Disruption of the thyroid hormone (TH) system is connected with diverse adverse health outcomes in wildlife and humans. It is crucial to develop and validate suitable in vitro assays capable of measuring the disruption of the thyroid hormone (TH) system. These assays are also essential to comply with the 3R principles, aiming to replace the ex vivo tests often utilised in the chemical assessment. We compared the two commonly used assays applicable for high throughput screening [Luminol and Amplex UltraRed (AUR)] for the assessment of inhibition of thyroid peroxidase (TPO, a crucial enzyme in TH synthesis) using several cell lines and 21 compounds from different use categories. As the investigated cell lines derived from human and rat thyroid showed low or undetectable TPO expression, we developed a series of novel cell lines overexpressing human TPO protein. The HEK-TPOA7 model was prioritised for further research based on the high and stable TPO gene and protein expression. Notably, the Luminol assay detected significant peroxidase activity and signal inhibition even in Nthy-ori 3-1 and HEK293T cell lines without TPO expression, revealing its lack of specificity. Conversely, the AUR assay was specific to TPO activity. Nevertheless, despite the different specificity, both assays identified similar peroxidation inhibitors. Over half of the tested chemicals with diverse structures and from different use groups caused TPO inhibition, including some widespread environmental contaminants suggesting a potential impact of environmental chemicals on TH synthesis. Furthermore, in silico SeqAPASS analysis confirmed the high similarity of human TPO across mammals and other vertebrate classes, suggesting the applicability of HEK-TPOA7 model findings to other vertebrates.
- MeSH
- autoantigeny metabolismus MeSH
- buněčné linie MeSH
- endokrinní disruptory toxicita MeSH
- HEK293 buňky MeSH
- jodidperoxidasa * antagonisté a inhibitory metabolismus genetika MeSH
- krysa rodu rattus MeSH
- lidé MeSH
- luminol MeSH
- oxaziny MeSH
- proteiny vázající železo metabolismus MeSH
- rychlé screeningové testy metody MeSH
- štítná žláza účinky léků metabolismus MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- srovnávací studie MeSH
Information on the indoor environment as a source of exposure with potential adverse health effects is mostly limited to a few pollutant groups and indoor types. This study provides a comprehensive toxicological profile of chemical mixtures associated with dust from various types of indoor environments, namely cars, houses, prefabricated apartments, kindergartens, offices, public spaces, and schools. Organic extracts of two different polarities and bioaccessible extracts mimicking the gastrointestinal conditions were prepared from two different particle size fractions of dust. These extracts were tested on a battery of human cell-based bioassays to assess endocrine disrupting potentials. Furthermore, 155 chemicals from different pollutant groups were measured and their relevance for the bioactivity was determined using concentration addition modelling. The exhaustive and bioaccessible extracts of dust from the different microenvironments interfered with aryl hydrocarbon receptor, estrogen, androgen, glucocorticoid, and thyroid hormone (TH) receptor signalling, and with TH transport. Noteably, bioaccessible extracts from offices and public spaces showed higher estrogenic effects than the organic solvent extracts. 114 of the 155 targeted chemicals were detectable, but the observed bioactivity could be only marginally explained by the detected chemicals. Diverse toxicity patterns across different microenvironments that people inhabit throughout their lifetime indicate potential health and developmental risks, especially for children. Limited data on the endocrine disrupting potency of relevant chemical classes, especially those deployed as replacements for legacy contaminants, requires further study.
Mikroskopická polyangiitida je ANCA vaskulitida, která se vyznačuje přítomností protilátek proti myeloperoxidáze (anti-MPO). Jedná se o vaskulitidu malých cév s fokálně nekrotizující glomerulonefritidou bez tvorby granulomů v dýchacích cestách. Na začátku onemocnění jsou přítomny nespecifické příznaky. Mezi ně patří artralgie, artritidy a myalgie. Difuzní alveolární hemoragie (DAH) je klinický syndrom charakterizovaný akutním nástupem alveolárních infiltrátů a hypoxemií, které vedou k progresivnímu difuznímu alveolárnímu krvácení vyžadujícího okamžitou léčbu. Kazuistika popisuje pacientku s nově diagnostikovanou mikroskopickou polyangiitidou (MPA), jejíž první manifestací byla difuzní alveolární hemoragie. Základní onemocnění bylo dále komplikováno těžkým hypoxickým respiračním selháním s nutností zavedení extrakorporální membránové oxygenace (V-V ECMO), obtížným weaningem (odpojení od umělé plicní ventilace) a recidivujícími infekty včetně multirezistentních kmenů. Pacientka byla léčena vysokodávkovými pulzy glukokortikoidů, cyklo- fosfamidem v redukované dávce a aplikací imunoglobulinů. Po osmiměsíční úspěšné léčbě byla propuštěna domů, nyní je její stav nadále uspokojivý.
Microscopic polyangiitis (MPA) is an ANCA-associated vasculitis characterized by the presence of anti-myeloperoxidase (anti-MPO) antibodies. It is a small vessel vasculitis with focally necrotizing glomerulonephritis without granuloma formation in the airways. Constitutional symptoms are present at the onset of the disease. These include arthralgia, arthritis, and myalgia. Diffuse alveolar hemorrhage (DAH) is a clinical syndrome characterized by acute onset of alveolar infiltrates and hypoxemia leading to progressive diffuse alveolar hemorrhage requiring immediate treatment. This case report describes a patient with newly diagnosed microscopic polyangiitis whose first manifestation was diffuse alveolar hemorrhage. The underlying disease was further complicated by severe hypoxic respiratory failure with the need for extracorporeal membrane oxygenation (V-V ECMO), difficult weaning (disconnection from artificial pulmonary ventilation), and recurrent infections including multidrug-resistant strains. The patient was treated with high-dose pulses of glucocorticoids, reduced-dose cyclophosphamide, and immunoglobulin administration. After eight months of successful treatment, she was discharged home and her condition remains satisfactory.
- Klíčová slova
- skin prick test,
- MeSH
- antikoagulancia * farmakologie terapeutické užití MeSH
- heparin nízkomolekulární farmakologie škodlivé účinky MeSH
- intradermální testy MeSH
- léková alergie * komplikace MeSH
- lidé MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- kazuistiky MeSH
Gaussian and exponentially modified Gaussian functions were incorporated into integrating algorithms used by an open-source, cross-platform tool called CycloBranch. The quantitation is demonstrated on bacterial pyoverdines separated by fine isotope features. Using our algorithm, we can separate the m/z values 694.25802 and 694.26731 (a 0.009 Da difference), where the former belongs to the most intense peak of pyoverdine D (PvdD), and the latter to the second most intense peak of pyoverdine E (PvdE) in the respective isotopic clusters of [M + Fe-H]2+ ions. The areas under chromatographic curves of standards were analyzed for the limit of detection (LOD), limit of quantitation (LOQ), and regression coefficient calculations. The quantitative module returned a LOD and LOQ of 1.4 and 4.3 ng/mL, respectively, for both PvdD and PvdE in human urine. If present and detected in mass spectra, the intensities of user-defined [M + H]+, [M + Na]+, [M + K]+, [M + Fe-H]2+, or other ion types, can be accumulated and used for quantitation. The quantitation result is returned by CycloBranch in seconds or minutes, contrary to an hours-long manual approach, prone to user-born errors originating from necessary copying among various software environments. Native Bruker, Waters, Thermo, txt, mgf, mzML, and mzXML data formats are supported in CycloBranch, which is freely available at https://ms.biomed.cas.cz/cyclobranch.
- MeSH
- algoritmy * MeSH
- chromatografie kapalinová metody MeSH
- hmotnostní spektrometrie metody MeSH
- izotopy MeSH
- lidé MeSH
- software * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- MeSH
- imunologické techniky metody MeSH
- klinické laboratorní techniky metody MeSH
- lidé středního věku MeSH
- lidé MeSH
- rektální píštěl MeSH
- Streptococcus oralis izolace a purifikace MeSH
- streptokokové infekce * diagnóza terapie MeSH
- únava etiologie MeSH
- výsledek terapie MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- kazuistiky MeSH
- MeSH
- familiární středomořská horečka diagnóza MeSH
- imunologické testy metody MeSH
- lidé středního věku MeSH
- lidé MeSH
- nežádoucí účinky léčiv diagnóza MeSH
- perikarditida diagnóza MeSH
- recidiva MeSH
- vakcína firmy Moderna proti COVID-19 škodlivé účinky MeSH
- zvýšená infektivita v přítomnosti protilátek * MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- kazuistiky MeSH
- MeSH
- běžná variabilní imunodeficience komplikace MeSH
- genetické nemoci vrozené MeSH
- intersticiální plicní nemoci * diagnóza etiologie terapie MeSH
- lidé středního věku MeSH
- lidé MeSH
- syndromy imunologické nedostatečnosti * genetika komplikace MeSH
- výsledek terapie MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- kazuistiky MeSH
