This study elucidated the impacts of coenzyme Q10 (COQ10) supplementation in a high-fat diet (HFD) on growth, lipid metabolism and mitochondrial function in spotted seabass (Lateolabrax maculatus). Totally five diets were formulated: a diet with normal fat content (11 % lipid, NFD), a HFD (17 % lipid) and three additional diets by supplementing 5, 20 or 80 mg/kg of COQ10 to the HFD. After an 8-week culture period, samples were collected and analysed. The results demonstrated that COQ10 inclusion prevented the HFD-induced deterioration of growth performance and feed utilisation. COQ10 alleviated the deposition of saturated fatty acids following HFD intake and promoted the assimilation of n-3 and n-6 PUFA. Moreover, COQ10 administration inhibited the surge in serum transaminase activity and reduced hepatic lipid content following HFD ingestion, which was consistent with the results of oil red O staining. In addition, HFD feeding led to reduced hepatic citrate synthase and succinate dehydrogenase activities and decreased ATP content. Notably, COQ10 administration improved these indices and up-regulated the expression of mitochondrial biogenesis-related genes (pgc-1α, pgc-1β, nrf-1, tfam) and autophagy-related genes (pink1, mul1, atg5). In summary, supplementing 20-80 mg/kg of COQ10 in the HFD promoted growth performance, alleviated hepatic fat accumulation and enhanced liver mitochondrial function in spotted seabass.

• MeSH
• Diet, High-Fat * adverse effects   MeSH
• Liver metabolism   drug effects   MeSH
• Animal Feed analysis   MeSH
• Lipid Metabolism drug effects   MeSH
• Mitochondria * drug effects   metabolism   MeSH
• Bass * growth & development   metabolism   MeSH
• Dietary Supplements MeSH
• Ubiquinone * analogs & derivatives   pharmacology   administration & dosage   MeSH
• Fatty Liver * veterinary   etiology   drug therapy   prevention & control   MeSH
• Animals MeSH
• Check Tag
• Animals MeSH
• Publication type
• Journal Article MeSH

UNLABELLED: The paper presents the study of a set of isolates of Streptococcus pneumoniae, which comprised two heterogeneous subpopulations, one of which was susceptible and the other resistant to optochin. The aim of the study was to compare the results of serotyping, multilocus sequence typing (MLST), ribosomal multilocus sequence typing (rMLST), and variation analysis of these subpopulations and to investigate the genetic probable causes of optochin resistance. The strains studied were cultured from samples taken from patients with invasive pneumococcal disease in the Czech Republic in 2019 and 2020. A total of 10 studied pairs of isolates were subject to serotyping and whole-genome sequencing (WGS). None of the typing methods (serotyping, MLST, or rMLST) applied to pairs of optochin-susceptible and optochin-resistant isolates revealed differences in serotype, sequence type, or ribosomal sequence type. The WGS data analysis identified point mutations in ATP (adenosine triphosphate) synthase genes in 8 of the 10 optochin-resistant isolates. In seven optochin-resistant isolates, the mutation was found in the atpC gene and in one isolate in the atpA gene. One of the mutations in the atpC gene has not yet been published in the literature; it is a mutation at position 143T > C with an amino acid change of Val48Ala. In 8 out of the 10 optochin-resistant isolates, the possible genetic basis for resistance was identified, involving point mutations in the atpA and atpC genes. In the remaining two isolates, no clear genetic explanation for the optochin resistance in S. pneumoniae was found, based on current knowledge. IMPORTANCE: Globally, among the most fundamental tests used for the identification of Streptococcus pneumoniae isolates is determining susceptibility to optochin. In the last 2 decades, optochin-resistant strains have been frequently reported in the literature, which can lead to the misidentification of S. pneumoniae. This study compares whole-genome sequencing data of optochin-susceptible and optochin-resistant subpopulations of S. pneumoniae isolates and investigates the genetic probable causes of resistance in the genomes of optochin-resistant subpopulations.

• MeSH
• Anti-Bacterial Agents * pharmacology   MeSH
• Drug Resistance, Bacterial * genetics   MeSH
• Bacterial Proteins genetics   MeSH
• Quinine analogs & derivatives   MeSH
• Genome, Bacterial MeSH
• Humans MeSH
• Microbial Sensitivity Tests MeSH
• Multilocus Sequence Typing MeSH
• Pneumococcal Infections microbiology   MeSH
• Whole Genome Sequencing MeSH
• Serotyping MeSH
• Streptococcus pneumoniae * genetics   drug effects   isolation & purification   classification   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Comparative Study MeSH
• Geographicals
• Czech Republic MeSH

DNA ligase 1 (LIG1) plays a key role in DNA synthesis and DNA damage repair pathways. LIG1 has been shown to be up-regulated in human non-small cell lung cancer (NSCLC); however, its role and molecular regulatory mechanism in NSCLC cell proliferation are still not fully understand. In this study, we aimed to explore the role of LIG1 and post-transcripional regulators in NSCLC. Utilizing bioinformatic tools and qRT-PCR, our investigation substantiated the up-regulation of LIG1 within NSCLC cell lines and tumour tissues. Remarkably, individuals exhibiting elevated levels of LIG1 had diminished survival rates. Functionally, the depletion of LIG1 inhibited cell proliferation and migration, contrasting with the increased proliferation and migration upon LIG1 over-expression. Prediction from the TargetScanHuman database and results of dual luciferase reporter assays indicated that miR-325 could directly bind to and negatively regulate LIG1. Moreover, our findings demonstrated that the mimicry of miR-325 decreased cell viability, whereas its inhibition correspondingly increased viability, indicative of the tumour-suppressive role of miR-325 through the down-regulation of LIG1. Collectively, our findings show that LIG1 could promote tumour progression and knockdown of LIG1 could exert suppressive effects on NSCLC. As the post-transcriptional factor of LIG1, miR-325 could negatively regulate the expression of LIG1 to inhibit tumour progression in vitro. These findings suggest that LIG1 and miR-325 might be potential therapeutic targets for NSCLC treatment.

• MeSH
• DNA Ligase ATP * metabolism   genetics   MeSH
• Humans MeSH
• MicroRNAs * genetics   metabolism   MeSH
• Cell Line, Tumor MeSH
• Lung Neoplasms * pathology   genetics   metabolism   MeSH
• Carcinoma, Non-Small-Cell Lung * genetics   pathology   metabolism   MeSH
• Cell Movement * MeSH
• Cell Proliferation * MeSH
• Gene Expression Regulation, Neoplastic MeSH
• Check Tag
• Humans MeSH
• Male MeSH
• Female MeSH
• Publication type
• Journal Article MeSH

Replication forks stalled at co-transcriptional R-loops can be restarted by a mechanism involving fork cleavage-religation cycles mediated by MUS81 endonuclease and DNA ligase IV (LIG4), which presumably relieve the topological barrier generated by the transcription-replication conflict (TRC) and facilitate ELL-dependent reactivation of transcription. Here, we report that the restart of R-loop-stalled replication forks via the MUS81-LIG4-ELL pathway requires senataxin (SETX), a helicase that can unwind RNA:DNA hybrids. We found that SETX promotes replication fork progression by preventing R-loop accumulation during S-phase. Interestingly, loss of SETX helicase activity leads to nascent DNA degradation upon induction of R-loop-mediated fork stalling by hydroxyurea. This fork degradation phenotype is independent of replication fork reversal and results from DNA2-mediated resection of MUS81-cleaved replication forks that accumulate due to defective replication restart. Finally, we demonstrate that SETX acts in a common pathway with the DEAD-box helicase DDX17 to suppress R-loop-mediated replication stress in human cells. A possible cooperation between these RNA/DNA helicases in R-loop unwinding at TRC sites is discussed.

• MeSH
• Flap Endonucleases metabolism   genetics   MeSH
• DEAD-box RNA Helicases * metabolism   genetics   MeSH
• DNA-Binding Proteins * metabolism   genetics   MeSH
• DNA Helicases * metabolism   genetics   MeSH
• DNA Ligase ATP metabolism   genetics   MeSH
• DNA metabolism   genetics   MeSH
• Endonucleases * metabolism   genetics   MeSH
• Transcription, Genetic MeSH
• Humans MeSH
• Multifunctional Enzymes * metabolism   genetics   MeSH
• R-Loop Structures * MeSH
• DNA Replication * MeSH
• RNA Helicases * metabolism   genetics   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH

• MeSH
• Biochemistry MeSH
• Metabolic Diseases MeSH
• Pathology MeSH

• Conspectus
• Patologie. Klinická medicína
• Učební osnovy. Vyučovací předměty. Učebnice
• NML Fields
• biochemie
• patologie
• NML Publication type
• učebnice vysokých škol

Hyperhomocysteinemia (HHcy) is considered an independent risk factor of cardiovascular diseases. Among the proposed mechanisms underlying homocysteine toxicity are altered protein expression and induction of oxidative stress. In the present study, we explored protein abundance and parameters related to oxidative stress in heart homogenates of rats exposed to chronic mild HHcy. Using two-dimensional gel electrophoresis followed by MALDI-TOF/TOF mass spectrometry 22 altered proteins (6 upregulated and 14 downregulated) were identified. For eight proteins the altered abundances were validated by Western blot analysis. Identified proteins are primarily involved in energy metabolism (mainly enzymes of glycolysis, pyruvate dehydrogenase complex, citric acid cycle, and ATP synthase), cardiac muscle contraction (alpha-actin and myosin light chains), stress response (heat-shock protein beta1 and alphaB-crystallin) and antioxidant defense (glutathione peroxidase 1). Diminished antioxidant defense was confirmed by decreases in total antioxidant capacity and GSH/GSSG ratio. Consistent with the decline in enzymatic and non-enzymatic antioxidant defense the protein oxidative modification, as determined by tyrosine nitration, was significantly increased. These findings suggest that both, altered protein expression and elevated oxidative stress contribute to cardiovascular injury caused by HHcy. Keywords: Homocysteine, Heart, Protein abundance, Antioxidant capacity, Nitrotyrosines.

• MeSH
• Hyperhomocysteinemia * metabolism   MeSH
• Rats MeSH
• Myocardium * metabolism   MeSH
• Oxidative Stress * MeSH
• Rats, Wistar * MeSH
• Animals MeSH
• Check Tag
• Rats MeSH
• Male MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

Mitochondrial oxidative phosphorylation (OXPHOS) fuels cellular ATP demands. OXPHOS defects lead to severe human disorders with unexplained tissue specific pathologies. Mitochondrial gene expression is essential for OXPHOS biogenesis since core subunits of the complexes are mitochondrial-encoded. COX14 is required for translation of COX1, the central mitochondrial-encoded subunit of complex IV. Here we describe a COX14 mutant mouse corresponding to a patient with complex IV deficiency. COX14M19I mice display broad tissue-specific pathologies. A hallmark phenotype is severe liver inflammation linked to release of mitochondrial RNA into the cytosol sensed by RIG-1 pathway. We find that mitochondrial RNA release is triggered by increased reactive oxygen species production in the deficiency of complex IV. Additionally, we describe a COA3Y72C mouse, affected in an assembly factor that cooperates with COX14 in early COX1 biogenesis, which displays a similar yet milder inflammatory phenotype. Our study provides insight into a link between defective mitochondrial gene expression and tissue-specific inflammation.

• MeSH
• Cyclooxygenase 1 * MeSH
• DEAD Box Protein 58 MeSH
• DEAD-box RNA Helicases metabolism   genetics   MeSH
• Liver * metabolism   pathology   MeSH
• Humans MeSH
• Membrane Proteins MeSH
• Mitochondrial Proteins metabolism   genetics   MeSH
• Mitochondria metabolism   MeSH
• Mutation MeSH
• Mice, Inbred C57BL MeSH
• Mice MeSH
• Oxidative Phosphorylation * MeSH
• Protein Biosynthesis MeSH
• Reactive Oxygen Species * metabolism   MeSH
• Electron Transport Complex IV * metabolism   genetics   MeSH
• RNA, Mitochondrial genetics   metabolism   MeSH
• Inflammation * metabolism   genetics   pathology   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Male MeSH
• Mice MeSH
• Female MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

Disorders of ATP synthase, the key enzyme in mitochondrial energy supply, belong to the most severe metabolic diseases, manifesting as early-onset mitochondrial encephalo-cardiomyopathies. Since ATP synthase subunits are encoded by both mitochondrial and nuclear DNA, pathogenic variants can be found in either genome. In addition, the biogenesis of ATP synthase requires several assembly factors, some of which are also hotspots for pathogenic variants. While variants of MT-ATP6 and TMEM70 represent the most common cases of mitochondrial and nuclear DNA mutations respectively, the advent of next-generation sequencing has revealed new pathogenic variants in a number of structural genes and TMEM70, sometimes with truly peculiar genetics. Here we present a systematic review of the reported cases and discuss biochemical mechanisms, through which they are affecting ATP synthase. We explore how the knowledge of pathophysiology can improve our understanding of enzyme biogenesis and function. Keywords: Mitochondrial diseases o ATP synthase o Nuclear DNA o Mitochondrial DNA o TMEM70.

• MeSH
• Phenotype * MeSH
• Humans MeSH
• Membrane Proteins genetics   metabolism   MeSH
• DNA, Mitochondrial genetics   MeSH
• Mitochondrial Diseases genetics   enzymology   MeSH
• Mitochondrial Proteins genetics   metabolism   MeSH
• Mitochondrial Proton-Translocating ATPases * genetics   metabolism   MeSH
• Mitochondria enzymology   genetics   MeSH
• Mutation MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Review MeSH
• Systematic Review 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
• Analgesics MeSH
• Enkephalins metabolism   MeSH
• Hippocampus metabolism   MeSH
• Rats MeSH
• Morphine * pharmacology   MeSH
• Brain metabolism   MeSH
• Analgesics, Opioid pharmacology   MeSH
• Receptors, Opioid, mu * metabolism   MeSH
• Receptors, Opioid metabolism   MeSH
• Animals MeSH
• Check Tag
• Rats MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

Mycobacterial ATP synthase is a validated therapeutic target for combating drug-resistant tuberculosis. Inhibition of this enzyme has been featured as an efficient strategy for the development of new antimycobacterial agents against drug-resistant pathogens. In this study, we synthesised and explored two distinct series of squaric acid analogues designed to inhibit mycobacterial ATP synthase. Among the extensive array of compounds investigated, members of the phenyl-substituted sub-library emerged as primary hits. To gain deeper insights into their mechanisms of action, we conducted advanced biological studies, focusing on the compounds displaying a direct binding of a nitrogen heteroatom to the phenyl ring, resulting in the highest potency. Our investigations into spontaneous mutants led to the validation of a single point mutation within the atpB gene (Rv1304), responsible for encoding the ATP synthase subunit a. This genetic alteration sheds light on the molecular basis of resistance to squaramides. Furthermore, we explored the possibility of synergy between squaramides and the reference drug clofazimine using a checkerboard assay, highlighting the promising avenue for enhancing the effectiveness of existing treatments through combined therapeutic approaches. This study contributes to the expansion of investigating squaramides as promising drug candidates in the ongoing battle against drug-resistant tuberculosis.

• MeSH
• Adenosine Triphosphate metabolism   MeSH
• Antitubercular Agents chemistry   MeSH
• Humans MeSH
• Mitochondrial Proton-Translocating ATPases chemistry   metabolism   MeSH
• Tuberculosis, Multidrug-Resistant * MeSH
• Mycobacterium tuberculosis * MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH