Journals
2025 EN
ŚliwaDominiak Joanna · Czechowska Kamila · Blanco Alfonso
+5 more
ABSTRACT Flow cytometry (FC) is a versatile and powerful tool in microbiology, enabling precise analysis of single cells for a variety of applications, including the detection and quantification of bacteria, viruses, fungi, as well as algae, phytoplankton, and parasites. Its utility in assessing cell viability, metabolic activity, immune responses, and pathogen‐host interactions makes it indispensable in both research and diagnostics. The analysis of microbiota (community of microorganisms) and microbiome (collective genomes of the microorganisms) has become essential for understanding the intricate role of microbial communities in health, disease, and physiological functions. FC offers a promising complement, providing rapid, cost‐effective, and dynamic profiling of microbial communities, with the added ability to isolate and sort bacterial populations for further analysis. In the probiotic industry, FC facilitates fast, affordable, and versatile analyses, helping assess both probiotics and postbiotics. It also supports the study of bacterial viability under stress conditions, including gastric acid and bile, improving insight into probiotic survival and adhesion to the intestinal mucosa. Additionally, the integration of Machine Learning in microbiology research has transformative potential, improving data analysis and supporting advances in personalized medicine and probiotic formulations. Despite the need for further standardization, FC continues to evolve as a key tool in modern microbiology and clinical diagnostics.
Journals
2025 EN
Yue Alice · Brinkman Ryan R. · Nash Veronica
+9 more
Abstract Flow cytometry is a powerful tool for analyzing diverse cellular properties, making it essential in immunology research, clinical trials, and diagnostics. Integrating artificial intelligence (AI) into flow cytometry has the potential to enhance various aspects of assay development and application, including reagent selection, instrument standardization, panel and assay design, data analysis, quality controls, and knowledge dissemination. This paper provides a review of current AI applications in flow cytometry and explores the potential future directions for AI integration in the field.
Journals
2025 EN
Amparo Tatiane Roquete · Anunciação Kamila de Fátima da · Almeida Tamires Cunha
+2 more
ABSTRACT α‐Lapachone (aLAP) and β‐lapachone (bLAP) are noteworthy anticancer naphthoquinones. The chemoresistance observed in bladder cancer represents a global health concern, with relation to mutations in the TP53 gene and alterations in the expression of long noncoding RNA (lncRNAs). This study evaluated the effects of aLAP and bLAP on bladder tumor cell lines with different TP53 statuses: RT4 low‐grade tumor with wild‐type TP53 ), T24 and J82 (high‐grade tumor with mutation in the TP53 gene). Cytotoxicity was assessed using the MTT reduction method and cell migration by scratch assay, while clonogenic survival and cell cycle were evaluated through cell colony counting and flow cytometry, respectively. The expression of lncRNAs linked to bladder cancer and associated with tumor progression and prognosis ( JHDM1D‐AS1 , SBF2‐AS1 , CDT‐2132N18.2 , and RP11‐363E7.4 ) and the JHDM1D gene was evaluated through RT‐qPCR. bLAP demonstrated greater cytotoxicity than aLAP. Its inhibitory effects on clonogenic survival, migration, and the cell cycle were observed in all cell lines and were related to the modulation of lncRNAs expression. A reduction in lncRNA SBF2‐AS1 and JHDM1D gene expression was observed in RT4 cells, accompanied by an increase in lncRNA RP11‐363E7.4 . Conversely, in the cells with mutated TP53 (J82), a reduction in JHDM1D‐AS1 and JHDM1D was observed. The downregulation of JHDM1D‐AS1 and SBF2‐AS1 , along with the upregulation of RP11‐363E7.4 , may be associated with the observed inhibition of proliferation and cell migration following bLAP treatment. The antiproliferative effects of bLAP in bladder cancer cells are independent of TP53 statuses, yet occur through a distinct action mechanism, with variations in lncRNAs expression.
Journals
2025 EN
Hartopo Anggoro Budi · Achyar Arinal Chairul · Bagaswoto Hendry Purnasidha
+17 more
Abstract Aims This study aimed to conduct a phase 2 proof‐of‐concept and safety study to evaluate the effect of ENIBARCIMAB (EN), a non‐neutralizing humanized monoclonal antibody targeting the N‐terminus of adrenomedullin (ADM), administered immediately after stabilization with standard of care (SoC) treatment, in patients hospitalized for acute heart failure (AHF). Methods and results This prospective, open‐label, controlled, interventional, multicenter, dose‐escalation study was conducted at two cardiology sites in Indonesia. Patients were divided into two interventional groups sequentially receiving 0.5 mg/kg (SoC + EN 0.5 mg/kg, n = 10; first cohort) and 2 mg/kg (SoC + EN 2 mg/kg, n = 10; second cohort) of EN via 1‐h intravenous (IV) infusion within 48 h after admission for AHF. The control group ( n = 10) was treated with SoC therapy for AHF therapy. All patients were monitored continuously within 24 h post‐infusion and subsequent daily until discharge. Treatment‐related serious adverse events (SAEs) were recorded during hospitalization and up to 90 days after discharge. Both EN dosages were well‐tolerated, and no significant safety issues were identified during hospitalization and up to 90 days of follow up. SAEs occurred in 10% of patients in each EN group but were deemed not related to treatment. No significant differences in the occurrence of SAEs were found between the groups. Five deaths occurred: three (30%) in the control group as compared with two deaths (20%) in the SoC + EN 2 mg/kg group. EN led to a significant increase in plasma bio‐ADM levels within 24 h post‐infusion, with the SoC + 2 mg/kg group showing the highest increase. Within 1 h from IV EN infusion, SoC + EN 2 mg/kg compared with 0.5 mg/kg, resulted in a significant percentage reduction in systolic, diastolic blood pressure, and mean arterial pressure. However, it did not result in severe hypotension and the need for drug discontinuation. Conclusions In this pilot safety study of patients hospitalized for AHF, IV infusion of EN 0.5 and 2 mg/kg increased circulating plasma bio‐ADM levels and was well‐tolerated without treatment‐related SAEs occurring during hospitalization and up to 90 days after discharge.
Journals
2025 EN
Koszelska Kamila · Wypych Monika · Kciuk Mateusz
+3 more
In the current study, an evaluation was conducted regarding potential interaction mechanisms occurring between ponatinib ( Pntb ), a tyrosine kinase inhibitor of third‐generation and double‐stranded deoxyribonucleic acid (dsDNA). The aim of these studies was to elucidate the mechanism underlying the interaction between Pntb and DNA. This understanding could not only shed light on the changes occurring within living organisms but also, in the future, facilitate the development of new drugs for cancer treatment and improve their therapeutic effectiveness. The interactions were examined using square wave voltammetry (SWV) and UV–Vis spectroscopy. In order to conduct voltammetric experiments, a glassy carbon electrode was used. The addition of dsDNA to ponatinib solution resulted in a decrease of current peaks of the latter and resulted in a positive peak potential shift, suggesting that the intercalative type of interaction is the most likely to occur. Studies with utilization of UV–Vis spectroscopy have revealed a hypochromic effect, resulting in lack of wavelength shift in absorption maximum, following the addition of DNA to ponatinib solution, and confirming that intercalation is the predominant interaction between Pntb and dsDNA. The calculated value of the binding constant for Pntb ‐dsDNA determined by SWV was equal to 1.71 × 10 7 M −1 . The values calculated from UV–Vis spectroscopy, however, were 1.13 × 10 7 M −1 .
Journals
2025 EN
Duša Filip · Šalplachta Jiří · Horká Marie
+4 more
ABSTRACT Timely identification of highly pathogenic bacteria is crucial for efficient mitigation of the connected harmful health effects. Matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry (MALDI‐TOF MS) of intact cells enables fast identification of the microorganisms based on their mass spectrometry protein fingerprint profiles. However, the MALDI‐TOF MS examination must be preceded by a time‐demanding cultivation of the native bacteria to isolate representative cell samples to obtain indicative fingerprints. Isoelectric focusing (IEF) is capable of separating bacterial cells according to their isoelectric point while effectively removing other non‐focusing compounds from sample matrix. In this work, we present a divergent‐flow IEF chip (DF‐IEF chip) fractionation as an alternative way for sample clean‐up and concentration of bacterial cells to prepare samples usable for following MALDI‐TOF MS analysis without the need of time‐demanding cultivation. By means of DF‐IEF chip method, we processed four species of highly pathogenic bacteria ( Bacillus anthracis , Brucella abortus , Burkholderia mallei , and Yersinia pestis ) inactivated with H 2 O 2 vapors or by heat treatment at 62.5°C for 24 h. The DF‐IEF chip method continually separated and concentrated the inactivated bacterial cells for subsequent detection using MALDI‐TOF MS. The content of the inactivated bacteria in the DF‐IEF chip fractions was evaluated with the MS analysis, where inactivated Y. pestis was found to be the most efficiently focusing species. Sensitivity analysis showed limits as low as 2 × 10 5 colony forming units per mL for inactivated B. anthracis .
Journals
2025 EN
Branicki Wojciech · PisarekPacek Aleksandra · Marszałek Kamila
+8 more
ABSTRACT The MC1R gene, which is responsible for most cases of red hair, affects other hair and skin colours and contributes to differences in pain sensitivity and consists of a single exon with a very high level of allelic heterogeneity. In this research, we show that the Oxford Nanopore Technology (ONT) offers a good alternative to study the MC1R sequence variation. MinION was used to sequence the 1590 bp MC1R exon and minimal promoter in a cohort of 126 subjects, including 65 red‐haired individuals, using the FLO‐MIN106 (R9.4) chemistry. Assigned DNA variants were validated using Ion Torrent technology provided with Ion Xpress Plus Fragment Library Kit and the Personal Genome Machine TM (PGM TM ). We show that the use of the latest sequencing kit V14 together with the FLO‐MIN114 (R10.4.1) flow cell has eliminated the systematic errors observed with the previous chemistry and allowed reliable detection of short indels important for phenotypic inference. Importantly, the use of the algorithm implemented in the EPI2ME software enabled convenient and accurate read‐based phase determination which can be useful in data interpretation.
Journals
2025 EN
Šejna Jakub · Šálek Vojtěch · Šulc Stanislav
+5 more
ABSTRACT This paper presents the results of a standard fire resistance test of a loaded steel beam in a horizontal furnace. The beam was tested in three configurations: (1) unprotected, (2) protected with a single 22 mm layer of oriented strand board, and (3) protected with a double layer of the same cladding. The study also describes the development of a model in Fire Dynamics Simulator to predict the thermal conditions in the furnace and to observe the temperature trends on the beam surface, on the cladding, and at various depths in the cladding. A comparison between calculated and measured temperatures showed good agreement for the unprotected beam. However, for the protected beams, the model underestimated temperatures after 15 and 30 min for the single‐layer and double‐layer protection, respectively. Several potential sources for the discrepancies are identified. The main reason lies probably in the model's inability to correctly account for the effect of gaps in the cladding joints. Future work will focus on improving the accuracy of the model by removing these identified limitations, with particular attention to the behavior of the cladding as a passive fire protection material.
Journals
2025 EN
Mizera Kamila · Sałasińska Kamila · Borucka Monika
+2 more
ABSTRACT The development of technology and newer requirements for materials leads to an increase in their production and storage. In the past, serious fires have occurred in public buildings, residential buildings, industrial halls, and warehouses where plastics were used or produced, resulting in disastrous consequences for the environment and human health. For this reason, it is important to examine the risks to people and the environment that arise during a fire in places where these materials are located and stored. Polyurethane foams (PUF) used in building insulation and the automotive industry have been analyzed to determine their flammability and smoke emission during combustion. The thermal stability of PUFs was assessed using simultaneous thermal analysis (STA). Released gases were identified using STA combined with FT‐IR (STA/FT‐IR). Fire resistance and smoke emission during combustion were evaluated using cone calorimetry and a smoke chamber. Differences in thermal decomposition and combustion characteristics, including smoke release, were observed. The combustion of semi‐rigid foam was accompanied by the lowest total smoke release and the lowest total heat release. However, the combustion of flexible foam was characterized by the highest amount of smoke and a high rate of heat release, despite only a 5% weight loss at the highest temperature. In the case of rigid foam, a large residue in the form of a carbonized layer was observed.
Journals
2025 EN
Vodicka Prokop · Janikova Andrea · Belada David
+12 more
ABSTRACT Primary central nervous system lymphomas (PCNSL) are rare malignancies with poor survival outcomes. The IELSG32 trial demonstrated efficacy of MATRix chemoimmunotherapy followed by autologous stem cell transplantation (auto‐SCT) in PCNSL patients aged ≤ 70 years with a performance status (PS) ECOG ≤ 3. However, long‐term real‐world results of MATRix/auto‐SCT therapy remain limited. This analysis, with a median follow‐up of 52 months, aimed to evaluate the outcomes of MATRix‐treated PCNSL patients in clinical practice. From 2015 to 2022, 280 PCNSL patients who received systemic therapy were identified in the NiHiL project (NCT03199066). Eighty‐eight individuals treated with MATRix entered the analysis. Endpoints included efficacy and safety of induction and consolidation therapy. Seventy‐eight patients who met key IELSG32 inclusion criteria (age ≤ 65 years and PS ECOG ≤ 3, or age 66–70 years and PS ECOG ≤ 2) achieved an overall response rate of 82% (complete remission rate 58%) following MATRix regimen. After median follow‐up of 52 months, 4‐year progression‐free survival and overall survival (OS) rates were 53% and 55%, respectively. Forty‐six (59%) patients completed MATRix treatment, and 32 (41%) discontinued induction therapy (15 toxicity, 11 infections, 5 progressive diseases, 1 refusal). The treatment‐related mortality was 8%. Among 67 patients with responsive/stable disease, 50 underwent consolidation with whole‐brain radiotherapy (WBRT, n = 13) or auto‐SCT ( n = 37). No significant survival differences were observed between WBRT and auto‐SCT (4‐year OS 84% vs. 74%, HR 0.61, 95% CI 0.16–2.29, p = 0.467). Long‐term real‐world outcomes of MATRix/auto‐SCT therapy are comparable to IELSG32, supporting its use in younger, fit PCNSL patients. Clinical Trial Registration The data in this analysis were collected in the observational Czech non‐Hodgkin lymphoma registry “NiHiL” (NCT03199066)