Cíl studie: Cílem této publikace bylo popsat výskyt maligní hypertermie (MH) v české a slovenské populaci, včetně genetických výsledků a důvodů hlášení do MH diagnostického centra. Typ studie: Retrospektivní observační kohortová studie. Typ pracoviště: Akademické centrum maligní hypertermie Masarykovy univerzity. Materiál a metody: Zkoumáni byli všichni probandi referovaní do našeho diagnostického centra MH od roku 2002 do 2022. Každý proband byl zástupcem jedné nepříbuzné rodiny. Probandi byli vyšetřováni dle diagnostických doporučení aktuálně platných v době hlášení prostřednictvím molekulárně genetického vyšetření a svalového in vitro kontrakčního testu (IVCT). Do výsledné analýzy byli zařazeni všichni probandi s ukončenou MH diagnostikou. Výsledky: Do konce 2022 bylo zaznamenáno v souvislosti s rizikem MH celkem 303 hlášení. Bylo provedeno 236 IVCT testů, 129 negativních a 107 pozitivních. Celkem je v našem souboru 223 osob vnímavých k MH (MHS) ze 75 českých a 8 slovenských rodin. Diagnostická patogenní varianta v genu RYR1 byla nalezena pouze u 37 MHS rodin (45 %). U 87 rodin byla MH vyloučena (MHN). Závěr: Za 20 let systematické práce je evidován v naší české a slovenské populaci unikátní soubor 83 MHS rodin. I přes rutinní zpřístupnění nejmodernější diagnostiky MH více než polovina našich MHS rodin nemá potvrzen genetický podklad MH, a může profitovat z vědeckého pokroku a dalšího výzkumu.
Objective: The aim of this publication was to describe the occurrence of malignant hyperthermia (MH) in the Czech and Slovak populations, including genetic results and reasons for reporting to the MH diagnostic centre. Design: Retrospective observational cohort study. Setting: Academic Centre of Malignant Hyperthermia of Masaryk University. Material and methods: All probands referred to our MH diagnostic centre from 2002 to 2022 were enrolled. Each proband was a representative of one unrelated family. Probands were investigated according to diagnostic guidelines valid at the time of reporting by molecular genetic testing and muscle in vitro contraction test (IVCT). All probands with completed MH diagnostics were included in the final analysis. Results: By the end of 2022, there were a total of 303 reports related to the risk of MH. A total of 236 IVCT tests were performed, 129 negatives and 107 positives. There were 223 persons susceptible to MH (MHS) registered from 75 Czech and 8 Slovak families. A diagnostic pathogenic variant in the RYR1 gene was found only in 37 MHS families (45%). MH was excluded in 87 families (MHN). Conclusion: In 20 years of systematic work, a unique set of 83 MHS families has been collected in our Czech and Slovak populations. Despite the routine availability of state-of-the-art MH diagnostics, more than half of our MH families do not have a confirmed MH genetic background and can benefit from scientific progress and further research.
- MeSH
- Anesthesia adverse effects MeSH
- Dantrolene therapeutic use MeSH
- Epidemiologic Studies MeSH
- Genetic Techniques MeSH
- Muscle, Skeletal physiopathology drug effects MeSH
- Humans MeSH
- Malignant Hyperthermia * diagnosis epidemiology drug therapy genetics MeSH
- Ryanodine Receptor Calcium Release Channel genetics MeSH
- Muscle Contraction drug effects MeSH
- In Vitro Techniques methods statistics & numerical data MeSH
- High-Throughput Nucleotide Sequencing MeSH
- Check Tag
- Humans MeSH
- Publication type
- Research Support, Non-U.S. Gov't MeSH
- Geographicals
- Czech Republic MeSH
- Slovakia MeSH
Inhibitory Janusových kináz (JAK) patří mezi cílené malé molekuly uplatňující se při přenosu signálu z extracelulárního prostoru do jádra buňky. Díky své velikosti působí intracelulárně, jsou selektivní k cílům, ke kterým vykazují afinitu, a jejich účinnost mimo cíl může být zvýšena v závislosti na dávce. K aktivaci Janusových kináz dochází po navázání adenosintrifosfátu (ATP). Obsazením katalytického místa pro ATP inhibitorem JAK s vyšší afinitou, než jakou má ATP, je tak zabráněno vazbě ATP a kinázové aktivitě, přičemž u inhibitorů JAK je tato inhibice reverzibilní a částečná. Jelikož existují čtyři izoformy JAK (JAK1, JAK2, JAK3 a TYK2), jsou vyvíjeny inhibitory JAK s různými profily selektivity. Signální dráhy významné v případě atopické dermatitidy (AD) jsou charakteristické zejména přítomností JAK1. Upadacitinib je reverzibilní inhibitor JAK1, který vykazuje nejvyšší selektivitu pro JAK1 oproti jiným izoformám JAK v enzymatických i buněčných testech in vitro. Účinnost upadacitinibu v léčbě pacientů se středně závažnou až těžkou AD byla prokázána v rámci klinických studií III. fáze Measure Up 1 a Measure Up 2 se srovnatelným výskytem nežádoucích účinků oproti placebu. Z hlediska dosažení příznivého bezpečnostního profilu hraje důležitou úlohu právě selektivita inhibitorů JAK.
Janus kinase (JAK) inhibitors are targeted small molecules involved in signal transduction from the extracellular space to the cell nucleus. They act intracellularly, are selective to targets to which they show affinity, and their off-target efficacy may be increased in a dose- dependent manner. Activation of JAK happens when adenosine triphosphate (ATP) binds the ATP binding site in JAK enzymes. If this site is occupied by JAK inhibitor with a higher affinity than ATP, ATP binding and, therefore, kinase activity is prevented. Inhibition by JAK inhibitors is reversible and partial. As there are four isoforms of JAK (JAK1, JAK2, JAK3 and TYK2), JAK inhibitors with different selective profiles are being developed. Signaling pathways critical in atopic dermatitis (AD) are characterized by the presence of JAK1. Upadacitinib is a reversible JAK1 inhibitor showing the highest selectivity for JAK1 over other JAK isoforms in in vitro enzymatic and cellular assays. The efficacy of upadacitinib in the treatment of patients with moderate to severe AD has been demonstrated in phase III clinical trials Measure Up 1 and Measure Up 2 with a comparable incidence of adverse events compared to placebo. The selectivity of the JAK inhibitors plays an important role in achieving a good safety profile.
- MeSH
- Dermatitis, Atopic drug therapy MeSH
- Janus Kinase Inhibitors * administration & dosage pharmacology adverse effects MeSH
- Janus Kinases drug effects MeSH
- Clinical Trials, Phase III as Topic MeSH
- Humans MeSH
- Signal Transduction drug effects MeSH
- In Vitro Techniques statistics & numerical data MeSH
- Treatment Outcome MeSH
- Check Tag
- Humans MeSH
Polycyklické aromatické uhlovodíky (PAU) představují skupinu významných kontaminantů pracovního i životního prostředí. Při expozici PAU v pracovním prostředí se vedle inhalační expoziční cesty může významně uplatňovat i expozice dermální. Stávající experimentální údaje o intenzitě a rychlosti penetrace látek do systémové cirkulace jsou zatím omezené. Předkládaný článek se zabývá metodickou a interpretační problematikou transepidermální absorpce PAU in vitro. Byla sledována intenzita (Flux) a rychlost (Lag time) penetrace naftalenu, fenanthrenu, pyrenu a benzo[a]pyrenu přes epidermální membránu z prasečího boltce. Experiment byl prováděn ve vertikálních statických difuzních komůrkách dle Franze (n = 32) a jako rozpouštědlo byl použit isopropyl-myristát. Parametr Flux (nmol/cm2/hod) dosáhl hodnot 95,7 ± 45,5 u naftalenu, 19,5 ± 8,7 u fenanthrenu, 4,38 ± 1,98 u pyrenu a 0,21 ± 0,08 u benzo[a]pyrenu. Parametr Lag time (hod) hodnot 0,26 ± 0,17 u naftalenu, 2,12 ± 0,41 u fenanthrenu, 3,25 ± 0,50 u pyrenu a 11,2 ± 4,08 u benzo[a]pyrenu. Hodnota parametru Flux klesala s molární hmotností PAU, zatímco hodnota parametru Lag time s molární hmotností PAU stoupala. Množství PAU, které za daný časový úsek penetrovalo přes epidermální membránu, se pohybovalo mezi 0,24 % (benzo[a]pyren) až 0,84 % (fenanthren) aplikované dávky. Penetrace PAU přes epidermální membránu vykazovala, v porovnání s experimenty na plné kůži, nižší stupeň variability dat. Z výsledků vyplývá, že použití epidermální membrány by mohlo zpřesňovat jak odhad vnitřní dávky PAU po dermální expozici, tak i odhad souvisejícího zdravotního rizika v rámci konzervativního expozičního scénáře. Experimenty s epidermální membránou jsou však časově i experimentálně náročné, bez možnosti objektivní kontroly integrity epidermální membrány, což může vést k finanční náročnosti testování, ztrátám vzorků a v neposlední řadě i k nárůstu rozdílů hodnot dat, získaných v různých laboratořích.
Polycyclic aromatic hydrocarbons (PAHs) are a group of significant contaminants in the occupational and living environment. In addition to the inhalation route of exposure in the occupational environment, there is also significant the dermal route of exposure. Existing experimental data on the intensity and rate of penetration of these substances into systemic circulation are still limited. The presented paper is focused on methodological and interpretative issues of trans-epidermal absorption of PAHs in vitro. In this study, we assessed the intensity (Flux) and rate (Lag time) of penetration of naphthalene, phenanthrene, pyrene and benzo[a]pyrene through the epidermal membrane derived from a pig ear. The experiment was performed using static vertical Franz diffusion cells (n = 32) and isopropyl myristate was used as a solvent. Flux (nmol/cm2/hour) reached 95.7 ± 45.5 in the case of naphthalene, 19.5 ± 8.7 of phenanthrene, 4.38 ± 1.98 of pyrene and 0.21 ± 0.08 of benzo[a]pyrene. Lag time (hour) was 0.26 ± 0.17 in the case of naphthalene, 2.12 ± 0.41 of phenanthrene, 3.25 ± 0.50 of pyrene and 11.2 ± 4.08 of benzo[a]pyrene. The Flux value decreased with the molecular weight of PAHs, while the Lag time increased with the molecular weight of PAHs. The amount of PAHs that penetrated through the epidermal membrane in a given time period ranged between 0.24% (benzo[a]pyrene) and 0.84% (phenanthrene) of the applied dose. The penetration of PAHs through the epidermal membrane showed a lower degree of data variability compared to full thickness skin experiments. The results suggest that the use of the epidermal membrane could define more accurately both the estimation of the internal dose of PAHs after dermal exposure and the estimation of the associated health risk within a conservative exposure scenario. However, experiments with using the epidermal membrane are time consuming and experimentally demanding, with no option of an objective control of the integrity of the epidermal membrane, which can lead to costly testing, loss of samples and, finally, an increase in the differences in data values obtained in different laboratories.
- MeSH
- Epidermis anatomy & histology physiopathology MeSH
- Data Interpretation, Statistical MeSH
- Skin Absorption * physiology MeSH
- Models, Animal MeSH
- Polycyclic Aromatic Hydrocarbons * adverse effects MeSH
- Swine MeSH
- In Vitro Techniques * methods statistics & numerical data MeSH
- Ear Auricle MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- MeSH
- Anti-Infective Agents pharmacology MeSH
- Cell Death drug effects radiation effects MeSH
- Candida tropicalis drug effects radiation effects MeSH
- Catalysis * radiation effects MeSH
- Metal Nanoparticles * chemistry MeSH
- Surface-Active Agents MeSH
- Sterilization * trends MeSH
- In Vitro Techniques statistics & numerical data MeSH
- Titanium chemistry MeSH
- Cell Survival drug effects radiation effects MeSH
In this study we analyzed molecular mechanisms of antioxidative protection of thymol and thyme oil using differentiated PC12 cells, a widely accepted neuronal model. Thymol due to multiple functions is commonly used for clinical applications. However, its action on nervous tissue remains poorly understood. We evaluated the effect of 24 h incubation of the cells with thymol (100 and 400 μM) and equivalent content of thyme oil. Microsomal glutathione transferase 1 (Mgst1) is an important player in anti-oxidative protection because of its transferase and peroxidase activities. Since its expression decreases during aging, we also used stable transfected cells with downregulated Mgst1 (PC12_M). We analyzed cell viability, lipid peroxidation level, glutathione content and expression of key enzymes responsible for cellular reduced glutathione – catalytic subunit of γ-glutamylcysteine ligase and glutathione reductase. Whereas thymol and thyme oil improved antioxidant capacity of control cells, diminished protection was observed in PC12_M line. Increasing interest in natural dietary components has focused attention on plants used as a rich source of bioactive phytochemicals. However, currently available information on the safe amount for thyme oil using appears to be insufficient. Our results indicate that the thyme oil should be used with caution, especially by elderly people.
- MeSH
- Antioxidants * physiology chemistry MeSH
- PC12 Cells drug effects MeSH
- Glutathione physiology drug effects MeSH
- Glutathione Transferase MeSH
- Neurons drug effects MeSH
- Plant Oils * pharmacology chemistry MeSH
- Lipid Peroxidation drug effects MeSH
- Aging drug effects MeSH
- Statistics as Topic MeSH
- In Vitro Techniques statistics & numerical data MeSH
- Thymol * pharmacology MeSH
- Thymus Plant MeSH
- Cell Survival drug effects MeSH
- Publication type
- Evaluation Study MeSH
- Research Support, Non-U.S. Gov't MeSH
Quaternary ammonium salts (chloride, bromide and iodide; QUATs) with n-alkyl chain lengths between C8 and C18 have been synthesized under optimized experimental conditions. These compounds were tested in vitro for antimicrobial activity against representative bacterial strains (Staphylococcus aureus CIP 4.83, Enterococcus hirae CIP 5855, Pseudomonas aeruginosa CIP 82118, Escherichia coli CIP 53126, Mycobacterium smegmatis CIP 7326) and fungal species (Aspergillus niger ATCC 16404, Candida albicans IP 118079, Trichophyton interdigitale IP 146583). While these compounds showed moderate antifungal activity, several of them (particularly C14-I−) may be considered as highly potential antibacterial agents against S. aureus, E. hirae and E. coli with MIC values lower than that of commercial benzalkonium chloride and ciprofloxacin used as standards. The relationship between the lipophilicity and the antibacterial activity of the tested QUATs was quantified by a multiple linear regression method.
- MeSH
- Anti-Bacterial Agents pharmacology chemical synthesis chemistry MeSH
- Antifungal Agents pharmacology chemical synthesis chemistry MeSH
- Quaternary Ammonium Compounds * pharmacology chemical synthesis chemistry MeSH
- In Vitro Techniques statistics & numerical data MeSH
- Structure-Activity Relationship MeSH
- Publication type
- Research Support, Non-U.S. Gov't MeSH
Avidity is an important feature of antibodies associated with their pathogenic effects. Anticardiolipin antibodies (aCLs) are the most commonly examined antiphospholipid antibodies, however, their avidity has been investigated only marginally. The aim of the study was to compare the avidity of the antibodies specifically bound to the cardiolipin-coated microtitrate wells with those bound to the wells bearing no cardiolipin in various conditions. We analysed 22 serum samples with high, medium and low levels of aCL IgG. The avidity of aCL IgG was determined in serially diluted sera by the ELISA method in the presence of increasing urea concentrations (from 2 to 8 mol/L) as a chaotropic agent. The serum dilution 1:50 and 1:100 and the concentrations of urea 6 mol/L and 8 mol/L seemed to be suitable for the determination of aCL avidity. The avidity of antibodies specifically bound to the cardiolipin and those bound to the cardiolipin-free wells significantly differed in their avidity. The simultaneous determination of higher-avidity aCL and lower-avidity polyspecific antibodies, whose presence complicates the ELISA methods by an increase of the background signal, might limit the shortcomings of some ELISA methods.
- MeSH
- Antibody Affinity * immunology MeSH
- Antibodies, Anticardiolipin * immunology MeSH
- Autoimmune Diseases blood MeSH
- Adult MeSH
- Enzyme-Linked Immunosorbent Assay * MeSH
- Immunoglobulin G immunology MeSH
- Middle Aged MeSH
- Humans MeSH
- Urea immunology MeSH
- Aged MeSH
- Sensitivity and Specificity MeSH
- Serum immunology MeSH
- In Vitro Techniques statistics & numerical data MeSH
- Check Tag
- Adult MeSH
- Middle Aged MeSH
- Humans MeSH
- Male MeSH
- Aged MeSH
- Female MeSH
- Publication type
- Evaluation Study MeSH
- Research Support, Non-U.S. Gov't MeSH
Alzheimer disease (AD) is the most common cause of progressive dementia in the elderly population, with prevalence of 5% after 65 year of age and is increasing to about 30% in people over 85 year. AD is a neurodegenerative and incurable disease. Currently, three inhibitors of acetylcholinesterase (AChE), galantamine, donepezil and rivastigmine, and one inhibitor of N-methyl-d-aspartate (NMDA) receptor are available as drugs for amelioration of the disease. Demand to prepare drugs for the therapy providing at least relieve of symptoms remains. In this experiment, the ability of standards (donepezil, galantamine, huperzine A, tacrine and 7-methoxytacrine) and precursors used for synthesis of new AD drugs (l-tryptophan, pyridoxine B6, tryptamine, acridine, chinoline, isochinoline, indole, pyridine and piperidine) to inhibit AChE, BChE and BACE or to have the antioxidant properties were determined. The results were compared using statistical expression of the relationship between the performed tests. In this experiment, IC50 for every one method and every compound were found. Beside this, prediction of free energy in a link to ln(IC50) was assessed using in silico tests. This article focuses on possibility to find the most suitable chemical precursors to be used in the next development of drugs for AD
- MeSH
- Acridines pharmacology MeSH
- Alzheimer Disease * drug therapy MeSH
- Antioxidants pharmacology MeSH
- Aspartic Acid Endopeptidases antagonists & inhibitors MeSH
- Quinolines pharmacology MeSH
- Cholinesterase Inhibitors pharmacology MeSH
- Chemistry, Pharmaceutical * methods statistics & numerical data MeSH
- Indoles pharmacology MeSH
- Isoquinolines pharmacology MeSH
- Neurodegenerative Diseases drug therapy MeSH
- Piperidines pharmacology MeSH
- Computer Simulation MeSH
- Pyridines pharmacology MeSH
- Pyridoxine pharmacology MeSH
- Amyloid Precursor Protein Secretases antagonists & inhibitors MeSH
- In Vitro Techniques statistics & numerical data MeSH
- Tryptamines pharmacology MeSH
- Tryptophan pharmacology MeSH
- Publication type
- Evaluation Study MeSH
- Research Support, Non-U.S. Gov't MeSH
Epigallocatechin-3-gallate (EGCG) is widely used as a weight controlling supplement. Concerns about its safety evoked after cases of hepatotoxicity occurred upon its use. The underlying factors that could be involved in EGCG associated hepatotoxicity are not fully studied. In this study, we investigated the possible impact of lipopolysaccharide (LPS), as an inflammagen, on the effect of EGCG on hepatocytes. HepG2 cells were treated with different concentrations of EGCG (100, 200, 500 μM), with and without LPS (10 nM)-presensitization of the cells. Viability of HepG2 cells decreased with the increased concentrations of EGCG; the viability was even lesser in LPS-presensitized cells. Oxidative stress (Ox.LDL and CXCL16), the expression of nuclear retinoic receptors (RAR, RXR) and the biomarkers of hepatocellular injury (TNFα, TGFβ1) were all relatively higher in LPS-presensitized cells compared to non-sensitized cells upon treatment with EGCG. Sensitization of HepG2 cells with LPS alone did not affect the viability or any of the other biomarkers considered in this study. In conclusion, EGCG alone can be harmful to liver at high concentrations and this effect may become more pronounced under the influence of an inflammagen.
- Keywords
- epigallocatechin-3-gallate (EGCG),
- MeSH
- Hep G2 Cells chemistry immunology drug effects MeSH
- Hepatocytes * chemistry immunology drug effects MeSH
- Catechin * analogs & derivatives pharmacology toxicity MeSH
- Polyphenols pharmacology toxicity MeSH
- In Vitro Techniques methods statistics & numerical data MeSH
- Cell Survival drug effects MeSH
- Inflammation * MeSH
- Publication type
- Evaluation Study MeSH
- Research Support, Non-U.S. Gov't MeSH
