Biophysics is an interdisciplinary science that applies the theories and methods of physics to understand biological systems. It encompasses a wide range of topics, from the molecular mechanisms within cells to the physical properties of organisms and ecosystems. The goal of biophysics is to uncover the physical principles underlying the structure and function of biological molecules, cells, and cellular systems, providing a deeper understanding of life itself. The Institute of Biophysics, Czech Academy of Sciences (IBP) stands as a beacon of excellence in the field of biophysical research in the Czech Republic. This article delves into its history, structure, research areas, and major scientific achievements, highlighting the role of IBP in the global scientific community.
- Klíčová slova
- Biophysical methods, Biophysics, DNA damage repair, Ionizing radiation, Radiotherapy,
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
N4-acetylcytidine (ac4C) is a post-transcriptional RNA modification that plays a crucial role in the epitranscriptome, influencing gene expression and cellular function. This modification occurs at the cytosine base, where an acetyl group is installed to the nitrogen at the 4th position (N4). This co-transcription modification affects RNA stability, RNA structure, and translation efficiency. Recent studies have uncovered a potential link between RNA modifications and DNA repair mechanisms, suggesting that ac4C-modified or methylated RNAs may interact with factors involved in DNA repair pathways; thus, influencing the cellular response to DNA damage. Dysregulation of modified RNAs, including ac4C RNA, has been implicated in cancer development, where aberrant levels of these RNAs may contribute to oncogenic transformation by altering genome stability and the expression of key genes regulating cell proliferation, cell cycle progression, and apoptosis. Understanding the dynamics of modified RNAs offers promising insights into the role of epitranscriptome in DNA repair processes and cancer treatment.
- Klíčová slova
- DNA damage repair, N-acetylcytidine, NAT10, RNA modifications, epigenetics, epitranscriptomics,
- MeSH
- cytidin * analogy a deriváty metabolismus MeSH
- epigeneze genetická * MeSH
- lidé MeSH
- nádory * genetika metabolismus MeSH
- oprava DNA * MeSH
- posttranskripční úpravy RNA * MeSH
- regulace genové exprese u nádorů MeSH
- RNA * metabolismus genetika MeSH
- transkriptom * MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
- Názvy látek
- cytidin * MeSH
- N-acetylcytidine MeSH Prohlížeč
- RNA * MeSH
PURPOSE: Parasites of genus Encephalitozoon are well known pathogens of domestic animals however less attention was paid to its spread among wildlife that can play an important role of reservoir of infection. The aim of the study was to conduct molecular detection and genotype characterization of Encephalitozoon spp. in wild small mammals trapped in localities both near to and at a large distance from residential areas. METHODS: In total, 300 wild small mammals (274 Rodentia and 26 Eulipotyphla) were trapped in 41 localities of the Czech Republic and tested by nested PCR for Encephalitozoon spp. RESULTS: The DNA of Encephalitozoon spp. was proved in tissues (brain or liver) of 11% (32/300) of animals. There was a statistically significant difference (p < 0.001) in positivity among animal species with the most infected species Micromys minutus (50%, 4/8) and Myodes glareolus (17%, 9/53). There was also statistically significant difference (p < 0.001) between localities with the higher positivity (29%, 12/42) in localities near to residential areas, compared to localities with a large distance from residential areas (8%, 20/258). Sex and age of wild small mammals did not have effect on their positivity. Genotyping analysis revealed E. cuniculi genotype II in 22 samples and E. hellem genotype 1 A in one sample. CONCLUSION: This study brings new information on the molecular characterization of Encephalitozoon spp. isolated from wild small mammals trapped in two different areas (localities in near to residential areas and localities with a large distance from residential areas).
- Klíčová slova
- Encephalitozoonosis, Genotyping, Urban area, Wildlife, Zoonosis,
- MeSH
- divoká zvířata * parazitologie MeSH
- Encephalitozoon cuniculi genetika izolace a purifikace klasifikace MeSH
- Encephalitozoon * genetika izolace a purifikace klasifikace MeSH
- encephalitozoonóza * veterinární epidemiologie parazitologie mikrobiologie MeSH
- genotyp * MeSH
- hlodavci * parazitologie MeSH
- hmyzožravci parazitologie MeSH
- játra parazitologie MeSH
- mozek parazitologie MeSH
- polymerázová řetězová reakce MeSH
- zdroje nemoci * parazitologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Česká republika MeSH
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.
- Klíčová slova
- CRISPR-dCas9, DNA methylation, RNA modifications, epigenetics, gene therapy, histone code,
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
Bats may carry various viruses and bacteria which can be harmful to humans, but little is known about their role as a parasitic source with zoonotic potential. The aim of this study was to test wild bats for the presence of selected parasites: Toxoplasma gondii, Neospora caninum and microsporidia Encephalitozoon spp. In total, brain and small intestine tissues of 100 bats (52 Myotis myotis, 43 Nyctalus noctula and 5 Vespertilio murinus) were used for the DNA isolation and PCR detection of the abovementioned agents. Toxoplasma gondii DNA was detected by real-time PCR in 1% of bats (in one male of M. myotis), while all bats were negative for N. caninum DNA. Encephalitozoon spp. DNA was detected by nested PCR in 25% of bats, including three species (twenty-two M. myotis, two N. noctula and one V. murinus). Positive samples were sequenced and showed homology with the genotypes Encephalitozoon cuniculi II and Encephalitozoon hellem 2C. This is the first study on wild vespertilionid bats from Central Europe and worldwide, with a relatively high positivity of Encephalitozoon spp. detected in bats.
- Klíčová slova
- insectivore, microsporidiosis, molecular methods, neosporosis, toxoplasmosis,
- MeSH
- Chiroptera * MeSH
- Encephalitozoon * genetika MeSH
- kokcidióza * veterinární MeSH
- kvantitativní polymerázová řetězová reakce MeSH
- lidé MeSH
- Neospora * genetika MeSH
- paraziti * genetika MeSH
- Toxoplasma * genetika MeSH
- toxoplazmóza zvířat * parazitologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Evropa 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.
- Klíčová slova
- ACE2, embryonic heart, human kidney embryonic cells, lung cancer cells, renin,
- 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
- Názvy látek
- angiotensin-konvertující enzym 2 MeSH
- renin 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).
- Klíčová slova
- Apodemus, Myodes, Q fever, Serology, Tularemia,
- MeSH
- Arvicolinae MeSH
- Coxiella burnetii izolace a purifikace MeSH
- Francisella tularensis izolace a purifikace MeSH
- koinfekce epidemiologie mikrobiologie veterinární MeSH
- krtci MeSH
- Murinae 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 epidemiologie MeSH
- Názvy látek
- protilátky bakteriální 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.
- Klíčová slova
- 53BP1, BRCA1, DNA damage, epigenetics, histone modifications,
- 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
- Názvy látek
- 53BP1 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.
- Klíčová slova
- LAT, PCR, Serology, Stray cats, Toxoplasmosis,
- 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
- Názvy látek
- protilátky protozoální MeSH
- protozoální DNA 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.
- Klíčová slova
- DNA damage, Lap2α, confocal microscopy, epigenetics, immunohistochemistry, lamins,
- MeSH
- 53BP1 genetika metabolismus MeSH
- červený fluorescenční protein 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
- Názvy látek
- 53BP1 MeSH
- chromatin MeSH
- DNA vazebné proteiny MeSH
- gamma-H2AX protein, mouse MeSH Prohlížeč
- histony MeSH
- lamin typ A MeSH
- lamina-associated polypeptide 2 MeSH Prohlížeč
- luminescentní proteiny MeSH
- membránové proteiny MeSH
- rekombinantní fúzní proteiny MeSH
- Trp53bp1 protein, mouse MeSH Prohlížeč
