Alternative DNA structures play critical roles in fundamental biological processes linked to human diseases. Thus, targeting and stabilizing these structures by specific ligands could affect the progression of cancer and other diseases. Here, we describe, using methods of molecular biophysics, the interactions of two oxidatively locked [Co2L3]6+ cylinders, rac-2 and meso-1, with diverse alternative DNA structures, such as junctions, G quadruplexes, and bulges. This study was motivated by earlier results demonstrating that both Co(III) cylinders exhibit potent and selective activity against cancer cells, accumulate in the nucleus of cancer cells, and prove to be efficient DNA binders. The results show that the bigger cylinder rac-2 stabilizes all DNA structures, while the smaller cylinder meso-1 stabilizes just the Y-shaped three-way junctions. Collectively, the results of this study suggest that the stabilization of alternative DNA structures by Co(III) cylinders investigated in this work might contribute to the mechanism of their biological activity.

• Klíčová slova
• Alternative DNA structures, Cobalt helicates, DNA G-quadruplexes, DNA bulges, DNA junctions,
• MeSH
• DNA * chemie   metabolismus   MeSH
• G-kvadruplexy MeSH
• kobalt * chemie   MeSH
• komplexní sloučeniny chemie   farmakologie   MeSH
• konformace nukleové kyseliny MeSH
• lidé MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Nucleic acids are not only static carriers of genetic information but also play vital roles in controlling cellular lifecycles through their fascinating structural diversity [...].

• MeSH
• DNA * chemie   metabolismus   MeSH
• konformace nukleové kyseliny * MeSH
• lidé MeSH
• RNA * chemie   metabolismus   MeSH
• výpočetní biologie * metody   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• úvodní články MeSH
• úvodníky MeSH
• Názvy látek
• DNA * MeSH
• RNA * MeSH

A series of triterpenoid pyrones was synthesized and subsequently modified to introduce phthalimide or phthalate moieties into the triterpenoid skeleton. These compounds underwent in vitro cytotoxicity screening, revealing that a subset of six compounds exhibited potent activity, with IC50 values in the low micromolar range. Further biological evaluations, including Annexin V and propidium iodide staining experiment revealed, that all compounds induce selective apoptosis in cancer cells. Measurements of mitochondrial potential, cell cycle analysis, and the expression of pro- and anti-apoptotic proteins confirmed, that apoptosis was mediated via the mitochondrial pathway. These findings were further supported by cell cycle modulation and DNA/RNA synthesis studies, which indicated a significant increase in cell accumulation in the G0/G1 phase and a marked reduction in S-phase cells, alongside a substantial inhibition of DNA synthesis. The activation of caspase-3 and the cleavage of PARP, coupled with a decrease in the expression of Bcl-2 and Bcl-XL proteins, underscored the induction of apoptosis through the mitochondrial pathway. Given their high activity and pronounced effect on mitochondria function, trifluoromethyl pyrones 1f and 2f, and dihydrophthalimide 2h have been selected for further development.

• Klíčová slova
• Apoptosis, Cancer, Cytotoxicity, Heterocycle, Mitochondria, Pharmacology, Phthalate, Phthalimide, Pyrone, Triterpene,
• MeSH
• antitumorózní látky * terapeutické užití   MeSH
• apoptóza MeSH
• DNA metabolismus   MeSH
• ftalimidy farmakologie   MeSH
• kyseliny ftalové * MeSH
• membránový potenciál mitochondrií MeSH
• mitochondrie metabolismus   MeSH
• nádorové buněčné linie MeSH
• nádory * farmakoterapie   MeSH
• pyrony farmakologie   MeSH
• triterpeny * farmakologie   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antitumorózní látky * MeSH
• DNA MeSH
• ftalimidy MeSH
• kyseliny ftalové * MeSH
• phthalic acid MeSH Prohlížeč
• pyrony MeSH
• triterpeny * MeSH

Short-term sperm storage is a straightforward and cost-effective method of managing logistics in large scale fish hatchery operations but may result in decline in sperm quality. For effective artificial reproduction of fish, use of an appropriate additive to optimize sperm storage conditions is essential. In this study, it was investigated the effect of purified seminal plasma transferrin (Tf) at 10 μg/ml on relevant parameters in common carp Cyprinus carpio sperm during short-term storage. We compared sperm motility and curvilinear velocity, adenosine triphosphate (ATP) content and DNA fragmentation of fresh spermatozoa to that stored for 24, 48, 72, and 144 h with or without Tf. The percentage of motile cells and the curvilinear velocity of spermatozoa in stored samples for 72 h with transferrin supplementation were greater compared to samples with no added protein. The ATP content in samples without added transferrin was reduced (P < 0.05) after 72 h of storage, in contrast to the levels observed in transferrin-supplemented sperm. A time-dependent increase in DNA fragmentation was observed. Significantly lower DNA damage, expressed as percent tail DNA (10.99 ± 1.28) and olive tail moment (0.54 ± 0.12), was recorded in Tf-supplemented samples stored for 48 h compared to that with no Tf. Hence, it is concluded that the beneficial effects of transferrin on common carp sperm could serve as an additional tool for developing and enhancing short-term sperm preservation procedures commonly used in aquaculture.

• Klíčová slova
• Adenosine triphosphate, Cyprinidae, DNA fragmentation, Sperm motility, Sperm storage,
• MeSH
• adenosintrifosfát metabolismus   MeSH
• DNA metabolismus   MeSH
• kapři * MeSH
• motilita spermií MeSH
• sperma metabolismus   MeSH
• spermie MeSH
• transferin farmakologie   MeSH
• uchování spermatu * veterinární   metody   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• adenosintrifosfát MeSH
• DNA MeSH
• transferin MeSH

The objective of this study was to evaluate whether RSV inhibits neutrophil extracellular traps (NETs) that induce joint hyperalgesia in C57BL/6 mice after adjuvant-induced arthritis. A subplantar injection of Freund's complete adjuvant was administered to C57BL/6 mice on day 0 for immunization in the AIA model. Resveratrol (RSV, 25 mg/kg) was administered intraperitoneally once daily starting on day 22 and continuing for two weeks. The effects of mechanical hyperalgesia and edema formation have been assessed in addition to histopathological scoring. Mice were sacrificed on day 35 to determine cytokine levels and PADI4 and COX-2 expression levels. ELISA was used to quantify neutrophil extracellular traps (NETs) along with neutrophil elastase-DNA and myeloperoxidase-DNA complexes in neutrophils. An immunohistochemical stain was performed on knee joints to determine the presence of nuclear factor kappa B p65 (NF-kappaB p65). AIA mice were found to have higher levels of NET in joints and their joint cells demonstrated an increased expression of the PADI4 gene. Treatment with RSV in AIA mice (25 mg/kg, i.p.) significantly (P<0.05) inhibited joint hyperalgesia, resulting in a significant increase in mechanical threshold, a decrease in articular edema, a decrease in the production of inflammatory cytokines, increased COX-2 expression, and a decrease in the immunostaining of NF-kappaB. Furthermore, treatment with RSV significantly reduced the amount of neutrophil elastase (NE)-DNA and MPO-DNA complexes, which were used as indicators of NET formation (P<0.05). This study indicates that RSV reduces NET production and hyperalgesia by reducing inflammation mediated by PADI4 and COX-2. According to these data, NETs contribute to joint pain and resveratrol can be used to treat pain in RA through this pathway.

• MeSH
• cyklooxygenasa 2 MeSH
• cytokiny metabolismus   MeSH
• DNA metabolismus   MeSH
• edém metabolismus   MeSH
• extracelulární pasti * metabolismus   MeSH
• hyperalgezie farmakoterapie   metabolismus   MeSH
• leukocytární elastasa metabolismus   farmakologie   MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• neutrofily metabolismus   MeSH
• NF-kappa B metabolismus   MeSH
• resveratrol farmakologie   terapeutické užití   metabolismus   MeSH
• revmatoidní artritida * metabolismus   MeSH
• toll-like receptor 4 metabolismus   MeSH
• zánět metabolismus   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• cyklooxygenasa 2 MeSH
• cytokiny MeSH
• DNA MeSH
• leukocytární elastasa MeSH
• NF-kappa B MeSH
• resveratrol MeSH
• toll-like receptor 4 MeSH

Arsenic toxicity is a major problem across the world due to geogenic activity and has been supposed to generate free radicals and genotoxicity among the arsenic-poisoned population. There is a need to find suitable free radical quenching compounds for the arsenic-induced free radical-affected population. In the present study, Na3AsO3- induced oxidative stress and genotoxicity were evaluated in Oryctolagus cuniculus L, and quenching competency of Ocimum species was examined by applying enzymatic and non-enzymatic in vitro tests, comet assay, and Random Amplified Polymorphic Deoxyribonucleic acid - Polymerase Chain Reaction (RAPD-PCR) methods. In the present study, oxidative damage due to Na3AsO3 intoxication in O. cuniculus L has been confirmed followed by substantive genotoxicity, and in a further study, it has also been reported that the extract of O. gratissimum L lowers the oxidative stress in experimental animals confirmed by a decrease in Malondialdehyde (MDA) 4.78 ± 0.05 (nmol/mg protein), and an increase in Glutathione (GSH) 2.87 ± 0.50 (μmoles/mg proteins), Superoxide Dismutase (SOD) 1.78 ± 0.03(Units/mg protein), Catalase (CAT) 2.72 ± 0.02 (μmoles of H2O2 consumed/min/mg proteins) and Glutathione peroxidase (GPX) 7.43 ± 0.01 (μg of glutathione utilized/min/mg protein). A positive impact of extract of O. gratissimum L on protection of genotoxicity has been also confirmed by Random Amplified Polymorphic DNA (RAPD) based reduction in polymorphic bands of Deoxyribonucleic acid (DNA) from 6.5 to 3.16 and comet assay-based increase in head DNA % (87.86 ± 1.58), tail moment (1.07 ± 0.27) and decrease in tail DNA % (12.13 ± 1.58) & tail length (8.2 ± 1.46) at 5% P in lymphocytes. A significant level reduction in free radicals and reduction in DNA polymorphism has proved the competency of test material for the development of suitable antidotes against arsenicosis.

• Klíčová slova
• Arsenic, Comet assay, Lymphocytes, Oryctolagus cuniculus L., Oxidative stress, Phytochemicals,
• MeSH
• antioxidancia farmakologie   MeSH
• arsen * toxicita   MeSH
• bazalka * metabolismus   MeSH
• DNA metabolismus   MeSH
• glutathion metabolismus   MeSH
• králíci MeSH
• otrava arsenem * MeSH
• oxidační stres MeSH
• peroxid vodíku MeSH
• poškození DNA MeSH
• superoxiddismutasa metabolismus   MeSH
• technika náhodné amplifikace polymorfní DNA MeSH
• zvířata MeSH
• Check Tag
• králíci MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antioxidancia MeSH
• arsen * MeSH
• DNA MeSH
• glutathion MeSH
• peroxid vodíku MeSH
• superoxiddismutasa MeSH

The transcriptional co-activator lens epithelium-derived growth factor/p75 (LEDGF/p75) plays an important role in the biology of the cell and in several human diseases, including MLL-rearranged acute leukemia, autoimmunity, and HIV-1 infection. In both health and disease, LEDGF/p75 functions as a chromatin tether that interacts with proteins such as MLL1 and HIV-1 integrase via its integrase-binding domain (IBD) and with chromatin through its N-terminal PWWP domain. Recently, dimerization of LEDGF/p75 was shown, mediated by a network of electrostatic contacts between amino acids from the IBD and the C-terminal α6-helix. Here, we investigated the functional impact of LEDGF/p75 variants on the dimerization using biochemical and cellular interaction assays. The data demonstrate that the C-terminal α6-helix folds back in cis on the IBD of monomeric LEDGF/p75. We discovered that the presence of DNA stimulates LEDGF/p75 dimerization. LEDGF/p75 dimerization enhances binding to MLL1 but not to HIV-1 integrase, a finding that was observed in vitro and validated in cell culture. Whereas HIV-1 replication was not dependent on LEDGF/p75 dimerization, colony formation of MLLr-dependent human leukemic THP-1 cells was. In conclusion, our data indicate that intricate changes in the quaternary structure of LEDGF/p75 modulate its tethering function.

• Klíčová slova
• DNA-binding protein, DNA-induced protein binding, LEDGF/p75, chromatin structure, protein dynamic, protein–DNA interaction, protein–protein interaction,
• MeSH
• chromatin * MeSH
• dimerizace MeSH
• DNA metabolismus   MeSH
• lidé MeSH
• mezibuněčné signální peptidy a proteiny * metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• chromatin * MeSH
• DNA MeSH
• lens epithelium-derived growth factor MeSH Prohlížeč
• mezibuněčné signální peptidy a proteiny * MeSH

Non-canonical secondary structures in DNA are increasingly being revealed as critical players in DNA metabolism, including modulating the accessibility and activity of promoters. These structures comprise the so-called G-quadruplexes (G4s) that are formed from sequences rich in guanine bases. Using a well-defined transcriptional reporter system, we sought to systematically investigate the impact of the presence of G4 structures on transcription in yeast Saccharomyces cerevisiae. To this aim, different G4 prone sequences were modeled to vary the chance of intramolecular G4 formation, analyzed in vitro by Thioflavin T binding test and circular dichroism and then placed at the yeast ADE2 locus on chromosome XV, downstream and adjacent to a P53 response element (RE) and upstream from a minimal CYC1 promoter and Luciferase 1 (LUC1) reporter gene in isogenic strains. While the minimal CYC1 promoter provides basal reporter activity, the P53 RE enables LUC1 transactivation under the control of P53 family proteins expressed under the inducible GAL1 promoter. Thus, the impact of the different G4 prone sequences on both basal and P53 family protein-dependent expression was measured after shifting cells onto galactose containing medium. The results showed that the presence of G4 prone sequences upstream of a yeast minimal promoter increased its basal activity proportionally to their potential to form intramolecular G4 structures; consequently, this feature, when present near the target binding site of P53 family transcription factors, can be exploited to regulate the transcriptional activity of P53, P63 and P73 proteins.

• Klíčová slova
• G-quadruplex, p53, transcriptional activity, yeast,
• MeSH
• DNA metabolismus   MeSH
• G-kvadruplexy * MeSH
• nádorový supresorový protein p53 genetika   MeSH
• promotorové oblasti (genetika) MeSH
• Saccharomyces cerevisiae * genetika   metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• DNA MeSH
• nádorový supresorový protein p53 MeSH

The evolutionarily conserved DNA repair complex Ku serves as the primary sensor of free DNA ends in eukaryotic cells. Its rapid association with DNA ends is crucial for several cellular processes, including non-homologous end joining (NHEJ) DNA repair and telomere protection. In this study, we conducted a transient kinetic analysis to investigate the impact of the SAP domain on individual phases of the Ku-DNA interaction. Specifically, we examined the initial binding, the subsequent docking of Ku onto DNA, and sliding of Ku along DNA. Our findings revealed that the C-terminal SAP domain of Ku70 facilitates the initial phases of the Ku-DNA interaction but does not affect the sliding process. This suggests that the SAP domain may either establish the first interactions with DNA, or stabilize these initial interactions during loading. To assess the biological role of the SAP domain, we generated Arabidopsis plants expressing Ku lacking the SAP domain. Intriguingly, despite the decreased efficiency of the ΔSAP Ku complex in loading onto DNA, the mutant plants exhibited full proficiency in classical NHEJ and telomere maintenance. This indicates that the speed with which Ku loads onto telomeres or DNA double-strand breaks is not the decisive factor in stabilizing these DNA structures.

• MeSH
• antigen Ku * genetika   metabolismus   MeSH
• DNA genetika   metabolismus   MeSH
• kinetika MeSH
• oprava DNA spojením konců MeSH
• oprava DNA * MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antigen Ku * MeSH
• DNA MeSH

As a link between a stable genome and a dynamic environment, epigenetics is a promising tool for mapping age-related changes in human DNA. Methylated cytosine changes at specific loci are generally less studied in sperm DNA than in somatic cell DNA. Age-related methylation changes can be connected to various reproductive health problems and multiple disorders in offspring. In addition, they can be helpful in forensic fields, where testing of specific loci in semen samples found at sexual assault crime scenes can predict a perpetrator's age and narrow down the police investigation. This review focuses on age-related methylation changes in sperm. It covers the biological role of methylation, methylation testing techniques and the implications of methylation changes in forensics and clinical practice.

DNA methylation is a biological process that can change the activity of a gene without changing its sequence. We do not know much about DNA methylation in sperm and what changes methylation undergoes during the lifespan. These changes can, however, be important both for health and solving crimes. Presperm cells renew themselves, which gives rise to new sperm cells, from youth to death, accumulating cell divisions prone to error. This is why sperm cells are affected by age more than nondividing eggs. Methylation is specific in different tissues of the body. The ratio between number of sperm cells, white blood cells, and other cell types is highly variable and hardly predicted, which may distort the results. Clinical studies have shown that older fathers have worse reproductive health. Their children can develop metabolic, neurological and behavioral disorders. This also applies to younger men whose DNA methylation pattern is similar to that of older men. Methylation changes allow us to build a model capable of predicting the age of an unknown person with a mean error of about 5 years. This can be helpful for police investigators in cases of sexual assault, when biological material is found but there is no match in the police database.

• Klíčová slova
• age prediction, epigenome, forensic phenotyping, methylation, semen, sperm,
• MeSH
• DNA metabolismus   MeSH
• epigeneze genetická MeSH
• lidé MeSH
• metylace DNA * MeSH
• sperma * MeSH
• spermie metabolismus   MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Názvy látek
• DNA MeSH