Gene therapy is a focus of interest in both human and veterinary medicine, especially in recent years due to the potential applications of CRISPR/Cas9 technology. Another relatively new approach is that of epigenetic therapy, which involves an intervention based on epigenetic marks, including DNA methylation, histone post-translational modifications, and post-transcription modifications of distinct RNAs. The epigenome results from enzymatic reactions, which regulate gene expression without altering DNA sequences. In contrast to conventional CRISP/Cas9 techniques, the recently established methodology of epigenetic editing mediated by the CRISPR/dCas9 system is designed to target specific genes without causing DNA breaks. Both natural epigenetic processes and epigenetic editing regulate gene expression and thereby contribute to maintaining the balance between physiological functions and pathophysiological states. From this perspective, knowledge of specific epigenetic marks has immense potential in both human and veterinary medicine. For instance, the use of epigenetic drugs (chemical compounds with therapeutic potential affecting the epigenome) seems to be promising for the treatment of cancer, metabolic, and infectious diseases. Also, there is evidence that an epigenetic diet (nutrition-like factors affecting epigenome) should be considered as part of a healthy lifestyle and could contribute to the prevention of pathophysiological processes. In summary, epigenetic-based approaches in human and veterinary medicine have increasing significance in targeting aberrant gene expression associated with various diseases. In this case, CRISPR/dCas9, epigenetic targeting, and some epigenetic nutrition factors could contribute to reversing an abnormal epigenetic landscape to a healthy physiological state.
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
Toxoplasmosis is a globally spread disease, affecting humans and many animal species, including birds. Antibodies to Toxoplasma gondii were detected in ostriches from South and North America, Africa and Asia. Except for one study from Spain, there is a lack of information about T. gondii seroprevalence in ostriches from Europe. For this reason, the aim of the study was to detect antibodies to T. gondii in farm-reared ostriches from the Czech Republic. Serum samples of 409 ostriches (Struthio camelus), collected at 9 farms were tested by Latex agglutination test. Antibodies to T. gondii were detected in 149 (36%) birds with a statistical difference for individual farms (8%-71%, p = 0.0121), and regions (8%-65%, p = 0.002). Seropositivity did not statistically differ (p > 0.05) in size of farms (50% and 35% on small and large farms, respectively), sex of birds (38% and 35% in males and females, respectively), season and year of collection. Tissue samples (brain, heart, and pectoral muscle) of 105 birds were also tested by PCR to detect T. gondii DNA. The parasite T. gondii was detected in the brain and heart of one seronegative ostrich (1%) from a small farm. Based on our results, we can assume that ostriches may present high risk of toxoplasmosis for humans through consumption of raw or undercooked ostrich meat and even seronegative individuals could harbor T. gondii in their tissues. To our knowledge, this is the first serological detection of T. gondii in ostriches in the Czech Republic, and the first PCR detection in Europe.
- MeSH
- farmy MeSH
- lidé MeSH
- protilátky protozoální krev MeSH
- protozoální DNA analýza MeSH
- rizikové faktory MeSH
- Struthioniformes * krev parazitologie MeSH
- Toxoplasma * genetika imunologie MeSH
- toxoplazmóza zvířat * krev diagnóza epidemiologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Česká republika MeSH
Wild rodents are an important source of the tick-borne pathogens Coxiella burnetii and Francisella tularensis. The aim of our study was to assess the prevalence of antibodies and possible coexistence of these pathogens in wild small mammals from three localities in the Czech Republic. A total of 614 wild small mammals (324 Apodemus flavicollis, 145 Myodes glareolus, 50 Sorex araneus, 48 A. sylvaticus, 40 A. agrarius, six Microtus arvalis and one Talpa europaea) were trapped between 2012 and 2015. Their sera or heart extracts were examined by modified indirect enzyme-linked immunosorbent assay, with the detection of antibodies against C. burnetii and F. tularensis in 12 % and 7 % of animals, respectively; coinfection was identified in 4.4 % of animals. The prevalence of C. burnetii and F. tularensis antibodies statistically differed according to animal species and sex (p < 0.05); the seroprevalence of C. burnetii (p < 0.05) also differed in the sampling period. The highest prevalence of antibodies against C. burnetii and F. tularensis was detected in the case of M. glareolus (24 % and 14 %, respectively).
- MeSH
- Arvicolinae MeSH
- Coxiella burnetii izolace a purifikace MeSH
- Francisella tularensis izolace a purifikace MeSH
- koinfekce epidemiologie mikrobiologie veterinární MeSH
- krtci MeSH
- Murinae MeSH
- nemoci hlodavců epidemiologie mikrobiologie MeSH
- prevalence MeSH
- protilátky bakteriální krev MeSH
- Q-horečka epidemiologie mikrobiologie veterinární MeSH
- rejskovití MeSH
- séroepidemiologické studie MeSH
- tularemie epidemiologie mikrobiologie veterinární MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika MeSH
ACE2 was observed as the cell surface receptor of the SARS-CoV-2 virus. Interestingly, we also found ACE2 positivity inside the cell nucleus. The ACE2 levels changed during cell differentiation and aging and varied in distinct cell types. We observed ACE2 depletion in the aortas of aging female mice, similarly, the aging caused ACE2 decrease in the kidneys. Compared with that in the heart, brain and kidneys, the ACE2 level was the lowest in the mouse lungs. In mice exposed to nicotine, ACE2 was not changed in olfactory bulbs but in the lungs, ACE2 was upregulated in females and downregulated in males. These observations indicate the distinct gender-dependent properties of ACE2. Differentiation into enterocytes, and cardiomyocytes, caused ACE2 depletion. The cardiomyogenesis was accompanied by renin upregulation, delayed in HDAC1-depleted cells. In contrast, vitamin D2 decreased the renin level while ACE2 was upregulated. Together, the ACE2 level is high in non-differentiated cells. This protein is more abundant in the tissues of mouse embryos and young mice in comparison with older animals. Mostly, downregulation of ACE2 is accompanied by renin upregulation. Thus, the pathophysiology of COVID-19 disease should be further studied not only by considering the ACE2 level but also the whole renin-angiotensin system.
- MeSH
- angiotensin-konvertující enzym 2 metabolismus MeSH
- buněčná diferenciace fyziologie MeSH
- buňky A549 MeSH
- buňky HT-29 MeSH
- COVID-19 epidemiologie patologie virologie MeSH
- HEK293 buňky MeSH
- lidé MeSH
- myši MeSH
- pandemie MeSH
- regulace genové exprese fyziologie MeSH
- renin-angiotensin systém fyziologie MeSH
- renin metabolismus MeSH
- SARS-CoV-2 patogenita MeSH
- sexuální faktory MeSH
- stárnutí fyziologie MeSH
- věkové faktory MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Nuclear architecture plays a significant role in DNA repair mechanisms. It is evident that proteins involved in DNA repair are compartmentalized in not only spontaneously occurring DNA lesions or ionizing radiation-induced foci (IRIF), but a specific clustering of these proteins can also be observed within the whole cell nucleus. For example, 53BP1-positive and BRCA1-positive DNA repair foci decorate chromocenters and can appear close to nuclear speckles. Both 53BP1 and BRCA1 are well-described factors that play an essential role in double-strand break (DSB) repair. These proteins are members of two protein complexes: 53BP1-RIF1-PTIP and BRCA1-CtIP, which make a "decision" determining whether canonical nonhomologous end joining (NHEJ) or homology-directed repair (HDR) is activated. It is generally accepted that 53BP1 mediates the NHEJ mechanism, while HDR is activated via a BRCA1-dependent signaling pathway. Interestingly, the 53BP1 protein appears relatively quickly at DSB sites, while BRCA1 is functional at later stages of DNA repair, as soon as the Mre11-Rad50-Nbs1 complex is recruited to the DNA lesions. A function of the 53BP1 protein is also linked to a specific histone signature, including phosphorylation of histone H2AX (γH2AX) or methylation of histone H4 at the lysine 20 position (H4K20me); therefore, we also discuss an epigenetic landscape of 53BP1-positive DNA lesions.
- MeSH
- 53BP1 genetika metabolismus MeSH
- buněčné jádro genetika metabolismus MeSH
- fosforylace MeSH
- lidé MeSH
- oprava DNA * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
Histone posttranslational modifications regulate diverse nuclear functions, including DNA repair. Here, we use mass spectrometry, western blotting, immunohistochemistry and advanced confocal microscopy in order to show radiation-specific changes in the histone signature. We studied wild-type mouse embryonic stem cells (mESCs) and mESCs with a depletion of histone deacetylase 1 (HDAC1), which plays a role in DNA repair. Irradiation by γ-rays increased the S139 phosphorylation of histone H2AX but reduced the level of the H3K9-R17 peptide, which contains S10 phosphorylation (H3S10ph). On an individual cellular level, H3S10ph was low in highly γH2AX-positive UV laser-induced DNA lesions, and this nuclear distribution pattern was not changed by HDAC1 depletion. Despite this fact, spontaneous γH2AX-positive DNA lesions colocalized with large H3S10ph-positive nuclear bodies that appear in the G2 phase of the cell cycle. Similarly, by FLIM-FRET analysis, we observed an interaction between H3S10ph and γH2AX in the G2 phase. However, this interaction was reduced when cells were exposed to γ-rays. A mutual link between H3S10ph and γH2AX was not observed in the G1 phase of the cell cycle. Together, our data show that despite the fact that H3S10ph is not directly involved in DNA repair, a decrease in H3S10 phosphorylation and weakened interaction between H3S10ph and γH2AX is a result of radiation-induced damage of the genome. In this case, γ-irradiation also decreased the number of cells in the G1 phase, characterized by no interaction between H3S10ph and γH2AX.
- MeSH
- fosforylace účinky záření MeSH
- G1 fáze účinky záření MeSH
- G2 fáze účinky záření MeSH
- HeLa buňky MeSH
- histony genetika metabolismus MeSH
- lidé MeSH
- myši MeSH
- záření gama škodlivé účinky MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Toxoplasma gondii is protozoan parasite with ability of causing disease in wide-spectrum of animals; many species of animals in captivity died of clinical toxoplasmosis. The monitoring of T. gondii antibodies in zoo animals can be an important indicator of T. gondii circulation in zoo. The aim of this study was to examine sera of animals from eight Czech zoos by latex agglutination test with statistical evaluation and detect T. gondii DNA in stray cats and rodents captured in the zoos. T. gondii antibodies were detected in 33% of 1043 zoo animals without statistical difference between birds (27%, n = 74) and mammals (33%, n = 969). In birds, the chance to be infected with T. gondii was higher in Accipitriformes (71%) compared to Pelecaniformes (6%) (p < 0.0001). In mammals, the chance to be infected with T. gondii was higher in Carnivora (63%) compared to Cetarodactyla (30%), Perissodactyla (26%), Primates (28%) and Rodentia (13%) (p < 0.0001) and higher in Felidae (70%) compared to Bovidae (28%) and Equidae (28%) (p < 0.0001). Mammals with carnivore/scavenger way of feeding were in a higher risk of T. gondii infection compared to herbivores and omnivores (p < 0.0001). T. gondii DNA was detected in tissue of one stray cat while in none of 77 rodents caught in zoo. This study is the first report on toxoplasmosis in zoos from the Czech Republic including seroepidemiology and molecular detection.
- MeSH
- Carnivora krev imunologie parazitologie MeSH
- divoká zvířata krev imunologie parazitologie MeSH
- hlodavci krev imunologie parazitologie MeSH
- kočky MeSH
- latex fixační testy MeSH
- protilátky protozoální imunologie MeSH
- protozoální DNA krev MeSH
- ptáci krev imunologie parazitologie MeSH
- rizikové faktory MeSH
- savci krev imunologie parazitologie MeSH
- séroepidemiologické studie MeSH
- Toxoplasma genetika imunologie MeSH
- toxoplazmóza zvířat krev epidemiologie imunologie MeSH
- zvířata v ZOO krev imunologie parazitologie MeSH
- zvířata MeSH
- Check Tag
- kočky MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Česká republika epidemiologie MeSH
We studied how deficiency in lamins A/C and lamina-associated polypeptide 2α (Lap2α) affects DNA repair after irradiation. A-type lamins and Lap2α were not recruited to local DNA lesions and did not accumulate to γ-irradiation-induced foci (IRIF), as it is generally observed for well-known marker of DNA lesions, 53BP1 protein. At micro-irradiated chromatin of lmna double knockout (dn) and Lap2α dn cells, 53BP1 protein levels were reduced, compared to locally irradiated wild-type counterpart. Decreased levels of 53BP1 we also observed in whole populations of lmna dn and Lap2α dn cells, irradiated by UV light. We also studied distribution pattern of 53BP1 protein in a genome outside micro-irradiated region. In Lap2α deficient cells, identical fluorescence of mCherry-tagged 53BP1 protein was found at both microirradiated region and surrounding chromatin. However, a well-known marker of double strand breaks, γH2AX, was highly abundant in the lesion-surrounding genome of Lap2α deficient cells. Described changes, induced by irradiation in Lap2α dn cells, were not accompanied by cell cycle changes. In Lap2α dn cells, we additionally performed analysis by FLIM (Fluorescence Lifetime Imaging Microscopy) that showed different dynamic behavior of mCherry-tagged 53BP1 protein pools when it was compared with wild-type (wt) fibroblasts. This analysis revealed three different fractions of mCherry-53BP1 protein. Two of them showed identical exponential decay times (τ1 and τ3), but the decay rate of τ2 and amplitudes of fluorescence decays (A1-A3) were statistically different in wt and Lap2α dn fibroblasts. Moreover, γ-irradiation weakened an interaction between A-type lamins and Lap2α. Together, our results demonstrate how depletion of Lap2α affects DNA damage response (DDR) and how chromatin compactness is changed in Lap2α deficient cells exposed to radiation.
- MeSH
- 53BP1 genetika metabolismus MeSH
- chromatin chemie účinky záření ultrastruktura MeSH
- DNA vazebné proteiny nedostatek genetika MeSH
- embryo savčí MeSH
- fibroblasty cytologie metabolismus účinky záření MeSH
- FRAP MeSH
- histony genetika metabolismus MeSH
- lamin typ A nedostatek genetika MeSH
- luminescentní proteiny genetika metabolismus MeSH
- membránové proteiny nedostatek genetika MeSH
- myši MeSH
- oprava DNA * MeSH
- poškození DNA MeSH
- regulace genové exprese MeSH
- rekombinantní fúzní proteiny genetika metabolismus MeSH
- reportérové geny MeSH
- signální transdukce MeSH
- transformované buněčné linie MeSH
- ultrafialové záření MeSH
- záření gama MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Neospora caninum and Toxoplasma gondii are the protozoan parasites with definitive hosts from order Carnivora. Due to vertical transmission, both parasites can cause abortions and neonatal mortality that lead to significant productive and economic losses in the domestic ruminants. The aim of this study was to describe N. caninum and T. gondii seroprevalence in the group of frequently farmed captive exotic ruminants (n = 184) including Bovidae (barbary sheep, bezoar goat, common eland, American bison, water buffalo, and yak) and Camelidae (bactrian camel, guanaco, llama, and alpaca). Antibodies were tested by indirect fluorescent antibody test (IFAT) and enzyme-linked immunosorbent assay (ELISA). Higher prevalence of T. gondii antibodies (31% in IFAT and 52% in ELISA) was detected compared to N. caninum (24% in IFAT and 17% in cELISA). Mixed infection was found in 18 (10%) and 22 (12%) animals by IFAT and ELISA, respectively. Higher seroprevalence of both N. caninum and T. gondii was found in Camelidae compared to Bovidae. To author knowledge, this is the first detection of T. gondii and N. caninum antibodies in common elands and bezoar goats.
- MeSH
- ELISA veterinární MeSH
- fluorescenční protilátková technika nepřímá veterinární MeSH
- kokcidióza epidemiologie parazitologie veterinární MeSH
- Neospora imunologie MeSH
- prevalence MeSH
- přežvýkavci parazitologie MeSH
- séroepidemiologické studie MeSH
- Toxoplasma imunologie MeSH
- toxoplazmóza zvířat epidemiologie parazitologie MeSH
- velbloudovití parazitologie MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Česká republika epidemiologie MeSH
We studied the histone signature of embryonic and adult brains to strengthen existing evidence of the importance of the histone code in mouse brain development. We analyzed the levels and distribution patterns of H3K9me1, H3K9me2, H3K9me3, and HP1β in both embryonic and adult brains. Western blotting showed that during mouse brain development, the levels of H3K9me1, H3K9me2, and HP1β exhibited almost identical trends, with the highest protein levels occurring at E15 stage. These trends differed from the relatively stable level of H3K9me3 at developmental stages E8, E13, E15, and E18. Compared with embryonic brains, adult brains were characterized by very low levels of H3K9me1/me2/me3 and HP1β. Manipulation of the embryonic epigenome through histone deacetylase inhibitor treatment did not affect the distribution patterns of the studied histone markers in embryonic ventricular ependyma. Similarly, Hdac3 depletion in adult animals had no effect on histone methylation in the adult hippocampus. Our results indicate that the distribution of HP1β in the embryonic mouse brain is related to that of H3K9me1/me2 but not to that of H3K9me3. The unique status of H3K9me3 in the brain was confirmed by its pronounced accumulation in the granular layer of the adult olfactory bulb. Moreover, among the studied proteins, H3K9me3 was the only posttranslational histone modification that was highly abundant at clusters of centromeric heterochromatin, called chromocenters. When we focused on the hippocampus, we found this region to be rich in H3K9me1 and H3K9me3, whereas H3K9me2 and HP1β were present at a very low level or even absent in the hippocampal blade. Taken together, these results revealed differences in the epigenome of the embryonic and adult mouse brain and showed that the adult hippocampus, the granular layer of the adult olfactory bulb, and the ventricular ependyma of the embryonic brain are colonized by specific epigenetic marks.
- MeSH
- chromozomální proteiny, nehistonové analýza metabolismus MeSH
- fluorescenční mikroskopie MeSH
- histonlysin-N-methyltransferasa metabolismus MeSH
- imunohistochemie MeSH
- mozek embryologie metabolismus MeSH
- myši inbrední C57BL MeSH
- myši knockoutované MeSH
- myši MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
