The aim of this study was to assess two protocols for their capacities to simultaneously isolate RNA, mtDNA and ncDNA from mammalian cells. We compared the Invitrogen TRIzol-based method and Qiagen DNeasy columns, using the HepG2 cell line and human primary glioblastoma stem cells. Both methods allowed the isolation of all three types of nucleic acids and provided similar yields in mtDNA. However, the yield in ncDNA was more than tenfold higher on columns, as observed for both cell types. Conversely, the TRIzol method proved more reproducible and was the method of choice for isolating RNA from glioblastoma cells, as demonstrated for the housekeeping genes RPLP0 and RPS9.

• MeSH
• biochemie metody   MeSH
• buněčné jádro metabolismus   MeSH
• buňky Hep G2 MeSH
• glioblastom metabolismus   patologie   MeSH
• lidé MeSH
• messenger RNA izolace a purifikace   MeSH
• mitochondriální DNA izolace a purifikace   MeSH
• nádorové kmenové buňky metabolismus   MeSH
• reagenční diagnostické soupravy MeSH
• RNA izolace a purifikace   MeSH
• savci metabolismus   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The mitochondrial DNA of diplonemid and kinetoplastid protists is known for its suite of bizarre features, including the presence of concatenated circular molecules, extensive trans-splicing and various forms of RNA editing. Here we report on the existence of another remarkable characteristic: hyper-inflated DNA content. We estimated the total amount of mitochondrial DNA in four kinetoplastid species (Trypanosoma brucei, Trypanoplasma borreli, Cryptobia helicis, and Perkinsela sp.) and the diplonemid Diplonema papillatum. Staining with 4',6-diamidino-2-phenylindole and RedDot1 followed by color deconvolution and quantification revealed massive inflation in the total amount of DNA in their organelles. This was further confirmed by electron microscopy. The most extreme case is the ∼260 Mbp of DNA in the mitochondrion of Diplonema, which greatly exceeds that in its nucleus; this is, to our knowledge, the largest amount of DNA described in any organelle. Perkinsela sp. has a total mitochondrial DNA content ~6.6× greater than its nuclear genome. This mass of DNA occupies most of the volume of the Perkinsela cell, despite the fact that it contains only six protein-coding genes. Why so much DNA? We propose that these bloated mitochondrial DNAs accumulated by a ratchet-like process. Despite their excessive nature, the synthesis and maintenance of these mtDNAs must incur a relatively low cost, considering that diplonemids are one of the most ubiquitous and speciose protist groups in the ocean. © 2018 IUBMB Life, 70(12):1267-1274, 2018.

• MeSH
• Euglenozoa genetika   MeSH
• fylogeneze MeSH
• Kinetoplastida genetika   MeSH
• mitochondriální DNA genetika   izolace a purifikace   ultrastruktura   MeSH
• mitochondrie genetika   MeSH
• trans-splicing genetika   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The Ethiopian highlands are the most extensive complex of mountainous habitats in Africa. The presence of the Great Rift Valley (GRV) and the striking elevational ecological gradients inhabited by recently radiated Ethiopian endemics, provide a wide spectrum of model situations for evolutionary studies. The extant species of endemic rodents, often markedly phenotypically differentiated, are expected to possess complex genetic features which evolved asa consequence of the interplay between geomorphology and past climatic changes. In this study, we used the largest available multi-locus genetic dataset of the murid genus Stenocephalemys (347 specimens from ca 40 localities across the known distributional area of all taxa) to investigate the relative importance of disruptive selection, temporary geographic isolation and introgression in their adaptive radiations in the Pleistocene. We confirmed the four main highly supported mitochondrial (mtDNA) clades that were proposed as four species in a previous pilot study: S. albipes is a sister species of S. griseicauda (both lineages are present on both sides of the GRV), while the second clade is formed by two Afro-alpine species, S. albocaudata (east of GRV) and the undescribed Stenocephalemys sp. A (west of GRV). There is a clear elevational gradient in the distribution of the Stenocephalemys taxa with two to three species present at different elevations of the same mountain range. Surprisingly, the nuclear species tree corresponded only a little to the mtDNA tree. Multispecies coalescent models based on six nuclear markers revealed the presence of six separate gene pools (i.e. candidate species), with different topology. Phylogenetic analysis, together with the geographic distribution of the genetic groups, suggests a complex reticulate evolution. We propose a scenario that involves (besides classical allopatric speciation) two cases of disruptive selection along the elevational ecological gradient, multiple crosses of GRV in dry and cold periods of the Pleistocene, followed by hybridization and mtDNA introgression on imperfect reproductive barriers. Spatial expansion of the currently most widespread "albipes" mtDNA clade was followed by population fragmentation, lineage sorting and again by hybridization and mtDNA introgression. Comparison of this genetic structure to other Ethiopian endemic taxa highlight the geographical areas of special conservation concern, where more detailed biodiversity studies should be carried out to prevent many endemic taxa from going extinct even before they are recognized.

• MeSH
• cytochromy b chemie   klasifikace   genetika   MeSH
• ekosystém MeSH
• fylogeneze MeSH
• haplotypy MeSH
• hybridizace genetická MeSH
• karyotyp MeSH
• mitochondriální DNA chemie   izolace a purifikace   metabolismus   MeSH
• molekulární evoluce * MeSH
• Murinae anatomie a histologie   klasifikace   genetika   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Etiopie MeSH

The isolation of populations in the Iberian, Italian and Balkan peninsulas during the ice ages define four main paradigms that explain much of the known distribution of intraspecific genetic diversity in Europe. In this study we investigated the phylogeography of a wide-spread bat species, the bent-winged bat, Miniopterus schreibersii around the Mediterranean basin and in the Caucasus. Environmental Niche Modeling (ENM) analysis was applied to predict both the current distribution of the species and its distribution during the last glacial maximum (LGM). The combination of genetics and ENM results suggest that the populations of M. schreibersii in Europe, the Caucasus and Anatolia went extinct during the LGM, and the refugium for the species was a relatively small area to the east of the Levantine Sea, corresponding to the Mediterranean coasts of present-day Syria, Lebanon, Israel, and northeastern and northwestern Egypt. Subsequently the species first repopulated Anatolia, diversified there, and afterwards expanded into the Caucasus, continental Europe and North Africa after the end of the LGM. The fossil record in Iberia and the ENM results indicate continuous presence of Miniopterus in this peninsula that most probably was related to the Maghrebian lineage during the LGM, which did not persist afterwards. Using our results combined with similar findings in previous studies, we propose a new paradigm explaining the general distribution of genetic diversity in Europe involving the recolonization of the continent, with the main contribution from refugial populations in Anatolia and the Middle East. The study shows how genetics and ENM approaches can complement each other in providing a more detailed picture of intraspecific evolution.

• MeSH
• biologické modely MeSH
• Chiroptera klasifikace   genetika   MeSH
• fylogeneze MeSH
• fylogeografie MeSH
• genetická variace MeSH
• mitochondriální DNA izolace a purifikace   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Balkánský poloostrov MeSH
• Evropa MeSH
• severní Afrika MeSH
• Střední východ MeSH

Introduction and Objectives: One of the characteristic changes of tumor formation is accumulation of genetic disorders in mitochondrial and nuclear genome. Mitochondrial disorders, from its side, are responsible for failure of metabolism, apoptosis, cell growth, formation of reactive oxygen species, etc. Overprpoduction of reactive oxygen species (ROS) significantly impacts the respiration chain enzymes and entirely the antioxidant system of mitochondria. Finally this may become a favorable condition for normal cells transformation. The purpose of the presented work was to study the mitochondrial defects and to establish their role in prostate cancer development. Results: Experimental results demonstrate significant increase of the activity of mitochondrial succinate dehydrogenaze (complex II) of the malignant epithelial cells of prostate, and slight changes in cytochrome oxydase (complex IV) activity. Also significant activation of the antioxidant system (glutathione-dependant system) of mitochondria in prostate malignant epithelial cells was revealed. Conclusion: The above mentioned mitochondrial changes (II and IV complexes of respiration chain, activity of the antioxidant system) partially demonstrate the alterations in mitochondrial energy metabolism, which from its side, may indicate to resistance of prostate cancer cells and correspondingly to intensification of proliferation processes.

• MeSH
• citrátový cyklus MeSH
• glutathion metabolismus   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mitochondriální DNA * genetika   metabolismus   genetika   izolace a purifikace   metabolismus   MeSH
• nádory prostaty * genetika   MeSH
• přenos energie MeSH
• reaktivní formy kyslíku * MeSH
• respirační komplex IV MeSH
• senioři MeSH
• sukcinátdehydrogenasa * diagnostické užití   MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH

Parkinsonova choroba patrí medzi najčastejšie sa vyskytujúce neurodegeneratívne ochorenie so závažnými socioekonomickými dôsledkami. Z hľadiska súčasných poznatkov ide o ochorenie nevyliečiteľné, avšak včasnou diagnostikou a vhodne zahájenou liečbou sa dá prispieť k zlepšeniu kvality života pacienta. Príčina vzniku Parkinsonovej choroby nie je dodnes presne známa. V súčasnosti sa množia dôkazy podporujúce teóriu, že oxidatívny stres a mitochondriálna dysfunkcia môžu mať zásadný význam v patogenéze ochorenia. Výskum mechanizmov zahrnutých v zložitom procese neurodogenerácie PCh by mohla priniesť možnosť identifikácie rizikových a presymptomatických jedincov, ako aj identifikovanie terapeutického cieľa a následnej neuroprotektívnej liečby.

Parkinson's disease is the most common neurodegenerative disorder, with serious socio-economic consequences. At present the disorder appears incurable, but well-timed diagnostics and appropriate therapy can improve the quality of life for those who suffer from it. Recent research appears to support a hypothesis that oxidative damage and mitochondrial dysfunction may play a primary role in the pathogenesis of PD. Further investigation of the mechanisms implicated in the complex process of neurodegeneration may result in the possibility of identifying individuals at risk and presymptomatic patients, and also determining proper therapeutic targets and subsequent neuroprotective treatments.

• MeSH
• apoptóza genetika   imunologie   MeSH
• dopamin chemie   metabolismus   škodlivé účinky   MeSH
• kyselina močová krev   metabolismus   MeSH
• lidé MeSH
• mitochondriální DNA izolace a purifikace   MeSH
• mitochondriální nemoci diagnóza   etiologie   genetika   MeSH
• mutace fyziologie   genetika   imunologie   MeSH
• neurodegenerativní nemoci etiologie   farmakoterapie   MeSH
• neurony chemie   metabolismus   účinky léků   MeSH
• neuroprotektivní látky metabolismus   terapeutické užití   MeSH
• oxidační stres fyziologie   genetika   imunologie   MeSH
• Parkinsonova nemoc diagnóza   etiologie   farmakoterapie   MeSH
• poruchy metabolismu vápníku etiologie   genetika   MeSH
• poruchy metabolismu železa etiologie   genetika   MeSH
• stárnutí fyziologie   genetika   imunologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• přehledy MeSH

U 18letého mladíka se během jednoho týdne významně zhoršilo vidění nejdříve levým a po dvou měsících i pravým okem bez periokulární bolesti. Ostatní neurologický nález byl v normě. Na magnetické rezonanci (MR) mozku byl normální obraz, v optických nervech a chiazmatu se nalezla tečkovitá hypersignální ložiska. V likvoru byly opakovaně přítomny oligoklonální pásy IgG. Proto bylo onemocnění považováno za roztroušenou sklerózu. Během jednoho roku sledování se zrak nezlepšil a neobjevilo se žádné další postižení. Výsledky pomocných vyšetření byly v rozporu s atypickým klinickým průběhem a normálním MR obrazem. tento nesoulad nás vedl k podezření na jiné diagnózy. antiakvaporinové protilátky nepodpořily možnost Devicovy optické neuromyelitidy. Nakonec se prokázala mutace v mitochondriální DNa svědčící pro Leberovu hereditární optickou neuropatii (LHON). Ke správnému určení vzácné diagnózy u složitého pacienta přispěla široká mezioborová spolupráce včetně zahraniční laboratoře. Případ dokládá zavádějící úlohu oligoklonálních pásů v likvoru a má směřovat k většímu povědomí o LHON v odborné veřejnosti

Vision deteriorated in an 18-year-old man without periocular pain during one week in the left eye first and in the right eye two months later. Neurological findings were otherwise normal. Magnetic resonance imaging (MRI) of the brain was normal and there were high signals within optic nerves and chiasma. Oligoclonal IgG bands were repeatedly detected in the cerebrospinal fluid. As a result, the disorder was considered to be multiple sclerosis. The vision failed to improve during one-year follow-up and no other impairment appeared. the results of auxiliary investigations were not in agreement with the atypical clinical course of the disease and normal MRI of the brain. We considered other diagnoses due to this discrepancy. anti-aquaporin antibodies did not support neuromyelitis optica (Devic disease). Finally, the mitochondrial DNA mutation confirmed Leber hereditary optic neuropathy (LHON). the correct diagnosis was achieved after broad interdisciplinary cooperation including foreign laboratory. This case illustrates the misleading role of oligoclonal bands in cerebrospinal fluid and should increase general knowledge about LHON in physicians.

• MeSH
• diferenciální diagnóza MeSH
• financování organizované MeSH
• Leberova atrofie zrakového nervu diagnóza   etiologie   patofyziologie   MeSH
• lidé MeSH
• magnetická rezonanční tomografie využití   MeSH
• mezioborová komunikace MeSH
• mitochondriální DNA genetika   izolace a purifikace   MeSH
• mladiství MeSH
• mutace genetika   MeSH
• oligoklonální proužky izolace a purifikace   MeSH
• roztroušená skleróza diagnóza   MeSH
• zrakové evokované potenciály genetika   MeSH
• Check Tag
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• Publikační typ
• kazuistiky MeSH

Despite the large size of the contemporary nomadic Fulani population (roughly 13 million people), the genetic diversity and degree of differentiation of Fulanis compared to other sub-Saharan populations remain unknown. We sampled four Fulani nomad populations (n = 186) in three countries of sub-Saharan Africa (Chad, Cameroon, and Burkina Faso) and analyzed sequences of the first hypervariable segment of the mitochondrial DNA. Most of the haplotypes belong to haplogroups of West African origin, such as L1b, L3b, L3d, L2b, L2c, and L2d (79.6% in total), which are all well represented in each of the four geographically separated samples. The haplogroups of Western Eurasian origin, such as J1b, U5, H, and V, were also detected but in rather low frequencies (8.1% in total). As in African hunter-gatherers (Pygmies and Khoisan) and some populations from central Tunisia (Kesra and Zriba), three of the Fulani nomad samples do not reveal significant negative values of Fu's selective neutrality test. The multidimensional scaling of FST genetic distances of related sub-Saharan populations and the analysis of molecular variance (AMOVA) show clear and close relationships between all pairs of the four Fulani nomad samples, irrespective of their geographic origin. The only group of nomadic Fulani that manifests some similarities with geographically related agricultural populations (from Guinea-Bissau and Nigeria) comes from Tcheboua in northern Cameroon.

• MeSH
• financování organizované MeSH
• haplotypy genetika   MeSH
• lidé MeSH
• mitochondriální DNA genetika   izolace a purifikace   MeSH
• osoby s přechodným pobytem a migranti MeSH
• populační genetika metody   MeSH
• Check Tag
• lidé MeSH
• Geografické názvy
• subsaharská Afrika MeSH

• MeSH
• diabetes mellitus 2. typu genetika   komplikace   patofyziologie   MeSH
• hluchota genetika   komplikace   patofyziologie   MeSH
• lidé MeSH
• mitochondriální DNA genetika   chemie   izolace a purifikace   MeSH
• mitochondriální encefalomyopatie diagnóza   etiologie   patofyziologie   MeSH
• RNA transferová Leu biosyntéza   genetika   MeSH
• sukcinát: cytochrom c oxidoreduktasa MeSH
• Check Tag
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• kazuistiky MeSH
• srovnávací studie MeSH

• MeSH
• Candida genetika   metabolismus   MeSH
• DNA vazebné proteiny izolace a purifikace   metabolismus   MeSH
• elektroforéza v polyakrylamidovém gelu MeSH
• mitochondriální DNA chemie   izolace a purifikace   metabolismus   MeSH
• oligodeoxyribonukleotidy chemie   metabolismus   MeSH
• substrátová specifita MeSH
• telomery MeSH