Chlorinated paraffins (CPs) are environmental pollutants extensively used in industries. While the use of short-chain chlorinated paraffins (SCCPs) has been restricted since 2017, the use of medium-chain chlorinated paraffins (MCCPs) has risen as their replacement. Due to lipophilic character, it can be expected that CPs enter the cells; however, the in vitro accumulation potential of CPs remains poorly understood. In this study, we aimed to explore the ability of SCCPs and MCCPs to accumulate in fat cells. We utilized an in vitro model of mouse 3T3-L1 preadipocytes and adipocytes. Using gas chromatography coupled with high-resolution mass spectrometry operated in negative chemical ionization mode, we determined the intracellular amounts of CPs. These compounds accumulated at rates of 8.5 ± 0.1 μg/gcells/h for SCCPs and 7.8 ± 0.3 μg/gcells/h for MCCPs when an initial concentration of 120 ng/ml was present in the medium. This rate increased approximately tenfold when the concentration of CPs was raised to 1200 ng/ml. CPs content in adipocytes steadily increased over 5 days, whereas preadipocytes accumulated 15-20 times less CPs. This highlights the importance of cellular lipid content, which was about 12 times higher in adipocytes. Furthermore, we found that the level of chlorine content in the CPs molecules significantly influenced their accumulation. Our results demonstrate that MCCPs exhibit a similar accumulation potential to SCCPs, with lipid content playing a crucial role. As with SCCPs, restrictions on the use of MCCPs in industry should be considered to mitigate their environmental and health impacts.
- MeSH
- buňky 3T3-L1 * MeSH
- chlorované uhlovodíky * metabolismus toxicita MeSH
- halogenace * MeSH
- látky znečišťující životní prostředí toxicita metabolismus MeSH
- metabolismus lipidů účinky léků MeSH
- myši MeSH
- parafín * MeSH
- plynová chromatografie s hmotnostně spektrometrickou detekcí MeSH
- tukové buňky * metabolismus účinky léků MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
In RNA interference (RNAi), long double-stranded RNA is cleaved by the Dicer endonuclease into small interfering RNAs (siRNAs), which guide degradation of complementary RNAs. While RNAi mediates antiviral innate immunity in plants and many invertebrates, vertebrates have adopted a sequence-independent response and their Dicer produces siRNAs inefficiently because it is adapted to process small hairpin microRNA precursors in the gene-regulating microRNA pathway. Mammalian endogenous RNAi is thus a rudimentary pathway of unclear significance. To investigate its antiviral potential, we modified the mouse Dicer locus to express a truncated variant (DicerΔHEL1) known to stimulate RNAi and we analyzed how DicerΔHEL1/wt mice respond to four RNA viruses: coxsackievirus B3 and encephalomyocarditis virus from Picornaviridae; tick-borne encephalitis virus from Flaviviridae; and lymphocytic choriomeningitis virus (LCMV) from Arenaviridae. Increased Dicer activity in DicerΔHEL1/wt mice did not elicit any antiviral effect, supporting an insignificant antiviral function of endogenous mammalian RNAi in vivo. However, we also observed that sufficiently high expression of DicerΔHEL1 suppressed LCMV in embryonic stem cells and in a transgenic mouse model. Altogether, mice with increased Dicer activity offer a new benchmark for identifying and studying viruses susceptible to mammalian RNAi in vivo.
- MeSH
- DEAD-box RNA-helikasy genetika metabolismus MeSH
- malá interferující RNA genetika MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- přirozená imunita * genetika MeSH
- ribonukleasa III * genetika metabolismus MeSH
- RNA interference * MeSH
- virus encefalomyokarditidy genetika imunologie MeSH
- virus lymfocytární choriomeningitidy imunologie genetika MeSH
- viry klíšťové encefalitidy genetika imunologie MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
PIWI-interacting RNAs (piRNAs) play a crucial role in safeguarding genome integrity by silencing mobile genetic elements. From flies to humans, piRNAs originate from long single-stranded precursors encoded by genomic piRNA clusters. How piRNA clusters form to adapt to genomic invaders and evolve to maintain protection remain key outstanding questions. Here, we generate a roadmap of piRNA clusters across seven species that highlights both similarities and variations. In mammals, we identify transcriptional readthrough as a mechanism to generate piRNAs from transposon insertions (piCs) downstream of genes (DoG). Together with the well-known stress-dependent DoG transcripts, our findings suggest a molecular mechanism for the formation of piRNA clusters in response to retroviral invasion. Finally, we identify a class of dynamic piRNA clusters in humans, underscoring unique features of human germ cell biology. Our results advance the understanding of conserved principles and species-specific variations in piRNA biology and provide tools for future studies.
- MeSH
- druhová specificita MeSH
- lidé MeSH
- malá interferující RNA * metabolismus genetika MeSH
- myši MeSH
- Piwi-interagující RNA MeSH
- psi MeSH
- savci * genetika MeSH
- transpozibilní elementy DNA genetika MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- psi MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- srovnávací studie MeSH
BACKGROUND: Sodium-glucose cotransporter 2 inhibitors (SGLT-2i) are glucose-lowering agents used for the treatment of type 2 diabetes mellitus, which also improve heart failure and decrease the risk of cardiovascular complications. Epicardial adipose tissue (EAT) dysfunction was suggested to contribute to the development of heart failure. We aimed to elucidate a possible role of changes in EAT metabolic and inflammatory profile in the beneficial cardioprotective effects of SGLT-2i in subjects with severe heart failure. METHODS: 26 subjects with severe heart failure, with reduced ejection fraction, treated with SGLT-2i versus 26 subjects without treatment, matched for age (54.0 ± 2.1 vs. 55.3 ± 2.1 years, n.s.), body mass index (27.8 ± 0.9 vs. 28.8 ± 1.0 kg/m2, n.s.) and left ventricular ejection fraction (20.7 ± 0.5 vs. 23.2 ± 1.7%, n.s.), who were scheduled for heart transplantation or mechanical support implantation, were included in the study. A complex metabolomic and gene expression analysis of EAT obtained during surgery was performed. RESULTS: SGLT-2i ameliorated inflammation, as evidenced by the improved gene expression profile of pro-inflammatory genes in adipose tissue and decreased infiltration of immune cells into EAT. Enrichment of ether lipids with oleic acid noted on metabolomic analysis suggests a reduced disposition to ferroptosis, potentially further contributing to decreased oxidative stress in EAT of SGLT-2i treated subjects. CONCLUSIONS: Our results show decreased inflammation in EAT of patients with severe heart failure treated by SGLT-2i, as compared to patients with heart failure without this therapy. Modulation of EAT inflammatory and metabolic status could represent a novel mechanism behind SGLT-2i-associated cardioprotective effects in patients with heart failure.
- MeSH
- antiflogistika terapeutické užití farmakologie MeSH
- biologické markery krev MeSH
- diabetes mellitus 2. typu farmakoterapie metabolismus diagnóza MeSH
- epikardiální adipózní tkáň MeSH
- funkce levé komory srdeční účinky léků MeSH
- glifloziny * terapeutické užití farmakologie škodlivé účinky MeSH
- lidé středního věku MeSH
- lidé MeSH
- mediátory zánětu * metabolismus MeSH
- metabolomika MeSH
- perikard * metabolismus účinky léků MeSH
- srdeční selhání * metabolismus patofyziologie farmakoterapie MeSH
- stupeň závažnosti nemoci * MeSH
- tepový objem účinky léků MeSH
- tuková tkáň * účinky léků metabolismus MeSH
- výsledek terapie MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
Canonical RNA interference (RNAi) is sequence-specific mRNA degradation guided by small interfering RNAs (siRNAs) made by RNase III Dicer from long double-stranded RNA (dsRNA). RNAi roles include gene regulation, antiviral immunity or defense against transposable elements. In mammals, RNAi is constrained by Dicer's adaptation to produce another small RNA class-microRNAs. However, a truncated Dicer isoform (ΔHEL1) supporting RNAi exists in mouse oocytes. A homozygous mutation to express only the truncated ΔHEL1 variant causes dysregulation of microRNAs and perinatal lethality in mice. Here, we report the phenotype and canonical RNAi activity in DicerΔHEL1/wt mice, which are viable, show minimal miRNome changes, but their endogenous siRNA levels are an order of magnitude higher. We show that siRNA production in vivo is limited by available dsRNA, but not by Protein kinase R, a dsRNA sensor of innate immunity. dsRNA expression from a transgene yields sufficient siRNA levels to induce efficient RNAi in heart and muscle. DicerΔHEL1/wt mice with enhanced canonical RNAi offer a platform for examining potential and limits of mammalian RNAi in vivo.
- MeSH
- DEAD-box RNA-helikasy genetika metabolismus MeSH
- dvouvláknová RNA * metabolismus genetika MeSH
- malá interferující RNA * genetika metabolismus MeSH
- mikro RNA genetika metabolismus MeSH
- myši MeSH
- protein - isoformy genetika metabolismus MeSH
- ribonukleasa III * genetika metabolismus MeSH
- RNA interference * MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Úvod: Ve své klasické podobě je onemocnění covid-19 respirační chorobou, která však může mít značně heterogenní průběh. Hrudní komplikace vznikají u malého procenta nemocných. Naším cílem je vyhodnotit dosavadní zkušenosti s touto chorobou a jejími hrudními projevy a stanovení reálného stavu péče o tyto nemocné. Metody: Tato práce je retrospektivní, single-insitution analýzou souboru pacientů hospitalizovaných s akutní a postakutní covid-19 pneumonií v pražské Thomayerově nemocnici v období 12/2020 až 03/2022, u kterých byl indikován hrudně-chirurgický zákrok. Výsledky: V době vrcholící pandemie nemoci covid-19 jsme provedli hrudně chirurgický zákrok u 46 pacientů. Nejčastější torakochirurgickou intervencí byla hrudní drenáž (18× pro pneumotorax, 3× pro fluidotorax, 2× CT navigovaná drenáž plicního abscesu a 2× CT navigovaná drenáž pneumatokély). V 10 případech byla provedena pleurektomie s dekortikací. Dále bylo provedeno 12 plicní parenchym šetřících resekcí, přičemž ve dvou případech si stav vyžádal resekci celého laloku. U 2 pacientů byla indikována resekce postintubační stenózy trachey po těžkém průběhu pneumonie covid-19. Závěr: Onemocnění covid-19 může i po lehkém průběhu výrazně morfologicky a funkčně alterovat dýchací ústrojí. Nejčastějšími komplikacemi pneumonie covid-19 vyžadující torakochirurgickou intervenci jsou patologie spojené se vznikem airleaku s kumulací vzduchu (pneumotorax, pneumomediastinum a podkožní emfyzém). Rozvoj plicních nekróz, symptomatických bronchiektázií, pneumatokél, bulózně-fibrotických formací mohou v ojedinělých případech vyústit v pneumotorax, hemotorax nebo hrudní empyém. Včasná torakochirurgická intervence hrudních komplikací pneumonie covid-19 může vést ke zlepšení přežívání covid nemocných.
Introduction: COVID-19 is considered a respiratory virosis in its classic form, although it may present with heterogeneous symptoms. Thoracic complications occur in a small percentage of patients. Our objective was to evaluate existing experience with this disease and its thoracic manifestations and to determine the real-world status of care of these patients. Methods: This study is a retrospective, single-institution analysis of a group of patients hospitalized with acute and post-acute COVID-19 pneumonia at Thomayer Hospital in Prague in the period from December 2020 to March 2022 and indicated for a thoracic surgical procedure. Results: During the peak of COVID-19 pandemic, a thoracic intervention was performed in 46 admitted patients. Thoracic drainage (due to pneumothorax in 18 cases, fluidothorax in 3 cases, CT-guided lung abscess drainage in 2 cases, and CT-guided pneumatocele drainage in 2 cases) were the most common thoracic surgical procedures. Pleurectomy/decortication surgery was done in 10 cases. Additionally, 12 lung parenchyma-sparing resections were performed, while lobectomy was required in 2 cases. Resection of postintubation tracheal stenosis due to a severe course of COVID-19 pneumonia was indicated in 2 patients. Conclusion: Even mild COVID-19 may cause a considerable morphological a functional alteration of the respiratory system. The most common complications of COVID-19 pneumonia that require a thoracic surgical intervention include pathologies associated with an air leak and accumulation of air (pneumothorax, pneumomediastinum and subcutaneous emphysema). The development of pulmonary necrosis, symptomatic bronchiectasis, pneumatocele, and bullous-fibrotic formations may result in pneumothorax, hemothorax or thoracic empyema in sporadic cases. An early thoracic surgical intervention to treat thoracic complications of COVID-19 pneumonia can improve the survival of COVID-19 patients.
- MeSH
- chirurgie plic metody statistika a číselné údaje MeSH
- hrudní chirurgické výkony * metody statistika a číselné údaje MeSH
- lidé středního věku MeSH
- lidé MeSH
- pneumonie chirurgie etiologie komplikace patologie MeSH
- počítačová rentgenová tomografie MeSH
- postakutní syndrom COVID-19 * chirurgie diagnostické zobrazování mortalita patologie MeSH
- senioři MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- klinická studie MeSH
Endogenous opioid peptides serve as potent analgesics through the opioid receptor (OR) activation. However, they often suffer from poor metabolic stability, low lipophilicity, and low blood-brain barrier permeability. Researchers have developed many strategies to overcome the drawbacks of current pain medications and unwanted biological effects produced by the interaction with opioid receptors. Here, we tested multifunctional enkephalin analogs LYS739 (MOR/DOR agonist and KOR partial antagonist) and LYS744 (MOR/DOR agonist and KOR full antagonist) under in vivo conditions in comparison with MOR agonist, morphine. We applied 2D electrophoretic resolution to investigate differences in proteome profiles of crude membrane (CM) fractions isolated from the rat brain cortex and hippocampus exposed to the drugs (10 mg/kg, seven days). Our results have shown that treatment with analog LYS739 induced the most protein changes in cortical and hippocampal samples. The identified proteins were mainly associated with energy metabolism, cell shape and movement, apoptosis, protein folding, regulation of redox homeostasis, and signal transduction. Among these, the isoform of mitochondrial ATP synthase subunit beta (ATP5F1B) was the only protein upregulation in the hippocampus but not in the brain cortex. Contrarily, the administration of analog LYS744 caused a small number of protein alterations in both brain parts. Our results indicate that the KOR full antagonism, together with MOR/DOR agonism of multifunctional opioid ligands, can be beneficial in treating chronic pain states by reducing changes in protein expression levels but retaining analgesic efficacy.
- MeSH
- analgetika MeSH
- enkefaliny metabolismus MeSH
- hipokampus metabolismus MeSH
- krysa rodu rattus MeSH
- morfin * farmakologie MeSH
- mozek metabolismus MeSH
- opioidní analgetika farmakologie MeSH
- receptory opiátové mu * metabolismus MeSH
- receptory opiátové metabolismus MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
