Microbial transglutaminase (MTG) is an enzyme widely used in the food industry because it creates cross-links between proteins, enhancing the texture and stability of food products. Its unique properties make it a valuable tool for modifying the functional characteristics of proteins, significantly impacting the quality and innovation of food products. In this study, response surface methodology was employed to optimize the fermentation conditions for microbial transglutaminase production by the strain Streptoverticillium cinnamoneum KKP 1658. The effects of nitrogen dose, cultivation time, and initial pH on the activity of the produced transglutaminase were investigated. The significance of the examined factors was determined as follows: cultivation time > nitrogen dose > pH. The interaction between nitrogen dose and cultivation time was found to be crucial, having the second most significant impact on transglutaminase activity. Optimal conditions were identified as 48 h of cultivation with a 2% nitrogen source dose and an initial medium pH of approximately 6.0. Under these conditions, transglutaminase activity ranged from 4.5 to 5.5 U/mL. The results of this study demonstrated that response surface methodology is a promising approach for optimizing microbial transglutaminase production. Future applications of transglutaminase include the development of modern food products with improved texture and nutritional value, as well as its potential use in regenerative medicine for creating biomaterials and tissue scaffolds. This topic is particularly important and timely as it addresses the growing demand for innovative and sustainable solutions in the food and biomedical industries, contributing to an improved quality of life.

• MeSH
• dusík * metabolismus   MeSH
• fermentace * MeSH
• koncentrace vodíkových iontů MeSH
• kultivační média * chemie   MeSH
• transglutaminasy * metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH

The alveolar-capillary interface is the key functional element of gas exchange in the human lung, and disruptions to this interface can lead to significant medical complications. However, it is currently challenging to adequately model this interface in vitro, as it requires not only the co-culture of human alveolar epithelial and endothelial cells but mainly the preparation of a biocompatible scaffold that mimics the basement membrane. This scaffold should support cell seeding from both sides, and maintain optimal cell adhesion, growth, and differentiation conditions. Our study investigates the use of polycaprolactone (PCL) nanofibers as a versatile substrate for such cell cultures, aiming to model the alveolar-capillary interface more accurately. We optimized nanofiber production parameters, utilized polyamide mesh UHELON as a mechanical support for scaffold handling, and created 3D-printed inserts for specialized co-cultures. Our findings confirm that PCL nanofibrous scaffolds are manageable and support the co-culture of diverse cell types, effectively enabling cell attachment, proliferation, and differentiation. Our research establishes a proof-of-concept model for the alveolar-capillary interface, offering significant potential for enhancing cell-based testing and advancing tissue-engineering applications that require specific nanofibrous matrices.

• MeSH
• bazální membrána MeSH
• lidé MeSH
• nanovlákna * chemie   MeSH
• ověření koncepční studie MeSH
• plicní alveoly * chemie   cytologie   MeSH
• polyestery * chemie   MeSH
• tkáňové inženýrství * metody   MeSH
• tkáňové podpůrné struktury * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

BACKGROUND: The primary objective of this study was to conduct a comprehensive questionnaire survey on the practices of research ethics committees reviewing academic research projects in Czechia. The study aims to provide an unbiased and objective assessment of the current practices of research ethics committees, namely to obtain the missing data on their functioning in the context of academic research, to identify difficulties and shortages that threaten the responsible functioning of research ethics committees in the country and to investigate the implementation of Additional Protocol on Biomedical Research CETS No. 195 in their practice. Such research has never been conducted in Czechia. METHODS: This was a mixed-methods study, in which the online survey with closed and open-ended questions was chosen to explore the situation regarding ethics assessment of research involving human participants. We developed a questionnaire containing 18 questions concerning several aspects of the functioning of research ethics committees. The questionnaire was in Czech language and was administered through the Qualtrics platform anonymously. The target group of 61 research ethics committees at research institutions was approached by emails and we received 43 completely filled questionnaires, i.e., response rate of 67%. RESULTS: We obtained valuable data on the functioning of research ethics committees in Czechia in three main domains: the mandate and composition of the committee; the scope of its agenda; the process of evaluation including the voting procedure. In addition, the final set of open-ended questions provided an in-depth look at the problems faced by research ethics committees in Czechia. From the results is evident that the responsible assessment of the ethics of research involving human subjects is still not satisfactorily addressed and established for routine practice in the country. CONCLUSIONS: The outcomes of our study revealed that the main problem of research ethics in Czechia is the lack of national legislation on research ethics governance. To address this problem, the country requires a legislative framework accompanied by supportive measures aimed at educating, guiding and advising research ethics committees, especially in the Czech academic environment. TRIAL REGISTRATION NUMBER: Not applicable.

• MeSH
• biomedicínský výzkum etika   MeSH
• etické komise - výzkum * MeSH
• etické zhodnocení MeSH
• etika výzkumu * MeSH
• experimenty na lidech etika   MeSH
• lidé MeSH
• průzkumy a dotazníky MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH

Microflow liquid chromatography interfaced with mass spectrometry (μLC-MS/MS) is increasingly applied for high-throughput profiling of biological samples and has been proven to have an acceptable trade-off between sensitivity and reproducibility. However, lipidomics applications are scarce. We optimized a μLC-MS/MS system utilizing a 1 mm inner diameter × 100 mm column coupled to a triple quadrupole mass spectrometer to establish a sensitive, high-throughput, and robust single-shot lipidomics workflow. Compared to conventional lipidomics methods, we achieve a ∼4-fold increase in response, facilitating quantification of 351 lipid species from a single iPSC-derived cerebral organoid during a 15 min LC-MS analysis. Consecutively, we injected 303 samples over ∼75 h to prove the robustness and reproducibility of the microflow separation. As a proof of concept, μLC-MS/MS analysis of Alzheimer's disease patient-derived iPSC cerebral organoid reveals differential lipid metabolism depending on APOE phenotype (E3/3 vs E4/4). Microflow separation proves to be an environmentally friendly and cost-effective method as it reduces the consumption of harmful solvents. Also, the data demonstrate robust, in-depth, high-throughput performance to enable routine clinical or biomedical applications.

• MeSH
• apolipoproteiny E MeSH
• chromatografie kapalinová metody   MeSH
• fenotyp MeSH
• kapalinová chromatografie-hmotnostní spektrometrie * MeSH
• lidé MeSH
• lipidomika MeSH
• reprodukovatelnost výsledků MeSH
• tandemová hmotnostní spektrometrie * metody   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Nanoparticles have drawn significant interest in a range of applications, ranging from biomedical to environmental sciences, due to their distinctive physicochemical characteristics. In this study, it was reported that simple biological production of Ag, Se, and bimetallic Ag2Se nanoparticles (NPs) with Pseudomonas aeruginosa is a promising, low-cost, and environmentally friendly method. For the first time in the scientific literature, Ag2Se nanoparticles have been generated via green bacterial biosynthesis. UV-vis spectroscopy, X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), transmission electron microscopy (TEM), scanning electron microscopy (SEM), and EDX were used to characterize the produced NPs. Biosynthesized NPs were examined for antibacterial, antibiofilm, and photocatalytic properties, and it was determined that the effects of NPs were dose dependent. The biosynthesized AgNPs, SeNPs, and Ag2Se NPs showed anti-microbial activity against Escherichia coli and Staphylococcus aureus. Minimal inhibitory concentrations (MICs) of E. coli and S. aureus were between 150 and 250 μg/mL. The NPs showed antibiofilm activity against E. coli and S. aureus at sub-MIC levels and reduced biofilm formation by at least 80% at a concentration of 200 μg/mL of each NPs. To photocatalyze the breakdown of Congo red, Ag, Se, and Ag2Se NPs were utilized, and their photocatalytic activity was tested at various concentrations and intervals. A minor decrease of photocatalytic degradation was detected throughout the NPs reuse operation (five cycles). Based on the encouraging findings, the synthesized NPs demonstrated antibacterial, antibiofilm, and photocatalytic properties, suggesting that they might be used in pharmaceutical, medical, environmental, and other applications.

• MeSH
• antibakteriální látky * farmakologie   chemie   chemická syntéza   MeSH
• biofilmy * účinky léků   MeSH
• Escherichia coli * účinky léků   MeSH
• katalýza MeSH
• kovové nanočástice * chemie   MeSH
• mikrobiální testy citlivosti * MeSH
• Pseudomonas aeruginosa * účinky léků   metabolismus   MeSH
• selen chemie   farmakologie   MeSH
• sloučeniny stříbra chemie   farmakologie   MeSH
• Staphylococcus aureus * účinky léků   MeSH
• stříbro * chemie   farmakologie   metabolismus   MeSH
• technologie zelené chemie * MeSH
• Publikační typ
• časopisecké články MeSH

The review focuses on the design of detection cells, the use of microcontrollers for processing and evaluation of the detection signal, and the development of multi-detection systems for electromigration, liquid chromatography, flow-through and microfluidic techniques. A separate section is the introduction of modern 3D printing techniques and the use of new printing materials for the design of multidetection systems. In addition to traditional utilisation in separation techniques, new versions of contactless conductivity detectors are finding applications in FIA, SIA, portable and paper based analytical systems or as independent sensors. Applicationwise, C4Ds find new use in gas detection, segmented flow monitoring, as part of point of care devices, and in many other biomedical, environmental, agricultural and industrial applications.

• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

Chimeric antigen receptor (CAR) T-cell is a cutting edge technology for targeted cell therapy of oncologic diseases. Promising clinical results were reported for hematological malignancies, but the results in solid tumors are not that encouranging yet. Here we propose to validate protocols for the production of CAR T-cells against solid tumor antigens under cGMP rules. We will focus mainly on target antigens GD2, PSMA, and PSCA. Standard operation protocols and analytical certificates will be presented to the State Institute for Drug Control for their approval. The consortium of three prominent research facilities will participate on this project: (i) International Clinical Research Center of St. Anne's University Hospital Brno (FNUSA-ICRC), (ii) Centre for Biomedical Image Analysis at Masaryk University Brno (MU-CBIA), and (iii) Institute of Hematology and Blood Transfusion in Prague (UHKT). Our main aim is to establish production of CAR T-cells for anti-solid tumor therapy which can be translated into clinical applications.

T lymfocyty s chimerickým antigenním receptorem (CAR) představují nejmodernější technologii v cílené buněčné terapii onkologických onemocnění. Slibné klinické výsledky byly publikovány v léčbě hemato-onkologických malignit, avšak výsledky v léčbě solidních nádorů nejsou zatím tak povzbudivé. V navrhovaném projektu se budeme věnovat validaci protokolů pro výrobu CAR T-lymfocytů proti solidním nádorům v režimu správné laboratorní praxe. Zaměříme se hlavně na cílové antigeny GD2, PSMA a PSCA. Standardní operační protokoly a analytické certifikáty budou předány Státnímu ústavu pro kontrolu léčiv ke schválení. Na projektu budou spolupracovat tři špičková výzkumná pracoviště: (i) Mezinárodní centrum klinického výzkumu Fakultní nemocnice u sv. Anny v Brně, (ii) Centrum analýzy biomedicínského obrazu na Masarykově Univerzitě v Brně (MU-CBIA) a (iii) Ústav hematologie a krevní transfuze v Praze (ÚHKT). Naším hlavním cílem je zavést technologii výroby CAR T-lymfocytů pro cílenou buněčnou terapii solidních tumorů a tím umožnit přenos do klinické praxe.

• Klíčová slova
• advanced therapy medicinal products, solid tumors, T-lymfocyty, T-cells, solidní tumory, Chimerický antigenní receptor, Správná laboratorní praxe, Přípravky moderní terapie, Chimeric antigen receptor, Current Good Manufacturing Practice,

• NLK Publikační typ
• závěrečné zprávy o řešení grantu AZV MZ ČR

Restoring the structures and functions of tissues along with organs in human bodies is a topic gathering attention nowadays. These issues are widely discussed in the context of regenerative medicine. Excipients/delivery systems play a key role in this topic, guaranteeing a positive impact on the effectiveness of the drugs or therapeutic substances supplied. Advances in materials engineering, particularly in the development of hydrogel biomaterials, have influenced the idea of creating an innovative material that could serve as a carrier for active substances while ensuring biocompatibility and meeting all the stringent requirements imposed on medical materials. This work presents the preparation of a natural polymeric material based on pullulan modified with silymarin, which belongs to the group of flavonoids and derives from a plant called Silybum marianum. Under UV light, matrices with a previously prepared composition were crosslinked. Before proceeding to the next stage of the research, the purity of the composition of the matrices was checked using Fourier-transform infrared (FT-IR) spectroscopy. Incubation tests lasting 19 days were carried out using incubation fluids such as simulated body fluid (SBF), Ringer's solution, and artificial saliva. Changes in pH, electrolytic conductivity, and weight were observed and then used to determine the sorption capacity. During incubation, SBF proved to be the most stable fluid, with a pH level of 7.6-7.8. Sorption tests showed a high sorption capacity of samples incubated in both Ringer's solution and artificial saliva (approximately 350%) and SBF (approximately 300%). After incubation, the surface morphology was analyzed using an optical microscope for samples demonstrating the greatest changes over time. The active substance, silymarin, was released using a water bath, and then the antioxidant capacity was determined using the Folin-Ciocâlteu test. The tests carried out proved that the material produced is active and harmless, which was shown by the incubation analysis. The continuous release of the active ingredient increases the biological value of the biomaterial. The material requires further research, including a more detailed assessment of its balance; however, it demonstrates promising potential for further experiments.

• MeSH
• glukany * chemie   MeSH
• koncentrace vodíkových iontů MeSH
• lékové transportní systémy metody   MeSH
• lidé MeSH
• nosiče léků * chemie   MeSH
• polyethylenglykoly * chemie   MeSH
• silymarin * chemie   MeSH
• spektroskopie infračervená s Fourierovou transformací MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Herein, the recent advances in the development of resorbable polymeric-based biomaterials, their geometrical forms, resorption mechanisms, and their capabilities in various biomedical applications are critically reviewed. A comprehensive discussion of the engineering approaches for the fabrication of polymeric resorbable scaffolds for tissue engineering, drug delivery, surgical, cardiological, aesthetical, dental and cardiovascular applications, are also explained. Furthermore, to understand the internal structures of resorbable scaffolds, representative studies of their evaluation by medical imaging techniques, e.g., cardiac computer tomography, are succinctly highlighted. This approach provides crucial clinical insights which help to improve the materials' suitable and viable characteristics for them to meet the highly restrictive medical requirements. Finally, the aspects of the legal regulations and the associated challenges in translating research into desirable clinical and marketable materials of polymeric-based formulations, are presented.

• MeSH
• biokompatibilní materiály chemie   MeSH
• lékové transportní systémy * metody   MeSH
• lidé MeSH
• polymery * chemie   MeSH
• tkáňové inženýrství * metody   MeSH
• tkáňové podpůrné struktury chemie   MeSH
• vstřebatelné implantáty MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

INTRODUCTION: The methods for diagnosing compartment syndrome non-invasively remain under debate. Bioimpedance measurements offer a promising avenue in clinical practice, detecting subtle changes in organ impedance due to volume shifts. This study explores bioimpedance measurement as a novel, painless method for diagnosing compartment syndrome, potentially enabling continuous monitoring. OBJECTIVE: This work aims to develop a prototype device for non-invasive diagnosis of compartment syndrome based on bioimpedance changes and assess initial results through in vitro experiments on inanimate biological material. We assume a change in the bioimpedance value after the application of physiological solution. MATERIALS AND METHODS: Between 2018 and 2022, a prototype device for diagnosing limb compartment syndrome was collaboratively developed with the Department of Cybernetics and Biomedical Engineering at the Technical University of Ostrava, Czech Republic. This device operates by comparing bioimpedance between two compartments, one of which is pathologically affected (experiencing compartment syndrome). The Bioimpedance Analyzer for Compartment Syndrome (BACS) has been utilized to conduct measurements on inanimate biological material in laboratory settings. Two samples of duck and chicken tissue, as well as piglets, were employed for these experiments. According to the size of sample was compartment syndrome simulated by injecting 20-120 mL saline into one limb (breast) while leaving the other as a control. Invasive intramuscular pressure measurements were conducted post-saline injection using a conventional device (Stryker). Changes in bioimpedance were evaluated following saline application. RESULTS: The non-invasive bioimpedance measurement instrument has been developed. It meets the safety requirements of European standard EN 60601-1. Measurement of accuracy showed minimal deviation for both channels (1.08% for the left channel and 1.84% for the right channel) when measuring on resistors. Ten measurements were conducted using the BACS prototype - two on chicken legs, two on duck breasts, two on duck legs, and four on piglets. Compartment syndrome simulation was achieved for all 10 measurements (IMP variance 31-45 mmHg). Following saline application, a notable decrease in bioimpedance was observed in the compartment simulating compartment syndrome (decrease by 12-78 Ω). CONCLUSION: Non-invasive methods could revolutionize limb compartment syndrome diagnosis, offering advantages such as non-invasiveness and continuous monitoring of compartment swelling.

• Publikační typ
• časopisecké články MeSH