsuccessful performance on PT challenges can be an indicator of procedural issues. However, the stability of tCO2 in PT material compared to physiologic specimens is unknown and no guidance is provided to laboratories on how to handle PT samples for tCO2 measurement. This study aims to compare tCO2 stability in PT material with that of plasma samples when exposed to ambient air at room temperature. Methods Using enzymatic endpoint measurement on Vitros 4600 Chemistry System, tCO2 was measured in PT material (n = 5) and patient samples (n = 6) at baseline and after exposure to air for 15, 30, 60, 90, and 120 minutes. 500μL of each sample was aliquoted into EZ-nest tube adapters and baseline measurements were immediately analyzed. Average percent difference from baseline was calculated and &amp;gt;-10% bias was considered acceptable. Results The average percent difference from baseline in PT samples exceeded -10% as early as 30 minutes; whereas -3% bias was observed in patient samples at 60 minutes. Results are summarized in the table below. Conclusions The assessment of tCO2 stability ensures the reliability of laboratory measurements. Our results indicate tCO2 in PT material is not stable when exposed to air beyond 15 minutes, however tCO2 in most patient samples remained stable beyond 120 minutes. Understanding the stability limits of tCO2 in PT material allows us to devise workflows that prevent sample degradation during PT events. By establishing best practices for maintaining tCO2 integrity in PT material, laboratories can enhance the accuracy of proficiency testing results.

Responsiveness of different disease activity indices in moderate-to-severe ulcerative colitis.

Clinical, endoscopic, histological, and composite instruments are currently used to measure disease activity in patients with ulcerative colitis (UC). We compared the responsiveness of the Mayo Clinic score (MCS), modified MCS (mMS; excluding physician global assessment), partial MCS (pMS; MCS without endoscopic subscore), Robart's Histopathology Index (RHI), and UC-100 score to change after ustekinumab treatment in patients with moderately to severely active UC. Post hoc analysis of the phase 3UNIFI induction trial (ClinicalTrials.gov: NCT02407236) was conducted. Participants with moderately to severely active UC were randomized to receive ustekinumab or placebo. Treatment assignment was the criterion to assess responsiveness, which was quantified using the probability of a treated participant having a larger change in score than a placebo participant, termed the win probability (WinP), and estimated using nonparametric methods. The UC-100 score demonstrated large responsiveness (WinP 0.72 [95% confidence interval: 0.66-0.78]), and the MCS (0.68 [0.62-0.73]), mMS (0.69 [0.63-0.75]), and pMS (0.65 [0.59-0.71]) demonstrated similar effect sizes. Of the component items of the Mayo score, the endoscopic subscore (WinP 0.76 [0.69-0.82]) and the stool frequency subscore (WinP 0.74 [0.69-0.79]) were the most responsive. The Inflammatory Bowel Disease Questionnaire (IBDQ) quality-of-life questionnaire was also responsive (WinP 0.78 [0.72-0.82]). UC disease activity indices are similarly responsive. Depending on the treatment setting, time point of evaluation, and feasibility of measurement, different scores may be used to demonstrate response. These results support the use of mMS as a composite primary endpoint, incorporating both patient-reported and endoscopic outcome measures. The UC-100 score may be more appropriate in settings that also routinely incorporate histological evaluation. There is no funding for this study.

Semaglutide treatment for PRevention Of Toxicity in high-dosE Chemotherapy with autologous haematopoietic stem-cell Transplantation (PROTECT): study protocol for a randomised, double-blind, placebo-controlled, investigator-initiated study

IntroductionCancer treatment with high-dose chemotherapy damages the mucosal barrier of the gastrointestinal (GI) tract and is associated with severe toxicity involving mucositis, severe inflammation and organ dysfunction. Currently, there is...

Comparison of ustekinumab, infliximab and combination therapy in moderately to severely active ulcerative colitis - a study protocol of a randomized, multicenter, head-to-head COMBO-UC trial.

Ulcerative colitis (UC) is a chronic inflammatory bowel disease with a complex etiology that affects the large intestine. Characterized by chronic, bloody diarrhea, UC can lead to severe complications, including an increased risk of colorectal cancer. Despite advancements in conservative treatment, including biologics like anti-TNF agents and ustekinumab (UST), many patients do not achieve full remission. Dual targeted therapy (DTT) combining infliximab (IFX) and UST is a promising approach to improve treatment outcomes. This prospective, randomized, multicenter, head-to-head controlled trial will evaluate the efficacy and safety of UST, IFX, and combination therapy (UST + IFX) in 172 patients with moderate to severe active UC across eight gastroenterology centers in Poland. The study includes a 14-16 week remission induction period followed by a 52-week maintenance phase. Patients will be randomly assigned to one of three treatment arms: IFX monotherapy, UST monotherapy, or IFX + UST combination therapy. Primary endpoint is clinical and endoscopic remission post-induction. Secondary endpoints include clinical response, biochemical remission, histological remission, and quality of life assessments using the Inflammatory Bowel Diseases Questionnaire and 36-Item Short Form Survey. Safety will be monitored through adverse event and serious adverse event reporting. This trial aims to determine whether combining IFX and UST can achieve higher remission rates and better long-term outcomes compared to monotherapy. The results could provide crucial insights into the optimal use of biologic agents in UC treatment, potentially establishing DTT as a standard therapy. The study's design, including extensive follow-up and robust endpoint measures, will contribute to understanding the therapeutic potential and safety profile of this combination therapy.

Development of a new method for analyzing the behavior of rapidly dissolving films in contact with liquids

Oral Thin Film (OTF) is a promising dosage form, easy to use and patient-friendly drug delivery systems suitable for every consumer profile. This dosage form ensures safety, stability, and acceptability of the drug, that's why it can be a good alternative for tablets and capsules for children and elderly therapies. As no detailed monography of OTFs including characterization methods and uniform requirements has been presented in any of the pharmacopoeias to date, this study focuses on the development of an innovative analyzing method to characterize the OTF behavior and particularly to determine its disintegration time by goniometry. These parameters are key for the development of OTFs and for studying, at an early stage, the impact of formulation and process variables on product quality and impacting Critical Quality Attributes (CQA) by a Quality by Design approach.Different model OTF preparations were investigated as models to determine the suitability of the disintegration test systems. These placebos OTF were used to compare the disintegration times obtained by the tablets pharmacopeial method (conducted in 800 mL of distilled water) and the novel proposed method. As expected, the disintegration times determined by the proposed method were similar than those obtained by the conventional disintegration test. Moreover, they seem more reliable to the in vivo behavior because of the small amount of solvent used. The experiments showed acceptable reproducibility (low variation within sample replicates due to a well-defined determination of the measurement endpoint). This method provides clear endpoint detection and is applicable for different types of OTFs based on different polymers. Moreover, a better understanding of the physical-chemical properties of oral thin films towards water could be obtained. This new method of analysis could provide a valuable approach to the development of new OTF formulas, thanks to the different information that can be obtained, such as critical energy, free energy and polar and dispersive components.