Bioanalytica (Switzerland)

This study aims to examine the implementation of Perplexity in the preparation of Assessment as Learning as an innovation in technology-based learning assessment. The application of Perplexity allows teachers and students to optimize the formative assessment process through the use of accurate, relevant, and easily accessible information sources. The purpose of this study is to describe the steps of implementing Perplexity and the preparation of Assessment as Learning in learning at Madrasah Tsanawiyah Negeri 1 Sukoharjo, Madrasah Tsanawiyah Negeri 2 Sragen, and Madrasah Tsanawiyah Negeri 1 Klaten. This research method uses a qualitative approach. The subjects of the study were teachers at Madrasah Tsanawiyah Negeri 1 Sukoharjo, Madrasah Tsanawiyah Negeri 2 Sragen, and Madrasah Tsanawiyah Negeri 1 Klaten. The informants of this study were teachers. Data collection techniques were interviews, observation, and documentation. The results of this study are: 1) The application of Perplexity in the preparation of Assessment as Learning in learning at Madrasah Tsanawiyah Negeri 1 Sukoharjo, Madrasah Tsanawiyah Negeri 2 Sragen, and Madrasah Tsanawiyah Negeri 1 Klaten is carried out through the following stages, namely: a) Visit the official Perplexity website at www.perplexity.ai; b) Select the desired interaction mode. This mode adjusts the type of question and analysis needs; c) Type the question directly in the chat column. Then ask for the preparation of Assessment as Learning; d) Perplexity will provide answers along with the main reference sources from the search results; e) Search results and discussions can be saved in a special collection, complete with names, descriptions, and advanced instruction settings for subsequent use; 2) Steps for preparing Assessment as Learning by utilizing Perplexity: 1) Integration with a learning management system (LMS); 2) Development of adaptive tests; 3) Providing instant feedback; 4) Learning data analysis; 5) Personalization of learning; 6) Teacher and student training; 7) Periodic supervision and evaluation. The conclusion of this study is that the use of Perplexity in the development of AfL can improve students' learning independence, strengthen reflective and metacognitive skills, and assist teachers in designing assessment instruments based on data and conceptual understanding. Thus, the application of Perplexity not only supports assessment effectiveness but also encourages the creation of adaptive, collaborative, and student-centered learning.

Plasma kallikrein (pKal) is a trypsin-like serine protease involved in the kallikrein-kinin, renin-angiotensin, and complement pathways, making it an attractive target for diseases, such as hereditary angioedema, diabetic mellitus complications, and cerebrovascular disorders. As part of an internal program to develop orally bioavailable small-molecule pKal inhibitors, we report lead optimization efforts within the spirocarbamate scaffold, highlighting a structure-based drug design strategy to engineer hydrogen bond interactions with N-benzyl aminopyrazoles. Additionally, mitigation of time-dependent inhibition (TDI) liability and optimization of the overall profile were achieved through a two-pronged strategy: (1) incorporating increased Fsp3 modifications via N-alkylation and (2) leveraging torsional strain in N-aryl analogs.

The high variability in efficacy and safety of antichagasic chemotherapy involving benznidazole (BNZ) may be due to pharmacokinetic-related factors. The study evaluated the impact of experimental acute and chronic infections by the Berenice-78 strain of Trypanosoma cruzi on the pharmacokinetics of BNZ in dogs. Twenty-seven mongrel dogs were divided into the following groups: acute infection state treated with BNZ, chronic infection state treated with BNZ, acute and chronic positive controls (not treated with BNZ), and a healthy group treated with BNZ. They were evaluated at (1) basal state, (2) during infection without treatment, for cytokines panel (IL-6, interferon γ (IFN-γ), IL-10 and tumor necrosis factor α (TNF-α)) evaluation, and (3) during BNZ steady-state levels at 10, 30, 40, and 60 days after start of treatment with oral BNZ 3.5 mg/kg b.i.d. administration, in acute and chronic T. cruzi-infection, for BNZ pharmacokinetics and cytokines panel evaluation, and (4) 30 days after BNZ treatment end for cytokines evaluation. BNZ levels in serum samples were quantified using a curve range of 0.1-100 μg/mL in plasma by high-performance liquid chromatography with diode array detection (HPLC-DAD) analysis. Pharmacokinetic parameters remained stable during treatment phases, indicating no autoinduction or inhibition. Acute infection pharmacokinetics resembled that of healthy controls. Chronic infection significantly increased C max, C ss, and AUC0-12, while decreasing Vd/F and CL/F compared to both healthy and acute groups. Notably, IL-6 levels were elevated ∼7-fold during chronic infection. These data suggest that chronic inflammatory status modulates BNZ disposition, likely via IL-6 mediated P-glycoprotein inhibition. Understanding these pharmacokinetic changes is critical for optimizing BNZ dosing strategies in Chagas disease management.

ABSTRACTBexicaserin is an investigational molecule that is a highly selective superagonist of the 5‐hydroxytryptamine 2C (5HT2c) serotonin receptor subtype and in Phase 3 clinical studies for the treatment of seizures associated with developmental and epileptic encephalopathies (DEEs). We have validated a simple LC–MS/MS method for the quantitative estimation of bexicaserin in human cerebrospinal fluid (CSF). The chromatographic separation of bexicaserin and the IS was achieved on an Acquity HSS T3, C18 column using a 5.5‐min gradient program. The sample preparation involves the dilution extraction of bexicaserin and the isotope labeled internal standard (13CD2‐bexicaserin; IS) from 25 μL of human CSF. The calibration curve was linear from a range of 0.1–100 ng/mL. The method was fully validated as per the regulatory guidelines. The intraday and interday precision for bexicaserin was in the range of 6.3%–8.7% and 6.0%–8.8%. Bexicaserin intraday and interday bias was in the range of −15.1% (LLOQ) to 12.0% and −10.0% to 1.2%. Stability studies showed that bexicaserin was stable on bench for 24 h, in autosampler for over 50 h, and extract stability at 6°C for 76 h. Bexicaserin was stable after five freeze–thaw cycles and long‐term storage at −20°C for 108 days and −80°C for 102 days.

Rationale:Tuberculosis vaccines aim to elicit protective immunity, with IFNγ-producing CD4⁺ T cells considered central mediators of host defense. In murine models, IFNγ activates macrophages to induce inducible nitric oxide synthase (iNOS), resulting in nitric oxide–dependent control of Mycobacterium tuberculosis (Mtb).Objective:To define the relevance of IFNγ-induced iNOS activity in humans and other natural host of virulent mycobacterial species.Methods:We systematically compared IFNγ-induced effector responses in Mtb-infected myeloid cells across species. Using bulk RNA sequencing, functional infection assays, and nitric oxide measurements, we assessed IFNγ responsiveness in human and mouse macrophages. These analyses were extended to monocytes from seven mammalian species and complemented by reanalysis of publicly available single-cell RNA-sequencing datasets and spatial proteomic imaging of tuberculous granulomas.Measurements and Main Results:Single-cell transcriptomic reanalysis revealed minimal NOS2 expression in myeloid cells from human and non-human primate granulomas. In vitro, IFNγ pretreatment failed to induce NOS2 transcription, iNOS activity, or Mtb growth restriction in human macrophages, in stark contrast to murine cells. Across species, iNOS activity was largely restricted to mice, with limited induction in cattle monocytes. Instead, human respiratory epithelial cells consistently expressed NOS2, and multiplexed ion beam imaging localized iNOS protein to epithelial compartments adjacent to granulomatous lesions.Conclusion:IFNγ signaling is uncoupled from iNOS induction in primate myeloid cells and epithelial compartments represent dominant sources of iNOS in human tuberculosis. These findings, challenge murine macrophage–centric paradigms and IFNγ-based correlates used in TB vaccine development and central to pathophysiology of tuberculosis and other pneumonias.

High stroma content, as measured by tumor-stroma ratio (TSR), is generally a negative prognostic parameter for epithelial cancers, including sporadic colorectal cancer (sCRC). Inflammatory bowel disease patients have higher risk for colorectal cancer than the background population. Evidence suggests that this colitis-associated colorectal cancer (CAC) is more aggressive and occurs at younger age than sCRC. CAC also differs from sCRC in oncogenesis and prognosis. This study tests the hypothesis that TSR in CAC tumors correlates with survival. Age at CAC diagnosis relative to TSR was also explored. TSR was quantified in 36 CAC cases. In routine hematoxylin–eosin staining, the amount of stroma was estimated in categorical steps of 10% increments per image field. The area with highest amount of stroma and tumor tissue at all quadrants of the visual field boundary was scored for TSR. For statistical analysis, tumors were divided into stroma-high (> 50%) or stroma-low (≤ 50%). Of all cases, 22 were stroma-high and 14 were stroma-low. Five-year survival in the stroma-high group was 32% (n = 22), compared to 71% (n = 14) in the stroma-low group (p = 0.049). High stroma content was more frequent if cancer diagnosis was before 60 years of age (17/23) compared to after 60 years of age (5/13). Despite differences in oncogenesis and tumor biology in CAC compared to sCRC, high stroma content also predicts worse outcome in CAC and is particularly common in younger patients.

IntroductionSputum biomarker measurements are used to measure airway inflammation in COPD patients. We have previously validated a Luminex assay able to quantify 15 analytes in COPD sputum supernatant. This assay demonstrated sputum protein expression profiles associated with neutrophilia, airway bacterial colonisation and current smoking in COPD.MethodsWe report repeat sputum supernatant analysis at 6 months from a sub-group of COPD patients who participated in the original study. 48 COPD patients provided a repeat sputum sample at 6 months. 15 panel analytes were detectable.ResultsRepeated sputum cytokine measurements showed a significant positive correlation between baseline and 6 months for all analytes except IL-1RA with intraclass correlation coefficient (ICC) indicating good to excellent repeatability. IL-1β, IL-2, IL-8, IL-17A, G-CSF, MIP-1α, MIP-1β and TNF-α were significantly correlated with sputum neutrophil percentage at 6 months. IL-1β, IL-4, IL-8 and G-CSF were significantly increased in Haemophilus influenzae colonised patients and IL-1β, IL-4, IL-8, G-CSF, IFN-γ, IP-10, MCP-1, MIP-1α, MIP-1β and TNF-α were significantly higher in ex-smoking COPD patients.DiscussionA multiplex immunoassay used at repeated visits in COPD patients showed a high degree of reproducibility for the majority of analytes. There were reproducible inflammatory signatures in sputum associated with clinical characteristics in COPD patients.

The implementation of variable pathlength spectroscopy in ultrafiltration and diafiltration processes addresses critical limitations in nucleic acid manufacturing through real-time concentration monitoring and automated process control. This article presents real-world case studies demonstrating significant time savings and achieving less than 5% deviation from target concentration endpoints through integrated tangential flow filtration systems.

Abstract. The Tren Maya project, a new railway connecting major population centres and tourist sites around Mexico’s Yucatan peninsula, impacted vast tracts of undeveloped land, ripe with archaeologically significant sites and landscapes. This effort necessitated the mobilization of Mexico’s archaeological authorities for the evaluation of large areas with tight timelines, as well as the creation of new museums along this railway network, consolidating and cataloguing previously scattered artifact collections. Authorities invested heavily in 3D survey products to meet the rigorous requirements of the project, resulting in the creation of many 3D aerial and terrestrial LiDAR, photogrammetry, structured light datasets. These survey data offer critical insight into their subjects and have wide potential for re-use by external parties, and are of great interest to the public at large, but, their inherent complexity and density create significant barriers for accessibility. We present a scalable and future-facing web-based system and archival framework enabling the long-term accessibility, visualization, and interoperability of critical 3D data assets with existing research and site management pipelines, game/xr/metaverse environments, and machine learning training applications.