Cíl: Minireview podává přehled aktuálního stavu měření 25-hydroxyvitaminu D v krevním séru. Metoda: Hodnocení analytické kvality měření 25-hydroxyvitaminu v programech externího hodnocení kvality. Výsledky separačních a imunochemických metod, stav metrologické reference a standardizace, předměty měření (analyty), preanalytická stabilita. Závěr: Je nutná standardizace měření jako podmínka vyšší úrovně srovnatelnosti mezi metodami a mezi laboratořemi využitím a rozvojem referenční metodiky a aplikací již existujících referenčních materiálů.
Objective: A mini-review on the current state in serum 25-hydroxyvitamin D measurement. Methods: The review of used routine methods for the measurement the 25-hydroxyvitamin D in serum. The assessment of analytical quality by means of inter-laboratory comparison programs is presented. The current state of metrological reference and standardization is outlined. The methods, based on separation, with immunochemistry methods are compared. Problem of suitable analytes as subjects of measurement. Stability of analyte is described. Conclusion: There is the necessity of standardization for achieving the interlaboratory comparability and clinical utility by means of using reference material and reference method in routine practice.
- Keywords
- srovnatelnost metod, LC-MS/MS, metrologické reference,
- MeSH
- 25-Hydroxyvitamin D 2 analysis diagnostic use MeSH
- Immunochemistry methods MeSH
- Plasma chemistry MeSH
- Specimen Handling standards MeSH
- Reference Values MeSH
- Reference Standards MeSH
- Serum chemistry MeSH
- Laboratory Proficiency Testing utilization MeSH
- Publication type
- Review MeSH
Závěrečná zpráva o řešení grantu Interní grantové agentury MZ ČR
52 l. : il., tab. ; 31 cm
Vyvinutí a ověření nové metody stanovení cirkulujících hladin endogenních hydroxylovaných metabolitů vitaminu D pomocí imunochemických a chromatografických technik s UV nebo fluorescenční detekcí nativních nebo derivatizovaných analytů. Stanovení cirkulujících hydroxylovaných metabolitů vitaminu D u dětí trpících závažnou ledvinnou nedostatečností a u pacientů s poruchou remodelace kosti.; Developing and validation of method for determination of circulating endogenous vitamin D hydroxylated metabolites using immunochemical and chromatographic techniques with UV or fluorimetric detection of native or derivatized analytes. Determination of circulating vitamin D hydroxylated metabolites in children with serious renal failure (200 sample per year) and in patients with defective bone remodeling (200 samples).
- MeSH
- Chromatography methods utilization MeSH
- Fluorometry MeSH
- Immunoassay MeSH
- Immunochemistry MeSH
- Specimen Handling MeSH
- Vitamin D metabolism MeSH
- Conspectus
- Biochemie. Molekulární biologie. Biofyzika
- NML Fields
- biochemie
- vnitřní lékařství
- NML Publication type
- závěrečné zprávy o řešení grantu IGA MZ ČR
Oxime cholinesterase reactivators (oximes) are used to counteract organophosphate intoxication. Charged oximes are administered via intramuscular or intravenous injection when the majority of dose is unmetabolized and is excreted as urine. In this study, the effects of selected double charged oximes were determined in the HK-2 cell line as a model for renal toxicity screening. Some effects on dehydrogenase activity were found for obidoxime, asoxime (syn. HI-6), K027, and K203. The effects of K868 and K869 were found to be unreliable due to rapid degradation of both chlorinated oximes in the assay medium, resulting for K868 in an isoxazole-pyridinium product.
- MeSH
- Cell Line MeSH
- Kidney drug effects metabolism MeSH
- Humans MeSH
- Molecular Structure MeSH
- Oximes administration & dosage adverse effects chemistry MeSH
- Cholinesterase Reactivators administration & dosage adverse effects chemistry MeSH
- Dose-Response Relationship, Drug MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
Alzheimer's disease (AD) is a multifaceted neurodegenerative disorder for which current treatments provide only symptomatic relief, primarily through cholinesterase (ChE) inhibition and N-methyl-d-aspartate receptor (NMDAR) antagonism. To improve therapeutic efficacy and safety, we designed and synthesized 16 novel tacrine derivatives modified at position 7 with various (hetero)aryl groups or deuterium substitution. Initially, in silico screening predicted favorable CNS permeability and oral bioavailability. Subsequent in vitro evaluations demonstrated significant inhibitory potency against acetylcholinesterase (AChE) and butyrylcholinesterase (BChE), with derivatives 5i and 5m displaying particularly promising profiles. Metabolic stability assessed using human liver microsomes revealed enhanced stability for compound 5e, whereas 5i and 5m underwent rapid metabolism. Notably, compound 7 showed improved metabolic stability attributed to deuterium incorporation. The newly synthesized compounds were further tested for antagonistic activity on the GluN1/GluN2B subtype of NMDAR, with compound 5m exhibiting the most potent and voltage-independent inhibition. The ability of these compounds to permeate the blood-brain barrier (BBB) was confirmed through in vitro PAMPA assays. In preliminary hepatotoxicity screening (HepG2 cells), most derivatives exhibited higher cytotoxicity than tacrine, emphasizing the ongoing challenge in hepatotoxicity management. Based on its overall favorable profile, compound 5m advanced to in vivo pharmacokinetic studies in mice, demonstrating efficient CNS penetration, with brain concentrations exceeding plasma levels (brain-to-plasma ratio 2.36), indicating active transport across the BBB. These findings highlight compound 5m as a promising tacrine-based multi-target-directed ligand, supporting further preclinical development as a potential therapeutic candidate for AD.
- MeSH
- Acetylcholinesterase metabolism MeSH
- Alzheimer Disease * drug therapy metabolism MeSH
- Biological Availability MeSH
- Butyrylcholinesterase metabolism MeSH
- Cholinesterase Inhibitors * pharmacology chemistry chemical synthesis MeSH
- Blood-Brain Barrier metabolism MeSH
- Microsomes, Liver metabolism MeSH
- Humans MeSH
- Ligands MeSH
- Molecular Structure MeSH
- Mice MeSH
- Receptors, N-Methyl-D-Aspartate * antagonists & inhibitors metabolism MeSH
- Tacrine * pharmacology chemistry chemical synthesis MeSH
- Dose-Response Relationship, Drug MeSH
- Structure-Activity Relationship MeSH
- Animals MeSH
- Check Tag
- Humans MeSH
- Male MeSH
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Keywords
- imobilní pacient, škola zad,
- MeSH
- Back Pain MeSH
- Physical Exertion MeSH
- Nurses MeSH
- Moving and Lifting Patients MeSH
Diabetic cardiomyopathy is the leading cause of mortality in type 1 diabetes. Thus study of cardiomyocyte morphology and function during early stages of diabetes using modern analytical methods is of critical importance. Therefore, using confocal microscopy, we determined metric parameters, volumes and contractility, with calcium transients in isolated left-ventricular myocytes at one week after induction of diabetes in rats. Myocyte volume analysis from 3D confocal scans was performed using an automated contour detection algorithm that took the actual shape of the myocytes into account. We showed a significant reduction in myocyte volume in diabetic animals. We also showed a significant reduction in length and width but not in thickness of the myocytes, which suggests disproportional reorganization of the structure of the heart tissue during short-term diabetes. From a functional point of view, we observed a significant decrease in cell shortening at a stimulation frequency of 0.5 Hz. This was accompanied by a decrease in calcium transient amplitude. Together, these data suggest that impaired calcium handling is one of the factors that contributes to the observed decrease in myocyte shortening during early stages of streptozotocin-induced diabetes in rats.
- MeSH
- Algorithms MeSH
- Time Factors MeSH
- Diabetic Cardiomyopathies etiology metabolism pathology physiopathology MeSH
- Electric Stimulation MeSH
- Diabetes Mellitus, Experimental chemically induced complications MeSH
- Image Interpretation, Computer-Assisted MeSH
- Myocytes, Cardiac metabolism pathology MeSH
- Microscopy, Confocal MeSH
- Myocardial Contraction * MeSH
- Rats MeSH
- Rats, Wistar MeSH
- Streptozocin MeSH
- Cell Shape * MeSH
- Calcium Signaling MeSH
- Animals MeSH
- Check Tag
- Rats MeSH
- Male MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
Enzyme kinetic measurements are important for the characterization and engineering of biocatalysts, with applications in a wide range of research fields. The measurement of initial reaction velocity is usually slow and laborious, which motivated us to explore the possibilities for automating this process. Our model enzyme is the maize β-glucosidase Zm-p60.1. Zm-p60.1 plays a significant role in plant growth and development by regulating levels of the active plant hormone cytokinin. Zm-p60.1 belongs to a wide group of hydrolytic enzymes. Members of this group hydrolyze several different types of glucosides, releasing glucose as a secondary product. Enzyme kinetic measurements using artificial substrates are well established, but burdensome and time-consuming. Thus, they are a suitable target for process automation. Simple optical methods for enzyme kinetic measurements using natural substrates are often impossible given the optical properties of the enzymatic reaction products. However, we have developed an automated method based on glucose detection, as glucose is released from all substrates of glucosidase reactions. The presented method can obtain 24 data points from up to 15 substrate concentrations to precisely describe the enzyme kinetics. The combination of an automated liquid handling process with assays that have been optimized for measuring the initial hydrolysis velocity of β-glucosidases yields two distinct methods that are faster, cheaper, and more accurate than the established protocols.
- MeSH
- Automation MeSH
- beta-Glucosidase chemistry MeSH
- Catalysis MeSH
- Kinetics MeSH
- Zea mays enzymology MeSH
- Plant Proteins chemistry MeSH
- Publication type
- Journal Article MeSH
OBJECTIVE: Reporting epidemiological data on prehospital cardiac arrest in the Pilsen Region in 2022. Expression of cardiopulmonary resuscitation success using the Cerebral Performance Category (CPC) score. MATERIALS AND METHODS: The study looked at the survival rate of out-of-hospital sudden cardiac arrest in all patients in whom emergency medical services performed cardiopulmonary resuscitation (CPR). The study covered the period from 1 January 2022 to 31 December 2022. Both electronic and paper medical records were used to obtain data. All cases were evaluated according to Utstein-style guidelines. RESULTS: During the studied period, emergency response teams in the Pilsen Region carried out CPR in 499 cases. The incidence of prehospital CPR was 88.43 cases per 100,000 population. A total of 146 patients (29.26%) were referred to the hospital with spontaneous circulation, and results indicating survival with a good neurological outcome of CPC 1 or 2 were recorded in 48 cases (9.62%). The first monitored rhythm was shockable in 119 cases (23.85%). In this subgroup, ROSC was achieved in 71 cases (59.66%) and 61 of them (51.26%) were referred to hospital. In this study subgroup, a total of 36 patients (30.25%) achieved a good neurological outcome with a CPC score of 1 or 2. CONCLUSION: The study presented epidemiological data on OHCA and prehospital CPR in the Pilsen region in 2022. The data obtained shows a survival rate with good neurological outcome in 9.62% of cases.
- MeSH
- Cardiopulmonary Resuscitation * methods MeSH
- Humans MeSH
- Hospitals MeSH
- Registries MeSH
- Emergency Medical Services * methods MeSH
- Out-of-Hospital Cardiac Arrest * epidemiology therapy MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
